Vol. 24 (2019)
| DOI: 10.1186/s11658-018-0126-9 Volume 24 (2019) - 24:6 | |
| Title | ANTI-TUMOR EFFECTS AND MECHANISM OF GA-13315, A NOVEL GIBBERELLIN DERIVATIVE, IN HUMAN LUNG ADENOCARCINOMA: AN IN VITRO AND IN VIVO STUDY |
| Authors | Lin Xie1, Yajuan Chen2, Jingbo Chen3, Hongbin Zhang3, Yedan Liao1, Yonghong Zhou1, Ling Zhou1 and Chen Qing2* |
| Abstract | Objective: To investigate the anti-tumor effects and the mechanism of the compound 13-chlorine-3, 15-dioxy-gibberellic acid methyl ester (GA-13315) in lung adenocarcinoma in vitro and in vivo. Methods: The antiproliferative effect of GA-13313 on the A549 cell line was determined by MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyl tetrazolium bromide) assay. A xenograft model of A549 was established to evaluate the anti-tumor effect and histopathological examination was performed to assess the toxicity of GA-13315. Apoptosis was detected by TUNEL staining in tissues and flow cytometry in cells; activation of caspase-3, caspase-8 and caspase-9 was evaluated by immunohistochemical analysis; protein levels of Bcl-2-associated X protein (Bax), B-cell lymphoma-2 (Bcl-2), caspase-4, activating transcription factor 4 (ATF4), glucose-regulated protein 78 (GRP78) and growth arrest and DNA damage-inducible gene 153 (GADD153) were determined by western blotting. Mitochondrial membrane potential (MMP) was measured by the JC-1 fluorescence probe. Results: Our results showed that GA-13315 exhibited potent, dose- and time-dependent anti-proliferative activity, and the IC50 values were 37.43 ± 2.73, 28.08 ± 7.76 and 19.29 ± 7.61 μM at 24, 48, and 72 h, respectively. The xenograft experiment revealed that tumor weight and volume were significantly decreased after GA-13315 3 mg/kg and 9 mg/kg (P < 0.05) treatment, and GA-13315 had low toxicity in bone marrow, kidney and colon tissues. GA-13315 triggered remarkable apoptosis in A549 cells at the concentration of 25.6 μM and 32 μM (P < 0.05) and activated caspase-3, − 8 and − 9. Moreover, GA-13315 induced apoptosis through the mitochondrial apoptosis pathway by elevating the Bax/Bcl-2 ratio, releasing cytochrome c and activating caspase-9 in A549 cells. In the endoplasmic reticulum apoptosis pathway, the levels of caspase-4, ATF4, GRP78 and GADD153 were markedly upregulated. Conclusions: This study suggests that GA-13315 can be considered as a promising chemotherapeutic agent with anticancer activity in treatment of lung cancer in future. |
| Keywords | Gibberellin derivatives, Lung adenocarcinoma, Antitumor, Toxicity |
| Address and Contact Information |
1 Department of Medical Oncology, Third Affiliated Hospital of Kunming Medical University/ Cancer Hospital of Yunnan Province, Kunming, China. 2 School of Pharmaceutical Science and Yunnan Key Laboratory of Pharmacology for Natural Products, Cancer Hospital of Yunnan Province, Kunming Medical University, NO.1168, West Chunrong Road, Chenggong Developing Area, Kunming 650031, China. 3 Key Laboratory of Medicinal Chemistry for Natural Resource, Ministry of Education School of Chemical Science and Technology, Yunnan University, Kunming, China. * Corresponding Author: qingchenhhh@qq.com |
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| DOI: 10.1186/s11658-018-0124-y Volume 24 (2019) - 24:7 | |
| Title | siRNA AGAINST TSG101 REDUCES PROLIFERATION AND INDUCES G0/G1 ARREST IN RENAL CELL CARCINOMA – INVOLVEMENT OF c-myc, CYCLIN E1, AND CDK2 |
| Authors | Chen Xu and Junhua Zheng* |
| Abstract | Objective: The tumor susceptibility gene 101 (TSG101) is closely associated with various tumor types, but its role in the pathogenesis of renal cell carcinoma (RCC) is still unknown. This study used RNA interference to silence the expression of TSG101 in RCC cell lines and explore the role of TSG101 in RCC. Methods: Immunohistochemistry and western blot were performed to detect the expression of TSG101 in 15 paired renal tumor samples. A small interfering RNA (siRNA) targeting TSG101 was transfected into A498 and 786-O cell lines. The Cell Counting Kit-8 (CCK-8) assay and colony formation assay were used to observe the changes in cell proliferation after transfection. Flow cytometry was used to detect the effect on the cell cycle. Western blot was conducted to study the changes of related functional proteins. Results: The expression of TSG101 was higher in RCC tissues than in adjacent normal tissues. The CCK-8 assay showed that the proliferation and colony formation of the A498 and 786-O cell lines were attenuated after suppression of TSG101. Flow cytometry showed that silencing of TSG101 induced G0/G1 arrest. The western blot results revealed that the levels of cell cycle-related proteins (c-myc, cyclin E1 and cyclin-dependent kinase 2 (CDK2)) were markedly decreased in the siRNA groups. Conclusions: TSG101 promotes proliferation of RCC cells. This positive effect on tumor growth involves activation of c-myc and cyclin E1/CDK2 and their effect on cell cycle distribution. |
| Keywords | Renal cell carcinoma, TSG101, Cell proliferation, Cell cycle |
| Address and Contact Information | Department of Urology, Tenth People’s Hospital of Tongji University, Yanchang Road 301, Shanghai 200072, China * Corresponding Author: renalzjh@163.com |
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| DOI: 10.1186/s11658-018-0129-6 Volume 24 (2019) - 24:4 | |
| Title | MiR-219-5p SUPPRESSES CELL PROLIFERATION AND CELL CYCLE PROGRESSION IN ESOPHAGEAL SQUAMOUS CELL CARCINOMA BY TARGETING CCNA2 |
| Authors | Qiang Ma |
| Abstract | Background: We investigated the potential regulatory role of miR-219-5p in esophageal squamous cell carcinoma (ESCC) and looked at the underlying mechanisms in ESCC. Methods: Real-time PCR was used to determine the levels of miR-219-5p in ESCC tissues and cell lines. The effects of miR-219-5p and cyclin A2 (CCNA2) on cell proliferation and cell cycle progression were evaluated using MTT, colony formation and flow cytometry assays with ESCC cell lines EC9706 and TE-9. Bioinformatics techniques and the luciferase reporter assay were applied to validate CCNA2 as the miR-219-5p target in ESCC cells. The mRNA and protein levels of CCNA2 were measured using real-time PCR and western blotting. Results: MiR-219-5p expression was significantly lower in ESCC tissues and cells than in healthy tissues. Upregulation of miR-219-5p repressed cell proliferation and induced cell cycle arrest at the G2/M phase. CCNA2 was identified and confirmed as a direct downstream target of miR-219-5p and its expression negatively correlated with miR-219-5p profiles in ESCC tissues. Knockdown of CCNA2 potentiated the effects of miR-219-5p on cell proliferation and cell cycle distribution. Conclusions: Our results demonstrate that miR-219-5p might function as a tumor suppressor by directly targeting CCNA2 expression. It could serve as a new therapeutic target for ESCC. |
| Keywords | Esophageal squamous cell carcinoma, miR-219-5p, CCNA2, Cell proliferation, G2/M phase arrest |
| Address and Contact Information | Department of Oncology, People’s Hospital of Xintai City, No. 1329 Xinfu Road, Xintai 271200, Shandong Province, China Corresponding Author: li_master111@163.com |
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| DOI: 10.1186/s11658-019-0137-1 Volume 24 (2019) - 24:13 | |
| Title | MicroRNA-325-3p INHIBITS CELL PROLIFERATION AND INDUCES APOPTOSIS IN HEPATITIS B VIRUS-RELATED HEPATOCELLULAR CARCINOMA BY DOWN-REGULATION OF AQUAPORIN 5 |
| Authors | Zhitao Zhang2, Yanzhen Han1* , Guangxin Sun1, Xiaohong Liu1, Xiaoyan Jia1 and Xiangjun Yu1 |
| Abstract | Background: Hepatitis B virus (HBV) infection is acknowledged as the main cause of hepatocellular carcinoma (HCC). Moreover, previous studies have revealed that microRNAs (miRNAs) widely participate in regulation of various cellular processes, such as viral replication. Hence, the purpose of this study was to investigate the roles of aquaporin 5 (AQP5) and miR-325-3p in the proliferation and apoptosis of HBV-related HCC cells. Methods: AQP5 and miR-325-3p expression in both normal and HBV-HCC tissues or cells (both Huh7–1.3 and HepG2.2.15) was detected using qRT-PCR. AQP5 expression was knocked down in HBV-related Huh7–1.3 and HepG2.2.15 cells using small interfering RNA (siRNA) technology. Down-regulation was confirmed using real-time PCR and Western blot analysis. Effects of AQP5 down-regulation on the proliferation and apoptosis were assessed. Dual luciferase reporter gene assay, Western blot and qRT-PCR were employed to evaluate the effect of miR-325-3p on the luciferase activity and expression of AQP5. Moreover, miR-325-3p mimic-induced changes in cellular proliferation and apoptosis were detected through CCK-8 assay, BrdU assay, flow cytometry analysis and ELISA. Results: In this study, the expression of AQP5 was up-regulated in human HBV-HCC tissue, Huh7–1.3 and HepG2.2.15 cells. Knockdown of AQP5 significantly inhibited the proliferation and promoted apoptosis of HBV-HCC cells. Next, miR-325-3p was obviously down-regulated in HBV-HCC. In concordance with this, MiR-325-3p directly targeted AQP5, and reduced both mRNA and protein levels of AQP5, which promoted cell proliferation and suppressed cell apoptosis in HCC cells. Overexpression of miR-325-3p dramatically inhibited cell proliferation and induced cell apoptosis. Conclusions: Our findings clearly demonstrated that introduction of miR-325-3p inhibited proliferation and induced apoptosis of Huh7–1.3 and HepG2.2.15 cells by directly decreasing AQP5 expression, and that silencing AQP5 expression was essential for the pro-apoptotic effect of miR-325-3p overexpression on Huh7–1.3 and HepG2.2.15 cells. It is beneficial to gain insight into the mechanism of HBV infection and pathophysiology of HBV-related HCC. |
| Keywords | Hepatocellular carcinoma, miR-325-3p, Hepatitis B virus, AQP5, Proliferation, Apoptosis |
| Address and Contact Information |
1 General Surgery V Ward, Affiliated Hospital of Hebei Engineering University, Handan 056002, Hebei Province, People’s Republic of China 2 Clinical Laboratory, Handan Infectious Disease Hospital, Handan 056002, Hebei Province, People’s Republic of China. * Corresponding Author: hanyanzhenhebei@163.com |
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| DOI: 10.1186/s11658-019-0140-6 Volume 24 (2019) - 24:14 | |
| Title | RELIABLE REFERENCE GENES FOR EXPRESSION ANALYSIS OF PROLIFERATING AND ADIPOGENICALLY DIFFERENTIATING HUMAN ADIPOSE STROMAL CELLS |
| Authors | Claudia Krautgasser1†, Markus Mandl1†, Florian M. Hatzmann1, Petra Waldegger1, Monika Mattesich2 and Werner Zwerschke1* |
| Abstract | Background: The proliferation and adipogenic differentiation of adipose stromal cells (ASCs) are complex processes comprising major phenotypical alterations driven by up- and downregulation of hundreds of genes. Quantitative RT-PCR can be employed to measure relative changes in the expression of a gene of interest. This approach requires constitutively expressed reference genes for normalization to counteract inter-sample variations due to differences in RNA quality and quantity. Thus, a careful validation of quantitative RT-PCR reference genes is needed to accurately measure fluctuations in the expression of genes. Here, we evaluated candidate reference genes applicable for quantitative RT-PCR analysis of gene expression during proliferation and adipogenesis of human ASCs with the immunophenotype DLK1+/CD34+/CD90+/CD105+/CD45−/CD31−. Methods: We evaluated the applicability of 10 candidate reference genes (GAPDH, TBP, RPS18, EF1A, TFRC, GUSB, PSMD5, CCNA2, LMNA and MRPL19) using NormFinder, geNorm and BestKeeper software. Results: The results indicate that EF1A and MRPL19 are the most reliable reference genes for quantitative RT-PCR analysis of proliferating ASCs. PSMD5 serves as the most reliable endogenous control in adipogenesis. CCNA2 and LMNA were among the least consistent genes. Conclusions: Applying these findings for future gene expression analyses will help elucidate ASC biology. |
| Keywords | Adipose stromal cells, Adipose stem cells, Adipogenesis, Proliferation, Differentiation, Reference gene |
| Address and Contact Information |
1 Division of Cell Metabolism and Differentiation Research, Institute for Biomedical Aging Research, University of Innsbruck, Rennweg 10, A-6020 Innsbruck, Austria 2 Department of Plastic and Reconstructive Surgery, Innsbruck Medical University, Anichstraße 35, A-6020 Innsbruck, Austria. * Corresponding Author: werner.zwerschke@uibk.ac.at † Claudia Krautgasser and Markus Mandl have equal contribution |
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| DOI: 10.1186/s11658-018-0125-x Volume 24 (2019) - 24:5 | |
| Title | TETRAHYDROXY STILBENE GLUCOSIDE ALLEVIATES PALMITIC ACID-INDUCED INFLAMMATION AND APOPTOSIS IN CARDIOMYOCYTES BY REGULATING miR-129-3p/Smad3 SIGNALING |
| Authors | Yong Zou1 and Min Kong2* |
| Abstract | Objective: Tetrahydroxy stilbene glucoside (TSG) has been reported to exert a cytoprotective effect against various toxicants. However, the function and mechanism of TSG in palmitic acid (PA)-induced inflammation and apoptosis in cardiomyocytes are still unknown. The present study was designed to investigate the post-transcriptional mechanism in TSG-treated cardiomyocytes’ inflammation and apoptosis induced by PA. Methods: The mRNA and protein levels were assayed by reverse transcription- quantitative polymerase chain reaction (RT-qPCR) and western blotting, respectively. The targeted genes were predicted by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. Cell proliferation was analyzed by CCK-8 assay. Annexin V-fluorescein isothiocyanate/polyimide (annexin V-FITC/PI) staining was used to evaluate apoptosis using flow cytometry. Results: TSG restricted the detrimental effects, including the activated inflammatory response and apoptosis, of PA in cardiomyocytes, as well as the up-regulation of miR-129-3p and down-regulation of p-Smad3 expression. In addition, bioinformatics and experimental analysis suggested that Smad3 was a direct target of miR-129-3p, which could inhibit or enhance the expression of p-Smad by transfection with miR-129-3p mimics or inhibitors, respectively. Furthermore, our results demonstrated that overexpression of Smad3 reversed the inhibition of inflammation and apoptosis by overexpression of miR-129-3p in PA-stimulated cardiomyocytes. Conclusion: TSG targeted to miR-129-3p/Smad3 signaling inhibited PA-induced inflammation and apoptosis in cardiomyocytes. |
| Keywords | Tetrahydroxy stilbene glucoside, miR-129-3p, Smad3, Cardiomyocytes, Inflammation, Apoptosis |
| Address and Contact Information |
1 Department of Cardiovascular Medicine, Wuhan No. 6 Hospital, Hospital Affiliated to Jianghan University, No. 168, Xianggan Road, Wuhan 430016, People’s Republic of China. 2 Department of Pharmacy, Wuhan No. 6 Hospital, Hospital Affiliated to Jianghan University, No. 168, Xianggan Road, Wuhan 430016, People’s Republic of China * Corresponding Author: kong_minfh@sina.com |
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| DOI: 10.1186/s11658-019-0142-4 Volume 24 (2019) - 24:15 | |
| Title | HONOKIOL ALLEVIATES SEPSIS-INDUCED ACUTE KIDNEY INJURY IN MICE BY TARGETING THE miR-218-5p/HEME OXYGENASE-1 SIGNALING PATHWAY |
| Authors | Tao Zhang1*† and Lei Xiang2† |
| Abstract | Background: Honokiol is a low-molecular-weight natural product and has been reported to exhibit anti-inflammatory activity. Objectives: Our study aimed to investigate the influence of honokiol on sepsis-induced acute kidney injury (AKI) in a mouse model. Material and methods: A cecal ligation and puncture (CLP) surgical operation was performed to establish a sepsis-induced acute kidney injury model in mice. Renal histomorphological analysis was performed with periodic acid-Schiff (PAS) staining. The levels of inflammatory markers in serum were measured by ELISA assay. The mRNA and protein levels were assayed by RT-qPCR and western blotting, respectively. Annexin V-FITC/PI staining was used to evaluate glomerular mesangial cell (GMC) apoptosis. Results: The results revealed that honokiol significantly increased the survival rate in mice undergoing a CLP operation. Inflammatory cytokines, such as TNF-α, IL-6 and IL-1β, were significantly inhibited in honokiol-treated septic mice compared with the CLP group. In addition, honokiol showed the ability to reverse CLP-induced AKI in septic mice. Furthermore, heme oxygenase-1 (HO-1) expression levels were significantly up-regulated and miR-218-5p was markedly down-regulated in honokiol-treated septic mice as compared to CLP-operated mice. Bioinformatics and experimental measurements showed that HO-1 was a direct target of miR-218-5p. In vitro experiments showed that both honokiol and miR-218-5p inhibitors blocked lipopolysaccharide (LPS)-induced cell growth inhibition and GMC apoptosis by increasing the expression of HO-1. Conclusions: Honokiol ameliorated AKI in septic mice and LPS-induced GMC dysfunction, and the underlying mechanism was mediated, at least partially, through the regulation of miR-218-5p/HO-1 signaling. |
| Keywords | Honokiol, Sepsis, Acute kidney injury, Heme oxygenase-1 |
| Address and Contact Information |
1 Department of of Intensive Care Unit, Tianjin Huanhu Hospital, No. 6 Jizhao Road, Tianjin 300060, People’s Republic of China 2 Department of Neurology, Tianjin Huanhu Hospital, Tianjin 300060, People’s Republic of China. * Corresponding Author: zhang_taohh@163.com † Tao Zhang and Lei Xiang contributed equally. |
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| DOI: 10.1186/s11658-019-0144-2 Volume 24 (2019) - 24:16 | |
| Title | CHONDROGENIC DIFFERENTIATION OF BONE MARROW-DERIVED MESENCHYMAL STEM CELLS FOLLOWING TRANSFECTION WITH INDIAN HEDGEHOG AND SONIC HEDGEHOG USING A ROTARY CELL CULTURE SYSTEM |
| Authors | Liyang Chen, Gejun Liu, Wenjun Li and Xing Wu* |
| Abstract | Background: Indian hedgehog (IHH) and Sonic hedgehog (SHH) are important regulators of chondrogenesis. However, activation of IHH and SHH also promotes chondrocyte hypertrophy and ossification during chondrogenesis. The aims of this study were to investigate the effect of microgravity on IHH- and SHH-induced chondrogenic differentiation and to elucidate the role of microgravity in this process. Methods: Adenovirus plasmids encoding the rabbit IHH gene and SHH genes were constructed in vitro and transfected into rabbit bone marrow-derived mesenchymal stem cells (BMSCs). A rotary cell culture system (RCCS), in which a dynamic three-dimensional culture system combines the mechanical environment with a three-dimensional culture surface, was used for cell culture and differentiation. During the induction of differentiation, expression levels of cartilage-related and cartilage hypertrophy-related genes and proteins were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting, respectively. Toluidine blue and collagen II immunohistochemical staining and annexin V-Cy3 staining were used to indicate investigate cartilage matrix synthesis and hypertrophic hypertrophy, respectively, on day 21 after induction of differentiation. Results: In this study, IHH and SHH were shown to be equipotent inducers of chondrogenesis in rabbit BMSCs, as evidenced by strong staining for proteoglycans and collagen II, and increased expression of mRNAs and proteins associated with chondrogenesis in an RCCS environment. More importantly, chondrogenic hypertrophy and aging were effectively inhibited in the RCCS environment. In addition, levels of cartilage-related markers in the IHH and SHH transfection groups were initially increased and later decreased in the traditional two-dimensional environment, while cartilage hypertrophy-related factors revealed higher mRNA expression levels during induction. Conclusions: In summary, microgravity significantly promoted chondrogenic differentiation of BMSCs induced by IHH and SHH and attenuated chondrogenic hypertrophy and aging during chondrogenesis. Furthermore, exogenous IHH and SHH had the same effect on chondrogenic differentiation of BMSCs in the RCCS environment. This study provides further evidence of chondrogenic induction of BMSCs in vitro via IHH and SHH gene delivery. |
| Keywords | Indian hedgehog, Sonic hedgehog, RCCS, Chondrogenic differentiation, Chondrocyte hypertrophy |
| Address and Contact Information | Department of Orthopedics, Shanghai Tenth People’s Hospital, School of Medicine, Tongji University, Shanghai 200072, People’s Republic of China * Corresponding author: wxing123@yeah.net |
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| DOI: 10.1186/s11658-019-0141-5 Volume 24 (2019) - 24:17 | |
| Title | TETRAMETHYLPYRAZINE EXERTS A PROTECTIVE EFFECT AGAINST INJURY FROM ACUTE MYOCARDIAL ISCHEMIA BY REGULATING THE PI3K/Akt/GSK-3β SIGNALING PATHWAY |
| Authors | Qing Yang1†, Dan Dan Huang2†, Da Guang Li1, Bo Chen1, Ling Min Zhang1, Cui Ling Yuan1 and Hong Hong Huang3* |
| Abstract | Objective: We investigated the protective effect of tetramethylpyrazine (TMP) on injury related to acute myocardial ischemia (AMI) induced by isoproterenol (ISO). Materials and methods: Rats were randomly assigned to five groups: control, ISO, ISO + propranolol (10 mg/kg), ISO + TMP (10 mg/kg) and ISO + TMP (20 mg/kg). The rats in the three ISO + groups were pretreated with propranolol or TMP, while the rats in the control and ISO groups were pretreated with an equal volume of saline. Afterwards, the rats in the four administration groups were subcutaneously injected with ISO for two consecutive days. The levels of creatine kinase (CK), lactate dehydrogenase (LDH), superoxide dismutase (SOD), malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β in the serum were measured using ELISA. The expressions of B-cell lymphoma-associated X-2 (Bax-2), B-cell lymphoma-2 (Bcl-2), phosphoinositide-3-kinase (PI3K), protein kinase B (Akt), glycogen synthase kinase 3β (GSK-3β), MDA5 and SOD1 were determined using western blotting assay. The phosphorylation of PI3K, Akt and GSK-3β were also determined using western blotting assay. The left ventricles of the rats were extracted and stained using hematoxylin and eosin (H&E). The ST segment was recorded using electrocardiograms (ECGs). Results: Administration of TMP (10, 20 mg/kg) reduced the levels of MDA and CK and the activities of SOD and LDH in the serum. Pretreatment with TMP significantly reduced the levels of pro-inflammatory cytokines, including IL-1β, IL-6 and TNF-α. Treatment with TMP also improved the histopathological alteration and decreased the ST elevation. Furthermore, TMP ameliorated the expressions of Cu, SOD1, MDA5, Bax-2, Bcl-2, p-PI3K, p-Akt and p-GSK-3β in ISO-induced rats. Conclusions: Tetramethylpyrazine protected against injury due to AMI by regulating the PI3K/Akt /GSK-3β signaling pathway. |
| Keywords | Tetramethylpyrazine, Myocardial ischemia, PI3K, Akt, GSK-3β |
| Address and Contact Information |
1 Blood Transfusion Department, First Hospital of Jilin University, Changchun, Jilin, China.
2 Preclinical School of North Sichuan Medical College, Nanchong, Sichuan, China.
3 Faculty of Chinese Medical Science, Guangxi University of Chinese Medicine, No. 13 Wuhe Road, Qingxiu District, Nanning 530222, Guangxi, China * Corresponding author: huanghongh1@foxmail.com †Qing Yang and Dan Dan Huang contributed equally to this work. |
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| DOI: 10.1186/s11658-018-0127-8 Volume 24 (2019) - 24:1 | |
| Title | MiR-106a DIRECTLY TARGETS LIMK1 TO INHIBIT PROLIFERATION AND EMT OF ORAL CARCINOMA CELLS |
| Authors | Bingxia Shi1* , Chao Ma2, Guolin Liu1 and Yanjun Guo1 |
| Abstract | Background: LIM kinase 1 (LIMK1) expression levels are closely associated with microRNA (miRNA) processing. Higher levels of LIMK1 are reported during the progression of many cancers. Our study explored the interaction between LIMK1 and miR-106a in oral squamous cell carcinoma (OSCC). Methods: Quantitative RT-PCR was performed to detect the levels of LIMK1 and miR-106a in OSCC tissues and cell lines. The rates of cell proliferation and epithelial– mesenchymal transition (EMT) were assessed to determine the biological functions of miR-106a and LIMK1 in OSCC cells. The mRNA and protein levels of LIMK1 were measured using quantitative RT-PCR and western blotting. Luciferase assays were performed to validate LIMK1 as an miR-106a target in OSCC cells. Results: We found that the level of miR-106a significantly decreased and the expression of LIMK1 significantly increased in OSCC tissues and cell lines. There was a close association between these changes. Knockdown of LIMK1 significantly inhibited the proliferation and EMT of OSCC cells. The bioinformatics analysis predicted that LIMK1 is a potential target gene of miR-106a and the luciferase reporter assay confirmed that miR-106a could directly target LIMK1. Introduction of miR-106a to OSCC cells had similar effects to LIMK1 silencing. Overexpression of LIMK1 in OSCC cells partially reversed the inhibitory effects of the miR-106a mimic. Conclusion: MiR-106a inhibited the cell proliferation and EMT of OSCC cells by directly decreasing LIMK1 expression. |
| Keywords | Oral squamous carcinoma, MicroRNA-106a, LIM kinase 1, Proliferation, Epithelial–mesenchymal transition |
| Address and Contact Information |
1 Oral and Maxillofacial Surgery, Cangzhou Central Hospital, No. 16 Xinhua West Road, Hebei 061000, People’s Republic of China 2 Department of Medical Plastic Surgery, Cangzhou Central Hospital, Hebei 061000, People’s Republic of China. * Corresponding author: shibingxiacangzhou@163.com |
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| DOI: 10.1186/s11658-019-0139-z Volume 24 (2019) - 24:19 | |
| Title | DOWNREGULATION OF CDKL1 SUPPRESSES NEUROBLASTOMA CELL PROLIFERATION, MIGRATION AND INVASION |
| Authors | Weiyi Li1, Jing Cao2, Jian Liu1, Wenli Chu1, Congqing Zhang1, Shuiling Chen1 and Zefeng Kang1* |
| Abstract | Background: Cyclin-dependent kinase-like 1 (CDKL1) is a member of the cell division control protein 2-related serine–threonine protein kinase family. It is known to occur in various malignant tumors, but its role in neuroblastoma (NB) remains unclear. Methods: We constructed a CDKL1-silenced NB cell strain (SH-SY5Y) and used real- time PCR and western blotting to confirm the silencing. Functional analyses were performed using the MTT, colony-formation, FACS, wound-healing and transwell invasion assays. Results: The expression of CDKL1 was significantly upregulated in NB tissue as compared to the adjacent normal tissue. CDKL1 knockdown significantly suppressed cell viability and colony formation ability. It also induced cell cycle G0/G1 phase arrest and apoptosis, and suppressed the migration and invasion ability of SH-SY5Y cells. CDKL1 knockdown decreased the CDK4, cyclin D1 and vimentin expression levels, and increased the caspase-3, PARP and E-cadherin expression levels in SH-SY5Y cells. Conclusions: Our findings suggest that CDKL1 plays an important role in NB cell proliferation, migration and invasion. It might serve as a potential target for NB therapy. |
| Keywords | Neuroblastoma, SH-SY5Y, CDKL1, Proliferation, Migration, Invasion |
| Address and Contact Information |
1 Eye Hospital, China Academy of Chinese Medical Sciences, No 33 Lugu Road, Shijingshan district, Beijing 100040, China 2 Yinan Branch of Qilu Hospital of Shandong University, Linyi, Shandong, China. * Corresponding author: zefeng_K2016@126.com |
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| DOI:10.1186/s11658-018-0131-z Volume 24 (2019) - 24:9 | |
| Title | miRNA-221 PROMOTES CUTANEOUS SQUAMOUS CELL CARCINOMA PROGRESSION BY TARGETING PTEN |
| Authors | Zhen-Hua Gong1†, Feng Zhou2†, Chao Shi3, Tie Xiang1, Chang-Kai Zhou1, Qian-Qian Wang1, Ya-Su Jiang1 and Sheng-Feng Gao1* |
| Abstract | Background: Cutaneous squamous cell carcinoma (CSCC) is a common type of skinmalignancy. MicroRNA-221 (miRNA-221) is a critical non-coding RNA in tumor initiationand progression. However, the molecular mechanisms of miRNA-221 in the developmentof CSCC remain unknown. This study investigated the expression of miRNA-221 in CSCCand its potential tumor biological functions. Methods: MTT assay, colony assay, PCR, and Western blot were adopted. Results: In this study, miRNA-221 expression was significantly higher in CSCC tissuesand cell lines than in normal tissues and cells (P < 0.05). Further functional experimentsindicated that miRNA-221 knockdown inhibited the proliferation and cell cycle, whileupregulation of miRNA-221 presented the opposite role. The dual reporter gene assaysindicated that PTEN is a direct target gene of miRNA-221. PTEN protein or mRNA levelswere decreased after the cells were transfected with miR-221 mimics. Conclusions: Taken together, the obtained results indicated that miR-221 plays anoncogenic function in CSCC by targeting PTEN and further suggest that miR-221 maybe a potential target for CSCC diagnosis and treatment. |
| Keywords | Cutaneous squamous cell carcinoma, miRNA-221, PTEN, Proliferation, Treatment |
| Address and Contact Information | 1 Department of Burn and Plastic Surgery, The First People’s Hospital of Nantong, Nantong 226001, China 2 Department of Clinical Laboratory, The First People’s Hospital of Nantong, Nantong 226001, China. 3 Department of Pathology, The First People’s Hospital of Nantong, Nantong 226001, China. * Corresponding author: 574482029@qq.com † Zhen-Hua Gong and Feng Zhou contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0145-1 Volume 24 (2019) - 24:20 | |
| Title | INHIBITION OF miR-9-5p SUPPRESSES PROSTATE CANCER PROGRESS BY TARGETING StarD13 |
| Authors | Lin Chen*, Weifeng Hu, Guohao Li, Yonglian Guo, Zhihua Wan and Jiajun Yu |
| Abstract | Background: This study aims to investigate the effects of inhibiting microRNA-9-5p (miR-9-5p) on the expression of StAR-related lipid transfer domain containing 13 (StarD13) and the progress of prostate cancer. Methods: The mRNA expression levels of miR-9-5p and StarD13 were determined in several prostate cancer cell lines. We chose DU145 and PC-3 cells for further research. The CCK8 assay was used to measure the cell viability. The cell invasion and wound-healing assays were respectively applied to evaluate invasion and migration. The expression of E-cadherin (E-cad), N-cadherin (N-cad) and vimentin were measured via western blot. DU145 and PC-3 cells overexpressing StarD13 were generated to investigate the variation in proliferation, invasion and migration. A luciferase reporter assay was used to identify the target of miR-9-5p. Results: Our results show that miR-9-5p was highly expressed and StarD13 was suppressed in prostate cancer cells. MiR-9-5p inhibition repressed the cells’ viability, invasion and migration. It also increased the expression of E-cad and decreased that of N-cad and vimentin. StarD13 overexpression gave the same results as silencing of miR-9-5p: suppression of cell proliferation, invasion and migration. The bioinformatics analysis predicted StarD13 as a target gene of miR-9-5p. Quantitative RT-PCR, western blot analysis and the dual-luciferase reporter assay were employed to confirm the prediction. Conclusion: Our results show that miR-9-5p plays a powerful role in the growth, invasion, migration and epithelial–mesenchymal transition (EMT) of prostate cancer cells by regulating StarD13. A therapeutic agent inhibiting miR-9-5p could act as a tumor suppressor for prostate cancer. |
| Keywords | microRNA-9-5p, Prostate cancer, StarD13, Migration, Invasion |
| Address and Contact Information | Department of Urology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, No. 26 Shengli Street, Jiang’an District, Wuhan 430014, China * Corresponding author: chenlin8279@yeah.net |
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| DOI: 10.1186/s11658-018-0134-9 Volume 24 (2019) - 24:10 | |
| Title | SUPPRESSING microRNA-29c PROMOTES BILIARY ATRESIA-RELATED FIBROSIS BY TARGETING DNMT3A AND DNMT3B |
| Authors | Jian-yao Wang1, Hao Cheng2, Hong-yan Zhang2, Yong-qin Ye1, Qi Feng1, Zi-min Chen1, Yue-lan Zheng1, Zhou-guang Wu1, Bin Wang1* and Jun Yao3* |
| Abstract | This study was designed to investigate the potential role of microRNA-29c (miR-29c) in biliary atresia-related fibrosis. The expression of miR-29c was determined in 15 pairs of peripheral blood samples from infants with biliary atresia (BA) and infants with non-BA neonatal cholestasis using quantitative real-time PCR. EMT was established by induction with TGF-β1 in HIBEpiC cells. MiR-29c was inhibited by lipofectamine transfection. The expressions of proteins related to epithelial–mesenchymal transition (EMT), i.e., E-cadherin, N-cadherin and vimentin, were determined using quantitative real-time PCR and western blotting. Direct interaction between miR-29c and DNMT3A and DNMT3B was identified using a luciferase reporter assay. The expressions of DNMT3A and DNMT3B were suppressed by treatment with SGI-1027. Patients with BA showed significantly lower miR-29c levels in peripheral blood samples than the control subjects. In vitro, TGF-β1-induced EMT significantly decreased the expression of miR-29c. Downregulation of miR-29c had a promotional effect on BA-related fibrosis in HIBEpiC cells, as confirmed by the decrease in E-cadherin and increase in N-cadherin and vimentin levels. MiR-29c was found to target the 3’UTR of DNMT3A and DNMT3B and inhibit their expression. Suppression of DNMT3A and DNMT3B reversed the effects of miR-29c downregulation on BA-related fibrosis in HIBEpiC cells. These data suggest that BA-related fibrosis is closely associated with the occurrence of EMT in HIBEpiC cells. MiR-29c might be a candidate for alleviating BA-related fibrosis by targeting DNMT3A and DNMT3B. |
| Keywords | Biliary atresia, Epithelial–mesenchymal transition, MiR-29c, Fibrosis, DNMT3A, DNMT3B |
| Address and Contact Information |
1 Department of General Surgery, Shenzhen Children’s Hospital, Shenzhen 518026, Guangdong Province, China 2 Graduate School of China Medical University, Shenzhen 110122, Liaoning Province, China. 3 Department of Gastroenterology, Jinan University of Medical Sciences, Shenzhen Municipal People’s Hospital, Shenzhen 518020, Guangdong Province, China * Corresponding author: szwb1967@126.com; yj_1108@126.com |
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| DOI: 10.1186/s11658-019-0147-z Volume 24 (2019) - 24:21 | |
| Title | MELATONIN RECEPTOR DEPLETION SUPPRESSED hCG-INDUCED TESTOSTERONE EXPRESSION IN MOUSE LEYDIG CELLS |
| Authors | Yuan Gao1*†, Xiaochun Wu2†, Shuqin Zhao1, Yujun Zhang1, Hailong Ma1, Zhen Yang1, Wanghao Yang1, Chen Zhao1, Li Wang1 and Quanwei Zhang1,2 |
| Abstract | Melatonin receptors MT1 and MT2 (genes officially named MTNR1A and MTNR1B, respectively) play crucial roles in melatonin-mediated regulation of circadian rhythms, the immune system, and control of reproduction in seasonally breeding animals. In this study, immunolocalization assay showed that MT1 and MT2 are highly expressed in Leydig cell membrane. To understand the biological function of melatonin receptors in hCG-induced testosterone synthesis, we generated melatonin receptor knockdown cells using specific siRNA and performed testosterone detection after hCG treatment. We found that knockdown of melatonin receptors, especially MTNR1A, led to an obvious decrease (> 60%) of testosterone level. Our further study revealed that knockdown of melatonin receptors repressed expression, at both the mRNA level and the protein level, of key steroidogenic genes, such as p450scc, p450c17 and StAR, which are essential for testosterone synthesis. hCG triggered endoplasmic reticulum (ER) stress to regulate steroidogenic genes’ expression and apoptosis. To further investigate the potential roles of melatonin receptors in hCG-induced regulation of ER stress and apoptosis, we examined expression of some crucial ER stress markers, including Grp78, Chop, ATF4, Xbp1, and IRE1. We found that inhibition of melatonin receptors increased hCG-induced expression of Grp78, Chop and ATF4, but not Xbp1 and IRE1, suggesting that hCG may modulate IRE1 signaling pathways in a melatonin receptor-dependent manner. In addition, our further data showed that knockdown of MTNR1A and MTNR1B promoted hCG-induced expression of apoptosis markers, including p53, caspase-3 and Bcl-2. These results suggested that the melatonin receptors MTNR1A and MTNR1B are essential to repress hCG-induced ER stress and cell apoptosis. Our studies demonstrated that the mammalian melatonin receptors MT1 and MT2 are involved in testosterone synthesis via mediating multiple cell pathways. |
| Keywords | Melatonin receptor, Testosterone, ER stress, Apoptosis |
| Address and Contact Information |
1 College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, Gansu, China2 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu, China. * Corresponding author: yyshpy@gmail.com †Yuan Gao and Xiaochun Wu contributed equally to this work. Yuan Gao and Xiaochun Wu are considered co-first authors. |
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| DOI: 10.1186/s11658-019-0136-2 Volume 24 (2019) - 24:11 | |
| Title | miR-29b-3p REGULATED OSTEOBLAST DIFFERENTIATION VIA REGULATING IGF-1 SECRETION OF MECHANICALLY STIMULATED OSTEOCYTES |
| Authors | Qiangcheng Zeng1†, Yang Wang2,3†, Jie Gao1,4, Zhixiong Yan2, Zhenghua Li1, Xianqiong Zou2, Yanan Li2, Jiahui Wang2 and Yong Guo1,2* |
| Abstract | Background: Mechanical loading is an essential factor for bone formation. A previous study indicated that mechanical tensile strain of 2500 microstrain (με) at 0.5 Hz for 8 h promoted osteogenesis and corresponding mechanoresponsive microRNAs (miRs) were identified in osteoblasts. However, in osteocytes, it has not been identified which miRs respond to the mechanical strain, and it is not fully understood how the mechanoresponsive miRs regulate osteoblast differentiation. Methods: Mouse MLO-Y4 osteocytes were applied to the same mechanical tensile strain in vitro. Using molecular and biochemical methods, IGF-1 (insulin-like growth factor-1), PGE2 (prostaglandin E2), OPG (osteoprotegerin) and NOS (nitric oxide synthase) activities of the cells were assayed. MiR microarray and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assays were applied to select and validate differentially expressed miRs, and the target genes of these miRs were then predicted. MC3T3-E1 osteoblasts were stimulated by the mechanical tensile strain, and the miR-29b-3p expression was detected with miR microarray and RT-qPCR. Additionally, the effect of miR-29b-3p on IFG-1 secretion of osteocytes and the influence of conditioned medium of osteocytes transfected with miR-29b-3p on osteoblast differentiation were investigated. Results: The mechanical strain increased secretions of IGF-1 and PGE2, elevated OPG expression and NOS activities, and resulted in altered expression of the ten miRs, and possible target genes for these differentially expressed miRs were revealed through bioinformatics. Among the ten miRs, miR-29b-3p were down-regulated, and miR-29b-3p overexpression decreased the IGF-1 secretion of osteocytes. The mechanical strain did not change expression of osteoblasts’ miR-29b-3p. In addition, the conditioned medium of mechanically strained osteocytes promoted osteoblast differentiation, and the conditioned medium of osteocytes transfected with miR-29b-3p mimic inhibited osteoblast differentiation. Conclusions: In osteocytes (but not osteoblasts), miR-29b-3p was responsive to the mechanical tensile strain and regulated osteoblast differentiation via regulating IGF-1 secretion of mechanically strained osteocytes. |
| Keywords | Mechanical tensile strain, Osteocyte, Osteoblast differentiation, miRNA microarray |
| Address and Contact Information |
1 key laboratory of Functional Bioresource Utilization in University of Shandong, Shandong Key Laboratory of Biophysics, Dezhou University, Dezhou 253023, China. 2 Department of Biomedical Engineering, College of Biotechnology, Guilin Medical University, No. 1 Zhiyuan Road, Lingui District, Guilin City 541100, Guangxi, China China. 3 Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, Bioengineering College of Chongqing University, Chongqing 400044, China. 4 Medical Department, Secondary Renmin Hospital of Dezhou, Dezhou 253023, Shangdong, China. * Corresponding author: guoyong74@163.com †Qiangcheng Zeng and Yang Wang contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0143-3 Volume 24 (2019) - 24:22 | |
| Title | MiR-214 SENSITIZES HUMAN COLON CANCER CELLS TO 5-FU BY TARGETING Hsp27 |
| Authors | Yong Yang1,2, Yan Bao1,2, Guo-Kai Yang1,2, Jia Wan1,2, Ling-Juan Du1,2 and Zhen-Huan Ma1,2* |
| Abstract | Overcoming chemorestistance to 5-fluorouracil (5-FU) could offer a new treatment option for highly malignant colon cancer. In our study, differential microRNA expression profiling revealed that miR-214 is downregulated in 5-FU-resistant colon cancer cells compared to normal cells. In vitro, miR-214 could sensitize non-resistant colon cancer cells and 5-FU-resistant colon cancer cellsto 5-FU. Functionally, miR-214 inhibited cell clone formation and cell growth and enhanced 5-FU-inducing cell apoptosis and caspase-3 levels. MiR-214 targeted heat shock protein 27 (Hsp27), as confirmed via dual luciferase reporter assays and western blots. Hsp27 also sensitized HT-29 and LoVo to 5-FU by enhancing cell apoptosis. Overexpression of Hsp27 could block miR-214 with an effect on the sensitivity of colon cancer cells to 5-FU. In conclusion, miR- 214 sensitizes colon cancer cells to 5-FU by targeting Hsp27, indicating a significant role for this miRNA in colon cancer chemotherapy. |
| Keywords | miR-214, Hsp27, 3′-UTR, 5-FU, Colon cancer |
| Address and Contact Information | 1 The Third Department of General Surgery, The Second People’s Hospital of Yunnan Province, Kunming, China 2 Department of Vascular Surgery, The Fourth Affiliated Hospital of Kunming MedicalUniversity, 176 Youth Road, Kunming, Yunnan Province 650021, People’s Republic of China * Corresponding author: yyyunnan@163.com |
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| DOI: 10.1186/s11658-019-0138-0 Volume 24 (2019) - 24:12 | |
| Title | THE Pro12Ala POLYMORPHISM IN THE PPARγ2 GENE IS NOT ASSOCIATED WITH AN INCREASED RISK OF NAFLD IN IRANIAN PATIENTS WITH TYPE 2 DIABETES MELLITUS |
| Authors | Leila Saremi1, Shirin Lotfipanah2, Masumeh Mohammadi1, Hassan Hosseinzadeh3, Mina Fathi-Kazerooni4, Behrooz Johari5 and Zohreh Saltanatpour6* |
| Abstract | Background: The peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors that belong to the nuclear hormone receptor superfamily. Several studies have demonstrated a significant association between Pro12Ala polymorphism of the PPARγ2 gene and metabolic disorders. Therefore, this study aimed to evaluate the association of Pro12Ala polymorphism with increased risk of NAFLD in Iranian patients with type 2 diabetes mellitus. Methods: This cross-sectional study was performed on 145 healthy control subjects and 145 NAFLD patients with a history of type 2 diabetes. Pro12Ala polymorphism genotyping was performed using PCR–restriction fragment length polymorphism (RFLP) technique with the Bs1I restriction enzyme. Results: Our results demonstrated that CC and GG genotypes of Pro12Ala were found in the participants, but there was no statistically significant difference between NAFLD patients and healthy controls (P = 0.64 and χ2 = 0.21). Conclusion: This study suggests that Pro12Ala polymorphism of the PPARγ2 gene cannot be considered as a risk factor for NAFLD in the Iranian population. |
| Keywords | Pro12Ala polymorphism, PPARγ2 gene, NAFLD, Type 2 diabetes mellitus |
| Address and Contact Information |
1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. 2 Farhangian University, Shahid Mofatteh Teacher Education Paradise, Tehran, Iran. 3 Department of Biology, Faculty of Science, Yazd University, Yazd, Iran. 4 Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran. 5 Department of Medical Biotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran. 6 Medical Genetic Center, Endocrinology and Metabolism Research Institute (EMRI), Tehran University of Medical Sciences (TUMS), Tehran, Iran. * Corresponding author: z-saltanatpour@razi.tums.ac.ir; zohre_saltanatpour@yahoo.com |
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| DOI: 10.1186/s11658-018-0132-y Volume 24 (2019) - 24:2 | |
| Title | miR-573 REGULATES CELL PROLIFERATION AND APOPTOSIS BY TARGETING BAX IN NUCLEUS PULPOSUS CELLS |
| Authors | Rui Wang1, Boping Wen2 and Dong Sun1* |
| Abstract | Background: MicroRNA (miRNA) plays a vital role in the pathogenesis of intervertebral disc degeneration (IDD). The expression and potential mechanism of miR-573 in human nucleus pulposus (NP) remains to be elucidated. In this study, we aimed to investigate the role of miR-573 in IDD. Methods: Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis was applied to examine the expression of miR-573 and Bax in idiopathic scoliosis tissues and IDD tissues. Human NP cells were employed for analysis. Moreover, the proliferation and apoptosis of NP cells were detected using MTT and flow cytometry assay respectively. The expression levels of Bcl-2, cleaved caspase-3, cleaved caspase-9, caspase-3 and caspase-9 in degenerative NP cells were measured by Western blotting assay. Furthermore, a luciferase reporter assay was used to verify the relationship between miR-573 and Bax. Results: The results revealed that the mRNA expression level of miR-573 was down- regulated whereas Bax was up-regulated notably in degenerative NP cells. In addition, overexpression of miR-573 increased cell viability remarkably, coupled with inhibition of cell apoptosis. The expression level of Bcl-2 was increased while cleaved caspase-3 and cleaved caspase-9 expression levels were decreased in miR-573 overexpression NP cells. Additionally, the bioinformatics analysis underscored that Bax was a direct target gene of miR-573. Conclusion: These results suggest that overexpression of miR-573 inhibited NP cell apoptosis by down-regulating Bax, which proved to be a novel effective strategy for IDD therapies. |
| Keywords | miR-573, Bax, Nucleus pulposus cells, Intervertebral disc degeneration |
| Address and Contact Information |
1 Department of Massage and Physiotherapy, Guang Xing Hospital, Zhejiang University of Traditional Chinese Medicine, No. 453, Tiyuchang Road, Xihu District, Hangzhou, Zhejiang 310007, People’s Republic of China 2 Department of Rehabilitation, Western Theater General Hospital, Chengdu, Sichuan 610011, People’s Republic of China. * Corresponding author: sundonghz@126.com |
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| DOI: 10.1186/s11658-019-0135-3 Volume 24 (2019) - 24:18 | |
| Title | HYPOXIA PROTECTS AGAINST THE CELL DEATH TRIGGERED BY OXOVANADIUM–GALACTOMANNAN COMPLEXES IN HepG2 CELLS |
| Authors | Monique Meyenberg Cunha-de Padua1,2,3, Guilhermina Rodrigues Noleto1*, Carmen Lucia de Oliveira Petkowicz1, Silvia Maria Suter Correia Cadena1, Frédéric Bost3, Jacques Pouysségur2,4 and Nathalie M. Mazure2,3* |
| Abstract | Background: Polysaccharides from various sources have been used in traditional medicine for centuries. The beneficial pharmacological effects of plant-derived polysaccharides include anti-tumor activity. Methods: Here, we evaluated the anti-cancer effect of the MSAGM:VO complex under hypoxic conditions (1% oxygen). MSAGM:VO is a complex of the hydrolysate of galactomannan (MSAGM) from Schizolobium amazonicum with oxovanadium (IV/V). The hepatocellular carcinoma (HCC) cell line HepG2 was selected as HCC are one of the most hypoxic solid tumors. Results: Our results showed that the strong apoptotic activity of MSAGM:VO observed in HepG2 cells under normoxic conditions was completely lost under hypoxic conditions. We found a dynamic balance between the pro- and anti-apoptotic members of the Bcl-2 protein family. The expressions of anti-apoptotic Mcl-1 and Bcl-XL increased in hypoxia, whereas the expression of pro-apoptotic Bax decreased. MSAGM:VO strongly induced autophagy, which was previously characterized as a pro-survival mechanism in hypoxia. These results demonstrate total elimination of the anti-cancer activity of MSAGM:VO with activation of autophagy under conditions of hypoxia. Conclusion: Although this study is a proof-of-concept of the impact of hypoxia on the potential of polysaccharides, further study is encouraged. The anti-tumor activity of polysaccharides could be achieved in normoxia or through raising the activity of the immune system. In addition, combination strategies for therapy with anti-autophagic drugs could be proposed. |
| Keywords | Hepatocellular carcinoma, Hypoxia, MSAGM:VO, Polysaccharides |
| Address and Contact Information |
1 Department of Biochemistry and Molecular Biology, Federal University of Parana, Curitiba, Brazil 2 Institute for Research on Cancer and Aging of Nice, CNRS-UMR 7284-Inserm U1081, University of Nice Sophia-Antipolis, Centre Antoine Lacassagne, 33 Ave. de Valombrose, 06189 Nice, France 3 Present Address: INSERM U1065, C3M, 151 Route de St Antoine de Ginestière, BP2 3194, 06204 Nice Cedex 03, France. 4 Medical Biology Department, Centre Scientifique de Monaco (CSM), Monaco, Monaco. * Corresponding author: guinoleto@yahoo.com.br; mazure@unice.fr |
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| DOI: 10.1186/s11658-019-0146-0 Volume 24 (2019) - 24:23 | |
| Title | MiR-125a-5p PROMOTES OSTEOCLASTOGENESIS BY TARGETING TNFRSF1B |
| Authors | Liang Sun, Jun Xiang Lian and Shu Meng* |
| Abstract | Aim: To investigate the dysregulation of microRNAs (miRNAs) during the differentiation of osteoclasts and the precise roles of miR-125a-5p in the differentiation of osteoclasts. Methods: The cell model of RAW 264.7 osteoclast precursor cell differentiation induced by RANKL plus M-CSF stimulation was established. During the early stage of osteoclast differentiation, miRNA expression profiles were detected using the biochip technique and analyzed by cluster analysis. TargetScan, miRTarBase and miRDB database analysis was applied to find the key target genes of miR-125a-5p. A dual luciferase experiment was conducted to identify the direct target of miR-125a-5p. MiR-125a-5p mimic transfection and anti-miR-125-5p treatment were conducted to verify the role of miR-125q-5p in osteoclast differentiation. The levels of triiodothyronine receptor auxiliary protein (TRAP), matrix metallopeptidase 2 (MMP-2), MMP-9 and cathepsin K were analyzed by qRT-PCR and western blot assay. The expression levels of MMP-2 and MMP-9 were determined using western blotting and immunofluorescence assay. The migration and invasion of RAW 264.7 cells were assessed by wound healing and Transwell invasion assays. The proliferation of RAW 264.7 osteoclast precursor cells was detected using MTT assay. Results: There were 44 microRNAs differently expressed during the differentiation of RAW 264.7 osteoclast precursor cells into osteoclasts, 35 of which were up-regulated and 9 were down-regulated. By luciferase reporter assay, it was confirmed that the TNF receptor superfamily member 1B gene (TNFRSF1B) was the target gene of miR-125a-5p. Up-regulation of miR-125a-5p inhibited TNFRSF1B protein expression and promoted osteoclast differentiation whereas down-regulation of miR-125a-5p caused completely opposite results. Conclusions: In conclusion, overexpression of miR-125a-5p suppresses the expression of TNFRSF1B and promotes osteoclast differentiation. These results reveal the crucial role of miR-125a-5p in the differentiation of osteoclasts. |
| Keywords | miR-125a-5p, Osteoclast, Osteoporosis, TNFRSF1B |
| Address and Contact Information | State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, No. 14, Section 3 of RenMinNanlu, Chengdu 610041, Sichuan, China * Corresponding author: mengshumens@foxmail.com |
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| DOI: 10.1186/s11658-019-0149-x Volume 24 (2019) - 24:24 | |
| Title | HYPOXIA-ASSOCIATED circDENND2A PROMOTES GLIOMA AGGRESSIVENESS BY SPONGING miR-625-5p |
| Authors | Hui Su1,2, Defei Zou1,2, Yikun Sun3 and Yiwu Dai1,2* |
| Abstract | Background: As a newfound type of non-coding RNA, circular RNAs (circRNAs) are involved in various physiological and pathological processes via regulation of gene expression. Increasing evidence shows that aberrantly expressed circRNAs play a crucial role in the initiation and progression of many tumors. However, the functions of different circRNAs in gliomas remain elusive. Methods: The levels of circRNAs, miRNAs, and mRNAs were quantified by qPCR. The interaction between circDENND2A and miR-625-5p was determined by luciferase reporter and pull-down assays. The migratory and invasive capabilities of glioma cells were examined by wound healing and Transwell assays. Immunohistochemistry was performed to analyze the HIF1α level in glioma tissues. Results: We predicted circDENND2A (has_circ_0002142) to be a hypoxia-responsive circRNA in glioma via a bioinformatic analysis. We found that hypoxia induced the expression of circDENND2A, which promoted migration and invasion of glioma cells. To understand the behaviors of circDENND2A in glioma, we studied the putative miRNAs targeted by circDENND2A and identified circDENND2A as an efficient sponge of miR-625-5p in glioma cells. Phenotype experiments verified that circDENND2A was required for the hypoxia-induced migration and invasion of glioma cells and that this occurred by sponging of miR-625-5p. Notably, glioma tissues overexpressing HIF1α exhibited a high expression of circDENND2A as well as a low expression of miR-625-5p. circDENND2A was negatively correlated with miR-625-5p. Conclusion: circDENND2A is required for the hypoxia-induced malignancy of glioma cells and functions by sponging miR-625-5p. |
| Keywords | circDENND2A, miR-625-5p, Glioma, Hypoxia, Migration, Invasion |
| Address and Contact Information |
1 Department of Neurosurgery, Chinese PLA General Hospital, Medical School of Chinese PLA, Beijing, People’s Republic of China 2 Department of Neurosurgery, Bayi Brain Hospital, PLA Army General Hospital, Beijing, People’s Republic of China 3 Department of Neurosurgery, The 306th Hospital of PLA, Beijing, People’s Republic of China. * Correspondng author: dddyyywww@163.com |
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| DOI: 10.1186/s11658-019-0150-4 Volume 24 (2019) - 24:25 | |
| Title | THE ROLE OF miR-431-5p IN REGULATING PULMONARY SURFACTANT EXPRESSION IN VITRO |
| Authors | Shujun Li1†, Zhongyi Sun2†, Tao Chen3, Jingjing Pan2, Yanqing Shen4, Xiaoqing Chen2, Xiaoyu Zhou4, Rui Cheng4 and Yang Yang4* |
| Abstract | Background: Pulmonary surfactant is the complex mixture of lipid and protein that covers the alveolar surface. Pulmonary surfactant deficiency is one of the main causes of neonatal respiratory distress. Recent studies showed that miRNA plays an important role in lung development, but research into miR-431 regulation of pulmonary surfactant are sparse. In this study, we explored the regulatory role of miR-431-5p in the expression of pulmonary surfactant and identified its potential target gene, Smad4. Methods: The bioinformatics tool TargetScan was used to predict the targets of miR-431. The expression of miR-431-5p was achieved via transfection of miR-431-5p mimics, an miR-431-5p inhibitor and corresponding negative control. The level of miR-431-5p was determined using quantitative real-time PCR. The CCK8 assay was conducted to confirm cell growth 12 h after transfection with miR-431-5p mimics, inhibitor or NC. Smad4 and surfactant-associated proteins in A549 were analyzed using western blot and quantitative real-time PCR. Results: Smad4 was validated as a target of miR-431 in A549 cells. Overexpression of miR-431 accelerated A549 proliferation and inhibited A549 apoptosis. The mRNA and protein levels for the surfactant proteins (SP-A, SP-B, SP-C and SP-D) were found to be differentially expressed in A549 cells over- or under-expressing miR-431-5p. Conclusion: Our results show that miR-431-5p is critical for pulmonary surfactant expression and that its regulation is closely related to the TGF-β/Smad4 pathway. These results will help us to study the pathophysiological mechanism of lung developmental diseases. |
| Keywords | miR-431-5p, Pulmonary surfactant, TGF-β/Smads pathway, Lung development |
| Address and Contact Information |
1 Department of Pediatrics, Children’s Hospital of Anhui Medical University, Hefei, China. 2 Department of Pediatrics, The First Affliated Hospital of Nanjing Medical University, Nanjing, China. 3 Department of Cardiothoracic Surgery, The First Affliated Hospital of Anhui Medical University, Hefei, China. 4 Department of Neonates, Children’s Hospital of Nanjing Medical University, No 72, Guangzhou Road, Nanjing 210008, China * Corresponding author: yy860507@126.com; 15952071803@163.com †Shujun Li and Zhongyi Sun contributed equally to this work. |
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| DOI: 10.1186/s11658-018-0133-x Volume 24 (2019) - 24:3 | |
| Title | LIMBAL NICHE CELLS CAN REDUCE THE ANGIOGENIC POTENTIAL OF CULTIVATED ORAL MUCOSAL EPITHELIAL CELLS |
| Authors | Chao-Ye Duan, Hua-Tao Xie, Xin-Yue Zhao, Wen-Han Xu and Ming-Chang Zhang* |
| Abstract | Background: Autologous cultivated oral mucosal epithelial transplantation (COMET) is an important treatment for limbal stem cell deficiency. However, peripheral corneal neovascularization after surgery hinders its application. This study aims to employ a culture system using allogenic limbal niche cells (LNCs) instead of mouse-derived 3T3 cells as a feeder layer that could relieve postoperative neovascularization. Methods: Rat oral mucosal epithelial cells (OMECs) were co-cultured with rat LNCs or 3T3 cells. Cultivated oral mucosal epithelial cells (COMECs) of different culture systems were identified by hematoxylin and eosin staining and immunocytochemistry. The expression levels of the angiogenesis-related factors were analyzed by RT-qPCR and western blotting/ELISA. Angiogenic potential was reconfirmed by cell viability and tube formation assays with human umbilical vein endothelial cells (HUVECs). Results: COMECs were obtained from both culture systems successfully. Immunocytochemistry showed approximately equal percentages of positive staining cells for p63α (p = 0.9177), ABCG2 (p = 0.526), Ki67 (p = 0.0987), and CK3 (p = 0.4000) in COMECs of different groups. RT-qPCR and western blotting/ELISA showed that COMECs of the LNC group expressed a significantly lower amount of basic fibroblast growth factor (bFGF) (p = 0.0038 for RT-qPCR, p = 0.0026 for western blotting) but more pigment epithelium-derived factor (PEDF) (p = 0.0172 for RT-qPCR, p = 0.0253 for western blotting) and soluble fms-like tyrosine kinase-1 (sFlt-1) (p < 0.0001 for RT-qPCR, p = 0.0064 for ELISA) than the COMECs of the 3T3 group. Furthermore, compared with COMECs of the 3T3 group, COMECs of the LNC group could reduce the viability (p = 0.0002) and tube formation (p = 0.0002) of HUVECs. Conclusions: LNCs could substitute 3T3 cells for expanding OMECs in vitro, and the COMECs obtained in this system are less likely to induce postsurgical neovascularization, which provides an alternative option for an ex vivo culture system and promotes the application of COMET. |
| Keywords | Limbal niche cells, 3T3 cells, Limbal stem cell deficiency, Oral mucosal epithelial transplantation, Corneal neovascularization |
| Address and Contact Information | Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China * Corresponding author: mingchangzhang@hotmail.com |
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| DOI: 10.1186/s11658-019-0148-y Volume 24 (2019) - 24:26 | |
| Title | THE lncRNA MIR4435-2HG IS UPREGULATED IN HEPATOCELLULAR CARCINOMA AND PROMOTES CANCER CELL PROLIFERATION BY UPREGULATING miRNA-487a |
| Authors | Qinglei Kong1†, Caiqian Liang1†, Yi Jin2*†, Yuhang Pan2, Dayue Tong3, Qingcong Kong4* and Jing Zhou2* |
| Abstract | Background: Given the high mortality rate and unclear pathogenesis for liver cancer, investigation of its molecular mechanisms is essential. We focused on the long non-coding RNA (lncRNA) MIR4435-2HG, which was recently reported to be oncogenic in lung cancer and the microRNA miRNA-487a, which has been reported to be oncogenic in hepatocellular carcinoma (HCC). Our aim was to determine if the former has a role in HCC, and to further validate the role of the latter. Methods: Samples from 64 patients with HCC were taken at The Third Affiliated Hospital of Sun Yat-Sen University. Cell transfection and PCR were applied. Results: We found that MIR4435-2HG and miRNA-487a were upregulated in tumor tissues compared to adjacent healthy tissues from HCC patients. The expression of MIR4435-2HG was significantly affected by tumor size but not by tumor metastasis. Correlation analysis showed that MIR4435-2HG and miRNA-487a were positively correlated in both the tumor tissues and adjacent healthy tissues from HCC patients. Overexpression of MIR4435-2HG led to upregulation of miRNA-487a in the cells of HCC cell lines, while overexpression of miRNA-487a did not significantly affect MIR4435-2HG. Overexpression of MIR4435-2HG and miRNA-487a promoted the proliferation of cells of HCC cell lines, and miRNA-487a knockdown partially attenuated the enhancing effects of MIR4435-2HG overexpression on cancer cell proliferation. Conclusion: MIR4435-2HG is upregulated in HCC and promotes cancer cell proliferation possibly by upregulating miRNA-487a. |
| Keywords | Hepatocellular carcinoma, lncRNA MIR4435-2HG, miRNA-487a, Proliferation |
| Address and Contact Information |
1 Department of Emergency Medicine, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou City, Guangdong Province 510630, People’s Republic of China. 2 Department of Pathology, Guangdong Provincial Key Laboratory of Liver Disease Research, The Third Affiliated Hospital, Sun Yat-Sen University, No. 600 Tianhe Road, Guangzhou City, Guangdong Province 510630, People’s Republic of China 3 Department of Forensic Medicine, ZhongShan Medical School, Sun Yat-Sen University, Guangzhou City, Guangdong Province 510080, People’s Republic of China. 4 Department of Radiology, The Third Affiliated Hospital, Sun Yat-Sen University, No. 600 Tianhe Road, Guangzhou City, Guangdong Province 510630, People’s Republic of China * Corresponding author: zk56932@163.com; zk56932@163.com; zk56932@163.com †Qinglei Kong, Caiqian Liang and Yi Jin contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0151-3 Volume 24 (2019) - 24:27 | |
| Title | UPREGULATION OF miR-29b-3p PROTECTS CARDIOMYOCYTES FROM HYPOXIA-INDUCED APOPTOSIS BY TARGETING TRAF5 |
| Authors | Yuhua Cai1 and Yunpeng Li2* |
| Abstract | Background: MicroRNAs (miRNAs) are pivotal regulators in regulating hypoxia-induced cardiomyocyte injury. This study was designed to evaluate the effects of miR-29b-3p on hypoxic cardiomyocytes. Methods: Human AC16 cells were cultured under normoxic or hypoxic conditions. Hypoxic injury was confirmed based on alterations in cell viability using CCK-8 assay and apoptosis using flow cytometry and Hoechst staining. Bioinformatics analyses and the dual-luciferase reporter assay were performed to predict and validate the target gene of miR-29b-3p. Results: We found that hypoxia suppressed cell viability and promoted apoptosis. TNF receptor-associated factor 5 (TRAF5) was a potential target gene of miR-29b-3p. Our in vitro experiments revealed that miR-29b-3p overexpression or TRAF6 knockdown significantly protected cardiomyocytes against hypoxia-induced injury. Moreover, knockdown of TRAF5 knockdown potentiated the protective effects of miR-29b-3p against hypoxia-induced cell injury. Conclusion: These findings suggest that upregulation of miR-29b-3p could protect cardiomyocytes against hypoxia-induced injury through downregulation of TRAF5. Targeting TRAF5 with miR-29b-3p might be a potential therapeutic method for AMI. |
| Keywords | Cardiomyocyte, Hypoxia, miR-29b-3p, TRAF5 |
| Address and Contact Information |
1 Department of Cardiovasology, Jingzhou First Municipal Hospital, Jingzhou, Hubei Province, China. 2 Department of Cardiovasology, Dongfeng Hospital, Hubei University of Medicine, No. 16 Daling Road, Shiyan 442008, Hubei Province, China * Corresponding author: yunP_litea@126.com |
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| DOI: 10.1186/s11658-018-0130-0 Volume 24 (2019) - 24:8 | |
| Title | MiR-613 INHIBITS PROLIFERATION AND INVASION AND INDUCES APOPTOSIS OF RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLASTS BY DIRECT DOWN-REGULATION OF DKK1 |
| Authors | Liang Liu1* , Yanhua Zuo1, Yan Xu2, Zongfang Zhang1, Ying Li1 and Jie Pang1 |
| Abstract | Background: This study aimed to investigate the effects of miR-613 on the proliferation, invasion and apoptosis of rheumatoid arthritis synovial fibroblasts (RASFs). Methods: Synovial tissue samples were collected from 20 rheumatoid arthritis (RA) patients and 10 patients with joint trauma undergoing joint replacement surgery. The RASFs were isolated and cultured. MiR-613 and DKK1 expression in both synovial tissues and cells was detected using quantitative real-time PCR (qRT-PCR). Dual luciferase reporter gene assay was employed to evaluate the effect of miR-613 on the luciferase activity of DKK1. Then RASFs were transfected with miR-613 mimics, si-DKK1 and pcDNA-DKK1. Changes in cellular proliferation, invasion and apoptosis were detected through BrdU assay, Transwell invasion assay and flow cytometry analysis, respectively. Results: MiR-613 was significantly down-regulated in RA tissues and RASFs compared to normal tissues and cells, whereas DKK1 was up-regulated in RA tissues and RASFs. Dual luciferase reporter gene assay showed that miR-613 could specifically bind to the 3′UTR of DKK1 and significantly inhibit the luciferase activity. Moreover, miR-613 significantly reduced the expression of DKK1. Overexpression of miR-613 or knockdown of DKK1 suppressed proliferation and invasion of RASFs, and induced RASF apoptosis. The reverse results were observed when DKK1 was up-regulated in miR-613-overexpressing RASFs. Conclusions: MiR-613 can inhibit proliferation and invasion and induce apoptosis of RASFs by directly targeting DKK1 expression. |
| Keywords | Rheumatoid arthritis, MicroRNA-613, Rheumatoid arthritis synovial fibroblasts, DKK1, Proliferation, Apoptosis |
| Address and Contact Information |
1 Department of Rheumatology and Immunology, Cangzhou Central Hospital, Cangzhou 061000, People’s Republic of China 2 The Second Nephrology Department, Cangzhou Central Hospital, Cangzhou 061000, People’s Republic of China * Corresponding author: liuliangcangzhou@163.com |
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| DOI: 10.1186/s11658-019-0152-2 Volume 24 (2019) - 24:28 | |
| Title | MiR-200c DOWNREGULATES HIF-1α AND INHIBITS MIGRATION OF LUNG CANCER CELLS |
| Authors | Yuree Byun†, Young-Chul Choi†, Yunhui Jeong, Gangtae Lee, Sena Yoon, Yongsu Jeong, Jaeseung Yoon and Kwanghee Baek* |
| Abstract | Background: Hypoxia-inducible factor-1α (HIF-1α) is a transcription factor with a
pivotal role in physiological and pathological responses to hypoxia. While HIF-1α is known to be involved in hypoxia-induced upregulation of microRNA (miRNA) expression, HIF-1α is also targeted by miRNAs. In this study, miRNAs targeting HIF-1α were identified and their effects on its expression and downstream target genes under hypoxic conditions were investigated. Cell migration under the same conditions was also assessed. Methods: microRNAs that target HIF-1α were screened using 3′-untranslated region luciferase (3′-UTR-luciferase) reporter assays. The expression levels of HIF-1α and its downstream target genes after transfection with miRNA were assessed using quantitative RT-PCR and western blot analyses. The effect of the miRNAs on the transcriptional activity of HIF-1α was determined using hypoxia-responsive element luciferase (HRE-luciferase) assays. Cell migration under hypoxia was examined using the wound-healing assay. Results: Several of the 19 screened miRNAs considerably decreased the luciferase activity. Transfection with miR-200c had substantial impact on the expression level and transcription activity of HIF-1α. The mRNA level of HIF-1α downstream genes decreased in response to miR-200c overexpression. MiR-200c inhibited cell migration in normoxia and, to a greater extent, in hypoxia. These effects were partly reversed by HIF-1α expression under hypoxic conditions. Conclusion: miR-200c negatively affects hypoxia-induced responses by downregulating HIF-1α, a key regulator of hypoxia. Therefore, overexpression of miR-200c might have therapeutic potential as an anticancer agent that inhibits tumor hypoxia. |
| Keywords | microRNA, miR-200c, HIF-1α, Hypoxia |
| Address and Contact Information | Graduate School of Biotechnology, Kyung Hee University, Yongin, Republic of Korea * Corresponding author: khbaek@khu.ac.kr †Yuree Byun and Young-Chul Choi contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0156-y Volume 24 (2019) - 24:29 | |
| Title | C-Cbl NEGATIVELY REGULATES TRAF6-MEDIATED NF-κB ACTIVATION BY PROMOTING K48-LINKED POLYUBIQUITINATION OF TRAF6 |
| Authors | Hyun-Duk Jang1,2,3, Hye Zin Hwang4, Hyo-Soo Kim1,2,3,5 and Soo Young Lee6* |
| Abstract | Background: In its RING domain, tumor necrosis factor receptor-associated factor 6 (TRAF6) has ubiquitin E3 ligase activity that facilitates the formation of lysine 63-linked polyubiquitin chains. This activity is required to activate nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB) and plays an important role in the IκB kinase (IKK) complex. Methods: An in vitro ubiquitination assay was used to establish whether c-Cbl could promote TRAF6 ubiquitination. We assessed direct binding and performed fine mapping between c-Cbl and TRAF6 based on the results of an immunoprecipitation assay with cultured 293 T cells. The luciferase reporter assay was applied to establish if c-Cbl-mediated ubiquitination affected NF-κB activation after stimulus from various TRAF-mediated signals: tumor necrosis factor-α (TNF-α), receptor activator of NF-κB ligand (RANKL), and interleukin-1β (IL-1β). An in vivo ubiquitination assay was performed using endogenous immunoprecipitation of TRAF6 in bone marrow macrophages (BMMs) and osteoclasts. Results: Here, we report on a form of TRAF6 ubiquitination that is mediated by c-Cbl, leading to the formation of lysine 48-linked polyubiquitin chains. The NF-κB activity induced by RANKL and IL-1β treatment is inhibited when c-Cbl is overexpressed, while the NF-κB activity induced by TNFα treatment is not. c-Cbl inhibits NF-κB activity mediated by TRAF6, but not by TRAF2. These findings show that c-Cbl ubiquitin ligase activity is essential for TRAF6 ubiquitination and negative regulation of NF-κB activity. Fine mapping revealed that the proline-rich domain of c-Cbl is critical for interaction with TRAF6. Stimulation with RANKL or interferon-γ (IFN-γ) caused c-Cbl to bind to polyubiquitinated TRAF6. Conclusions: These findings indicate that the interaction of TRAF6 with c-Cbl causes lysine 48-linked polyubiquitination for both negative feedback regulation and signaling cross-talk between RANKL and IFN-γ. |
| Keywords | Tumor necrosis factor receptor-associated factor 6, Ubiquitin, E3 ligase, C-Cbl |
| Address and Contact Information |
1 National Leading Laboratory for Stem Cell Research, Seoul National University College of Medicine, Seoul, South Korea. 2 Korea Research-Driven Hospital, Biomedical Research Institute, Seoul National University Hospital, Seoul, South Korea. 3 Strategic Center of Cell & Bio Therapy, Seoul National University Hospital, Seoul, South Korea. 4 Department of Biotechnology, The Catholic University of Korea, Bucheon, South Korea. 5 Cardiovascular Center & Department of Internal Medicine, Seoul National University Hospital, Seoul, South Korea. 6 Department of Life Science and the Research Center for Cellular Homeostasis, Ewha Woman’s University, Seoul, South Korea * Corresponding author: leesy@ewha.ac.kr |
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| DOI: 10.1186/s11658-019-0153-1 Volume 24 (2019) - 24:30 | |
| Title | CONCENTRATION CHANGES IN GEMCITABINE AND ITS METABOLITES AFTER HYPERTHERMIA IN PANCREATIC CANCER CELLS ASSESSED USING RP-HPLC |
| Authors | HB Jin, L Lu, L Xie, JF Yang, XF Zhang and SL Ma* |
| Abstract | Background: Gemcitabine (2′,2′-difluoro-2′-deoxycytidine;dFdC) is a first-line chemotherapy drug for pancreatic cancer. Recently, a synergistic anti-tumor treatment of dFdC and hyperthermia has achieved good clinical results, but there are few reports on the molecular mechanism influenced by hyperthermia. This study is an initial exploration of the effects of hyperthermia on changes in the concentration of dFdC and its metabolites in pancreatic cancer cells. The aim is to provide a theoretical basis for clinical detection and pharmacokinetic research. Methods: PANC-1 cells at logarithmic growth phase were used as the experimental object. The MTT assay was performed to determine the half maximal inhibitory concentration (IC50) of dFdC. After PANC-1 cells were cultured in DMEM medium containing IC50dFdC and treated with hyperthermia at 41 °C or 43 °C, changes in the concentration of dFdC, 2′,2′-difluorodeoxyuridine (dFdU) and difluorodeoxycytidine triphosphate (dFdCTP) in the cells were tested using an optimized reverse phase high-performance liquid chromatography (RP-HPLC) protocol. Results: We found that 41°C and 43°C hyperthermia gave rise to a decrease in dFdC and dFdU content. At 41°C, the levels respectively fell to 9.28 and 30.93% of the baseline, and at 43°C, to 24.76 and 57.80%, respectively. The dFdCTP content increased by 21.82% at 41°C and 42.42% at 43°C. Conclusion: The two heat treatments could alter the mechanism of dFdC metabolism in PANC-1 cells. The effect of 43 °C hyperthermia is more significant. Our observations may be instrumental to explaining the higher anti-tumor efficacy of this combination therapy. |
| Keywords | Pancreatic cancer, Gemcitabine, Metabolite, Hyperthermia, RP-HPLC |
| Address and Contact Information | Department of Gastroenterology, Affiliated Hangzhou First People’s Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China * Corresponding author: shenglinma2018@163.com |
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| DOI: 10.1186/s11658-019-0155-z Volume 24 (2019) - 24:31 | |
| Title | MiR-107 FUNCTION AS A TUMOR SUPPRESSOR GENE IN COLORECTAL CANCER BY TARGETING TRANSFERRIN RECEPTOR 1 |
| Authors | Yuxiang Fu, Liewen Lin and Ligang Xia* |
| Abstract | Background: While microRNAs (miRNAs) are known to play a critical role in the progression of colorectal cancer, the role of miR-107 remains unknown. We evaluated its role and explored the underlying mechanism. Materials & methods: MTT, wound-healing, transwell migration and transwell invasion assays were performed to evaluate the role of miR-107 in SW629 cell proliferation, migration and invasion. Real time-PCR and dual-luciferase reporter gene, TFR1 overexpression and western blotting assays were used to explore the underlying mechanism. Results: MiR-107 is downregulated in colorectal cancer tissues and several human colorectal cancer cell lines. Low miR-107 expression often indicates a poor survival rate for colorectal cancer patients. MiR-107 suppresses the proliferation, migration and invasion of SW620 cells by negatively regulating transferrin receptor 1 (TFR1). Conclusion: MiR-107 suppresses the metastasis of colorectal cancer and could be a potential therapy target in colorectal cancer patients. |
| Keywords | Colorectal cancer, microRNA107 (miR-107), Cancer progress, Transferrin receptor 1 (TFR1) |
| Address and Contact Information | Department of Gastrointestinal Surgery, Shenzhen People’s Hospital, The Second Clinical Medical College of Jinan University, The First Affiliated Hospital of South University of Science and Technology, Shenzhen 518055, China * Corresponding author: doctorw2016@163.com |
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| DOI: 10.1186/s11658-019-0157-x Volume 24 (2019) - 24:32 | |
| Title | CURCUMIN SUPPRESSES EPITHELIAL-TO-MESENCHYMAL TRANSITION OF PERITONEAL MESOTHELIAL CELLS (HMrSV5) THROUGH REGULATION OF TRANSFORMING GROWTH FACTOR- ACTIVATED KINASE 1 (TAK1) |
| Authors | Jun-Li Zhao1*, Mei-Zi Guo2, Jun-Jun Zhu1, Ting Zhang1 and Dan-Yan Min1 |
| Abstract | Objective: Peritoneal fibrosis remains a serious complication of long-term peritoneal dialysis (PD) leading to peritoneal membrane ultrafiltration failure. Epithelial–mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs) is a key process of peritoneal fibrosis. Curcumin has been previously shown to inhibit EMT of renal tubular epithelial cells and prevent renal fibrosis. There are only limited reports on inhibition of PMCs-EMT by curcumin. This study aimed to investigate the effect of curcumin on the regulation of EMT and related pathway in PMCs treated with glucose-based PD. Methods: EMT of human peritoneal mesothelial cells (HMrSV5) was induced with glucose-based peritoneal dialysis solutions (PDS). Cells were divided into a control group, PDS group, and PDS group receiving varied concentrations of curcumin. Cell Counting Kit-8 (CCK-8) assay was used to measure cell viability, and a transwell migration assay was used to verify the capacity of curcumin to inhibit EMT in HMrSV5 cells. Real-time quantitative PCR and western blot were used to detect the expression of genes and proteins associated with the EMT. Results: High glucose PDS decreased cell viability and increased migratory capacity. Curcumin reversed growth inhibition and migration capability of human peritoneal mesothelial cells (HPMCs). In HMrSV5 cells, high glucose PDS also decreased expression of epithelial markers, and increased expression of mesenchymal markers, a characteristic of EMT. Real-time RT-PCR and western blot revealed that, compared to the 4.25% Dianeal treated cells, curcumin treatment resulted in increased expression of E-cadherin (epithelial marker), and decreased expression of α-SMA (mesenchymal markers) (P < 0.05). Furthermore, curcumin reduced mRNA expression of two extracellular matrix protein, collagen I and fibronectin. Curcumin also reduced TGF-β1 mRNA and supernatant TGF-β1 protein content in the PDS-treated HMrSV5 cells (P < 0.05). Furthermore, it significantly reduced protein expression of p-TAK1, p-JNK and p-p38 in PDS-treated HMrSV5 cells. Conclusions: Our results demonstrate that curcumin showed an obvious protective effect on PDS-induced EMT of HMrSV5 cells and suggest implication of the TAK1, p38 and JNK pathway in mediating the effects of curcumin in EMT of MCs. |
| Keywords | Peritoneal fibrosis (PF), Peritoneal dialysis, Epithelial-mesenchymal transition (EMT), Curcumin, TGF-β-activated kinase 1 (TAK1) |
| Address and Contact Information |
1 Department of Nephrology, Shanghai University of Medicine & Health Sciences affiliated Zhoupu Hospital, Pudong New District, Shanghai 201318, China 2 Department of Geriatrics, Shanghai University of Medicine & Health Sciences affiliated Zhoupu Hospital, Pudong New District, Shanghai 201318, China. * Corresponding author: zhaojunli1203@126.com |
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| DOI: 10.1186/s11658-019-0159-8 Volume 24 (2019) - 24:33 | |
| Title | NUTRIENT DEPRIVATION AND LYSOSOMAL STRESS INDUCE ACTIVATION OF TFEB IN RETINAL PIGMENT EPITHELIAL CELLS |
| Authors | Hsuan-Yeh Pan1, Abdulla H. Alamri2 and Mallika Valapala1* |
| Abstract | Background: Induction of lysosomal function and autophagy is regarded as an adaptive mechanism in response to cellular stress. The transcription factor EB (TFEB) has been identified as a master regulator of lysosomal function and autophagy. TFEB is a member of the microphthalmia family of bHLH-LZ transcription factors that includes other members such as micropthalmia-associated transcription factor (MITF), TFE3, and TFEC. TFEB controls lysosome biogenesis and autophagy by upregulation of a family of genes belonging to the Coordinated Lysosomal Expression and Regulation (CLEAR) network. Here, we investigated the expression of TFEB in cells subjected to nutrient deprivation and lysosomal stress. We studied transcriptional induction of TFEB-regulated genes in response to nutrient deprivation and lysosomal stress in retinal pigment epithelial (RPE) cells. Furthermore, we also investigated the induction of autophagy and lysosomal genes upon overexpression of constitutively active form of TFEB. Methods: Expression of TFEB and MITF protein levels were evaluated in cells subjected to prolonged periods of nutrient deprivation. mRNA levels of the CLEAR network genes was measured by quantitative real time PCR (qRT-PCR) analysis in cells deprived of nutrients, treated with ammonium chloride and upon overexpression of constitutively active TFEB. Immunostaining with LC3 antibody was used to measure autophagy flux. Labeling with lysoTracker dye was used to assess lysosomes. Results: Our results show that nutrient deprivation increases protein levels of TFEB and MITF in ARPE-19 cells. Nutrient stress induces the expression of lysosomal (LAMP1, CTSD MCOLN1, SGSH) and autophagy (BECN1) genes. Lysosomal stress also increases the expression of lysosomal (ATP6V0A1 and LAMP1) and autophagy (p62 and BECN1) genes. Our results show that overexpression of constitutively active TFEB also induces the expression of CLEAR network genes. Conclusions: Collectively, these observations suggest that nutrient stress induces the protein expression of both MITF and TFEB in ARPE-19 cells. TFEB-regulated transcriptional program plays an important role in adaptive response of cells during both nutrient and lysosomal stress. |
| Keywords | Nutrient deprivation, Lysosomal stress, Autophagy |
| Address and Contact Information |
1 School of Optometry, Indiana University, Bloomington, IN 47405, USA 2 State University of New York College of Optometry, 33 42nd St., New York, NY 10036, USA. * Corresponding author: mvalapal@iu.edu |
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| DOI: 10.1186/s11658-019-0161-1 Volume 24 (2019) - 24:34 | |
| Title | MicroRNA-340-5p SUPPRESSES NON-SMALL CELL LUNG CANCER CELL GROWTH AND METASTASIS BY TARGETING ZNF503 |
| Authors | Guojie Lu and Yaosen Zhang* |
| Abstract | Background: MicroRNAs (miRNAs) have been reported to play crucial roles in cancer cell processes, including proliferation, metastasis and cell cycle progression. We aimed to identify miRNAs that could act as suppressors of cell growth and invasion in non-small cell lung cancer (NSCLC). Methods: Fifteen paired NSCLC tissue samples and pericarcinomatous normal tissues were collected and preserved in liquid nitrogen. The expression levels of miR-340-5p and ZNF503 mRNA were detected using a qPCR assay. The transfection of plasmids was conducted using Lipofectamine 3000 according to the manufacturer’s protocol. Cell proliferation was determined using a CCK-8 assay. The protein levels of endothelial–mesenchymal transition markers were measured using a western blot assay. Cell invasive ability was evaluated using a transwell assay. TargetScan was used to predict targets of miR-340. A dual luciferase reporter assay was performed to confirm a potential direct interaction between miR-340-5p and ZNF503. Results: The expression level of miR-340-5p was frequently found to be lower in NSCLC tissues than in matched pericarcinomatous normal tissues. Overexpression of miR-340-5p significantly inhibited the proliferation and invasion NCI-H1650 (a NSCLC cell line), while inhibition of miR-340-5p stimulated cell growth. Using TargetScan, we predicted that ZNF503 could be a target of miR-340-5p. Further mechanistic studies demonstrated that the forced expression of ZNF503 could partially abrogate the miR-340-5p-mediated decrease in NCI-H1650 cell viability and invasion, suggesting that miR-340-5p suppressed cell growth and invasion in a ZNF503-dependent manner. Conclusion: Our findings indicate that miR-340-5p inhibits NCI-H1650 cell proliferation and invasion by directly targeting ZNF503 and that miR-340-5p can serve as a potential therapeutic target for treating NSCLC. |
| Keywords | Cell proliferation, Metastasis, miR-340-5p, NSCLC, ZNF503 |
| Address and Contact Information | Department of Thoracic Surgery, Guangzhou Panyu District Central Hospital, No. 8, Fuyu East Road, Qiaonan Street, Panyu District, Guangzhou 511486, People’s Republic of China * Corresponding author: 13632332051@163.com |
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| DOI: 10.1186/s11658-019-0154-0 Volume 24 (2019) - 24:35 | |
| Title | ROSIGLITAZONE PROMOTES ENaC-MEDIATED ALVEOLAR FLUID CLEARANCE IN ACUTE LUNG INJURY THROUGH THE PPARγ/SGK1 SIGNALING PATHWAY |
| Authors | Jing He, Di Qi, Xu-mao Tang, Wang Deng, Xin-yu Deng, Yan Zhao* and Dao-xin Wang* |
| Abstract | Background: Pulmonary edema is one of the pathological characteristics of acute respiratory distress syndrome (ARDS). The epithelial sodium channel (ENaC) is thought to be the rate-limiting factor for alveolar fluid clearance (AFC) during pulmonary edema. The peroxisome proliferator-activated receptor γ (PPARγ) agonist rosiglitazone was shown to stimulate ENaC-mediated salt absorption in the kidney. However, its role in the lung remains unclear. Here, we investigated the role of the PPARγ agonist in the lung to find out whether it can regulate AFC during acute lung injury (ALI). We also attempted to elucidate the mechanism for this. Methods: Our ALI model was established through intratracheal instillation of lipopolysaccharide (LPS) in C57BL/6 J mice. The mice were randomly divided into 4 groups of 10. The control group underwent a sham operation and received an equal quantity of saline. The three experimental groups underwent intratracheal instillation of 5 mg/kg LPS, followed by intraperitoneal injection of 4 mg/kg rosiglitazone, 4 mg/kg rosiglitazone plus 1 mg/kg GW9662, or only equal quantity of saline. The histological morphology of the lung, the levels of TNF-α and IL-1β in the bronchoalveolar lavage fluid (BALF), the level of AFC, and the expressions of αENaC and serum and glucocorticoid-induced kinase-1 (SGK1) were determined. Type 2 alveolar (AT II) cells were incubated with rosiglitazone (15 μM) with or without GW9662 (10 μM). The expressions of αENaC and SGK1 were determined 24 h later. Results: A mouse model of ALI was successfully established. Rosiglitazone significantly ameliorated the lung injury, decreasing the TNF-α and IL-1β levels in the BALF, enhancing AFC, and promoting the expressions of αENaC and SGK1 in ALI mice, which were abolished by the specific PPARγ blocker GW9662. In vitro, rosiglitazone increased the expressions of αENaC and SGK1. This increase was prevented by GW9662. Conclusions: Rosiglitazone ameliorated the lung injury and promoted ENaC-mediated AFC via a PPARγ/SGK1-dependent signaling pathway, alleviating pulmonary edema in a mouse model of ALI. |
| Keywords | Acute lung injury, Alveolar epithelial sodium channel, Alveolar fluid clearance, Peroxisome proliferator-activated receptor γ, Serum and glucocorticoid induced kinase-1 |
| Address and Contact Information | Department of Respiratory Medicine, The Second Affiliated Hospital of Chongqing Medical University, 76 Linjiang Road, Yuzhong District, Chongqing 400010, China * Corresponding author: 514342948@qq.com; submitmail@126.com |
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| DOI: 10.1186/s11658-019-0158-9 Volume 24 (2019) - 24:36 | |
| Title | THE ROLE OF DIFFERENT SIRT1-MEDIATED SIGNALING PATHWAYS IN TOXIC INJURY |
| Authors | Zhihua Ren1†, Hongyi He1†, Zhicai Zuo1†, Zhiwen Xu1, Zhanyong Wei2* and Junliang Deng1* |
| Abstract | Common environmental pollutants and drugs encountered in everyday life can cause toxic damage to the body through oxidative stress, inflammatory stimulation, induction of apoptosis, and inhibition of energy metabolism. Silent information regulator 1 (SIRT1), a nicotinamide adenine dinucleotide-dependent deacetylase, is a member of the evolutionarily highly conserved Sir2 (silent information regulator 2) superprotein family, which is located in the nucleus and cytoplasm. It can deacetylate protein substrates in various signal transduction pathways to regulate gene expression, cell apoptosis and senescence, participate in the process of neuroprotection, energy metabolism, inflammation and the oxidative stress response in living organisms, and plays an important role in toxic damage caused by toxicants and in the process of SIRT1 activator/inhibitor antagonized toxic damage. This review summarizes the role that SIRT1 plays in toxic damage caused by toxicants via its interactions with protein substrates in certain signaling pathways. |
| Keywords | SIRT1, Toxicant, SIRT1 activator, Signaling pathway |
| Address and Contact Information |
1 Key Laboratory of Animal Disease and Human Health of Sichuan Province, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, Sichuan Province, China 2 The College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, Henan Province, China * Corresponding author: weizhanyong@henau.edu.cn; dengjl213@126.com † Zhihua Ren, Hongyi He and Zhicai Zuo contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0160-2 Volume 24 (2019) - 24:37 | |
| Title | INHIBITION OF miR-19a PROTECTS NEURONS AGAINST ISCHEMIC STROKE THROUGH MODULATING GLUCOSE METABOLISM AND NEURONAL APOPTOSIS |
| Authors | Xiao-Li Ge1, Jin-Li Wang1, Xin Liu2, Jia Zhang3, Chang Liu4 and Li Guo1* |
| Abstract | Background: Accumulating evidence has shown that altered microRNA (miR) modulation is implicated in the pathologies of ischemic stroke. However, it is unclear whether and how hsa-miR-19a-3p mediates cerebral ischemic injury. Herein, we investigated the functional role of miR-19a-3p in cerebral ischemic injury and explored its underlying regulatory mechanism. Methods: In vivo ischemic/reperfusion (I/R) neuronal injury and in vitro oxygen-glucose deprivation (OGD) were established. Expression of miR-19a-3p was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Glucose uptake, lactate production, and apoptosis were determined. ADIPOR2 was predicted as a target of miR-19a-3p in silico and experimentally validated by qRT-PCR, Western blot analysis and luciferase assay assays. Results: MiR-19a expression was significantly downregulated and upregulated in rat neurons and astrocytes, respectively (P < 0.01). A significantly elevated level of miR-19a-3p was found in I/R and OGD models in comparison to sham/control groups (P < 0.01). Expression of the glycolysis enzyme markers LDHA, PKM2, HK2, Glut1 and PDK1, apoptosis-related factors levels, apoptosis, glucose uptake, and lactate production were significantly repressed by both I/R and OGD (P < 0.01 in each case). Moreover, miR-19a-3p mimic aggravated, while miR-19a-3p inhibitor alleviated, the above observations. Adipor2 was predicted and confirmed to be a direct target of miR-19a. Furthermore, restoration of Adipor2 reversed miR-19a-3p-induced effects. Conclusions: Collectively, our results indicate that elevated miR-19a-3p mediates cerebral ischemic injury by targeting ADIPOR2. MiR-19a-3p attenuation thus might offer hope of a novel therapeutic target for ischemic stroke injury treatment. |
| Keywords | Ischemic stroke, miR-19a-3p, ADIPOR2, Glucose metabolism, Apoptosis |
| Address and Contact Information |
1 Department of Neurology, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, China 2 Department of Neurosurgery, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, China. 3 Department of Obstetrics, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, China. 4 Department of Rehabilitation, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, China. * Corresponding author: guoli_med@126.com |
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| DOI: 10.1186/s11658-019-0162-0 Volume 24 (2019) - 24:38 | |
| Title | TRANSCRIPTOME PROFILING REVEALED MULTIPLE GENES AND ECM-RECEPTOR INTERACTION PATHWAYS THAT MAY BE ASSOCIATED WITH BREAST CANCER |
| Authors | Yulong Bao1,3†, Li Wang1†, Lin Shi2, Fen Yun2, Xia Liu2, Yongxia Chen3, Chen Chen2, Yanni Ren2 and Yongfeng Jia1,3* |
| Abstract | Background: Exploration of the genes with abnormal expression during the development of breast cancer is essential to provide a deeper understanding of the mechanisms involved. Transcriptome sequencing and bioinformatics analysis of invasive ductal carcinoma and paracancerous tissues from the same patient were performed to identify the key genes and signaling pathways related to breast cancer development. Methods: Samples of breast tumor tissue and paracancerous breast tissue were obtained from 6 patients. Sequencing used the Illumina HiSeq platform. All. Only perfectly matched clean reads were mapped to the reference genome database, further analyzed and annotated based on the reference genome information. Differentially expressed genes (DEGs) were identified using the DESeq R package (1.10.1) and DEGSeq R package (1.12.0). Using KOBAS software to execute the KEGG bioinformatics analyses, enriched signaling pathways of DEGs involved in the occurrence of breast cancer were determined. Subsequently, quantitative real time PCR was used to verify the accuracy of the expression profile of key DEGs from the RNA-seq result and to explore the expression patterns of novel cancer-related genes on 8 different clinical individuals. Results: The transcriptomic sequencing results showed 937 DEGs, including 487 upregulated and 450 downregulated genes in the breast cancer specimens. Further quantitative gene expression analysis was performed and captured 252 DEGs (201 downregulated and 51 upregulated) that showed the same differential expression pattern in all libraries. Finally, 6 upregulated DEGs (CST2, DRP2, CLEC5A, SCD, KIAA1211, DTL) and 6 downregulated DEGs (STAC2, BTNL9, CA4, CD300LG, GPIHBP1 and PIGR), were confirmed in a quantitative real time PCR comparison of breast cancer and paracancerous breast tissues from 8 clinical specimens. KEGG analysis revealed various pathway changes, including 20 upregulated and 21 downregulated gene enrichment pathways. The extracellular matrix–receptor (ECM-receptor) interaction pathway was the most enriched pathway: all genes in this pathway were DEGs, including the THBS family, collagen and fibronectin. These DEGs and the ECM-receptor interaction pathway may perform important roles in breast cancer. Conclusion: Several potential breast cancer-related genes and pathways were captured, including 7 novel upregulated genes and 76 novel downregulated genes that were not found in other studies. These genes are related to cell proliferation, movement and adhesion. They may be important for research into breast cancer mechanisms, particularly CST2 and CA4. A key signaling pathway, the ECM-receptor interaction signal pathway, was also identified as possibly involved in the development of breast cancer. |
| Keywords | Breast cancer, Transcriptome, Differentially expressed genes, ECM-receptor interaction pathway |
| Address and Contact Information |
1 College of Basic Medicine, Inner Mongolia Medical University, Hohhot, Inner Mongolia, China 2 Department of Pathology, Inner Mongolia Medical University, Hohhot, Inner Mongolia, China. 3 Tumor Molecular Diagnostic Laboratory, The Inner Mongolia Cancer Hospital, Hohhot, Inner Mongolia, China * Corresponding author: jiayf0471@163.com †Yulong Bao and Li Wang contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0163-z Volume 24 (2019) - 24:39 | |
| Title | NON-CONTACT CO-CULTURE WITH HUMAN VASCULAR ENDOTHELIAL CELLS PROMOTES EPITHELIAL-TO-MESENCHYMAL TRANSITION OF CERVICAL CANCER SIHA CELLS BY ACTIVATING THE NOTCH1/LOX/SNAIL PATHWAY |
| Authors | Jinghua Ou1,2, Defeng Guan1 and Yongxiu Yang1,3* |
| Abstract | Background: The aim of this study was to investigate the effect of human umbilical vein endothelial cells on epithelial-to-mesenchymal transition of the cervical cancer cell line SiHa by studying the Notch1/lysyl oxidase (LOX)/SNAIL1 pathway. Methods: Monocultures of SiHa cells, SiHa cells containing a control sequence, and Notch1-silenced SiHa cells, as well as co-cultures of human umbilical vein endothelial cells with SiHa cells and Notch1-silenced SiHa cells, were established. The invasiveness of SiHa cells in each group was evaluated using a Transwell assay. The mRNA levels of E-cadherin and vimentin were detected using quantitative real-time polymerase chain reaction. The expression levels of the matrix metalloproteinases MMP-2 and MMP-9 were determined in SiHa cells using an immunofluorescence assay and the protein activity was detected by gelatin zymography. Changes in LOX, SNAIL1 and NOTCH1 expression in the SiHa cells in each group were detected using western blotting. Results: Compared with monocultured SiHa cells, co-cultured SiHa cells showed a significant increase in their invasiveness and expression levels of vimentin, as well as of NOTCH 1, LOX, and SNAIL1, whereas their expression of E-cadherin was significantly reduced and protein activities of MMP-2 and MMP-9 were increased. Compared with SiHa, mono- and co-cultured NOTCH 1-silenced SiHa cells showed significant reductions in their invasiveness and expression levels of vimentin, NOTCH 1, LOX, and SNAIL1, whereas their expression of E-cadherin significantly increased and protein activities of MMP-2 and MMP-9 decreased. Conclusion: Co-culture with human umbilical vein endothelial cells promoted the epithelial-to-mesenchymal transition of SiHa cells by activating the NOTCH1/LOX/SNAIL1 pathway in SiHa cells, which enhanced their invasive and metastatic capacities. The results of this study may provide a new perspective on cervical cancer metastasis and a theoretical basis for clinical treatment. |
| Keywords | Co-culture, Cervical cancer, SiHa cells, HUVECs, NOTCH/LOX/SNAIL pathway, EMT |
| Address and Contact Information |
1 The First Clinical Medical College of Lanzhou University, NO 1 West Donggang Road, Chengguan District, Lanzhou, Gansu, China. 2 Department of obstetrics and gynecology, Gansu provincial hospital, Lanzhou, Gansu, China. 3 Department of Obstetrics and Gynecology, The First Hospital of Lanzhou University, NO 1 West Donggang Road, Chengguan District, Lanzhou, Gansu, China. * Corresponding author: yyxfhlz@163.com |
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| DOI: 10.1186/s11658-019-0164-y Volume 24 (2019) - 24:40 | |
| Title | PROGRESS IN RESEARCH ON PACLITAXEL AND TUMOR IMMUNOTHERAPY |
| Authors | Linyan Zhu and Liqun Chen* |
| Abstract | Paclitaxel is a well-known anticancer agent with a unique mechanism of action. It is considered to be one of the most successful natural anticancer drugs available. This study summarizes the recent advances in our understanding of the sources, the anticancer mechanism, and the biosynthetic pathway of paclitaxel. With the advancement of biotechnology, improvements in endophytic fungal strains, and the use of recombination techniques and microbial fermentation engineering, the yield of extracted paclitaxel has increased significantly. Recently, paclitaxel has been found to play a large role in tumor immunity, and it has a great potential for use in many cancer treatments. |
| Keywords | Paclitaxel, Anticancer mechanism, Endophytic fungus, Biosynthetic pathway |
| Address and Contact Information | College of Biological Science and Engineering, Fuzhou University, Fuzhou 350108, China * Corresponding author: lqchen@fzu.edu.cn |
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| DOI: 10.1186/s11658-019-0165-x Volume 24 (2019) - 24:41 | |
| Title | LncRNA GASL1 IS DOWNREGULATED IN CHRONIC HEART FAILURE AND REGULATES CARDIOMYOCYTE APOPTOSIS |
| Authors | Haihong Deng1, Wenbo Ouyang1, Li Zhang3, Xiaoshan Xiao4, Zhiyong Huang3* and Wendian Zhu2* |
| Abstract | Background: TGF-β1 contributes to chronic heart failure. It is known that lncRNA GASL1 can inactivate TGF-β1 in cancer biology. Methods: All the participants were enrolled in the First People’s Hospital of Zhaoqing during the period June 2012 to June 2013. ELISA, RT-qPCR, vectors, transient transfections and western blot were carried out during the research. Results: We found that plasma levels of TGF-β1 were significantly higher, while levels of GASL1 in plasma were significantly lower in chronic heart failure (CHF) patients compared to the control group. TGF-β1 and GASL1 were inversely correlated in CHF patients. Low pretreatment plasma levels of GASL1 were closely associated with poor survival of CHF patients. GASL1 expression was not significantly affected by TGF-β1 overexpression in cardiomyocytes, while cardiomyocytes with GASL1 overexpression showed downregulated TGF-β1. Overexpression of GASL1 led to a decreased, while TGF-β1 overexpression led to an increased apoptotic rate of cardiomyocytes under H2O2 treatment. In addition, TGF-β1 overexpression attenuated the effect of GASL1 overexpression. Conclusion: In conclusion, GASL1 was downregulated in CHF. GASL1 overexpression may improve CHF by inhibiting cardiomyocyte apoptosis through the inactivation of TGF-β1. |
| Keywords | Chronic heart failure, lncRNA GASL1, TGF-β1, Apoptosis |
| Address and Contact Information |
1Department of Anesthesiology, The First People’s Hospital of Zhaoqing, Zhaoqing City, Guangdong Province 526000, People’s Republic of China. 2Department of General Surgery, The First People’s Hospital of Zhaoqing, No. 9 Donggang East Road, Duanzhou District, Zhaoqing City, Guangdong Province 526000, People’s Republic of China 3Department of Anesthesiology, Fuwai Hospital Chinese Academy of Medical Sciences, No. 12 Langshan Road, Shenzhen City 518057, People’s Republic of China * Corresponding author: huzhgy@gmail.com; jl59911@163.com |
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| DOI: 10.1186/s11658-019-0169-6 Volume 24 (2019) - 24:42 | |
| Title | Dp71 DEPLETED HBE CELLS DISPLAYED INCREASED DNA DAMAGE AND APOPTOSIS INDUCED BY H2O2 |
| Authors | Sichuang Tan2, Shuai Zhao2,6, Xuefei Xiao5, Lan Xiao4, Jinliang Xie3 and Sipin Tan1* |
| Abstract | Human bronchial epithelium (HBE)-Dp71 anti-sense(AS)cells with stably transfected Dp71 siRNA plasmids were prepared for further exploration of Dp71 biological traits in cells other than PC12. HBE-Dp71AS cells displayed increased DNA damage induced by H2O2. Apoptosis of HBE-Dp71AS cells induced by H2O2 was increased via enhancing caspase 3, caspase 8 and caspase 9. HBE-Dp71AS cells also displayed decreased proliferation and clonogenic formation. RAD51 was proved to be a new binding partner of Dp71 by co-immunoprecipitation (Ip) and immunofluorescence. Reduced RAD51 mRNA and protein levels were observed in HBE-Dp71AS cells. Decreased lamin B1, focal adhesion kinase (FAK), phosphorylated focal adhesion kinase (p-FAK) and phosphorylated protein kinase B (p-AKT) were detected in the HBE-Dp71AS cells, which functioned together with RAD51 as the molecular explanations for the character alterations of HBE-Dp71AS cells. |
| Keywords | Dp71, DNA damage, Apoptosis, RAD51, FAK |
| Address and Contact Information |
1 Key Laboratory of Sepsis Translational Medicine of Hunan, Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha, Hunan Province 410008, People’s Republic of China 2 Department of Thoracic Surgery, the Second Xiangya Hospital, Central South University, 139 Ren-min Road, Changsha, Hunan Province 410011, People’s Republic of China. 4 Department of Traditional Chinese Medicine, the Third Xiangya Hospital, Central South University, Changsha, Hunan Province, People’s Republic of China. 5 Department of Emergency and Critical Care Medicine, the Third Xiangya Hospital, Central South University, Changsha, Hunan Province, People’s Republic of China. 6 Department of Pathology, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, People’s Republic of China. * Corresponding author: springtan@csu.edu.cn |
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| DOI: 10.1186/s11658-019-0167-8 Volume 24 (2019) - 24:43 | |
| Title | MESENCHYMAL STEM CELLS DECREASE BLOOD–BRAIN BARRIER PERMEABILITY IN RATS WITH SEVERE ACUTE PANCREATITIS |
| Authors | Ronggui Lin1†, Ming Li2†, Meiqin Luo3, Tianhong Teng1, Yu Pan1 and Heguang Huang1* |
| Abstract | Background: Impairment of the blood–brain barrier (BBB) could result in secondary cerebral edema and life-threatening pancreatic encephalopathy in patients with severe acute pancreatitis (SAP). Mesenchymal stem cells (MSCs) have been widely adopted in clinical research because of their pleiotropic functions. The aim of this study was to investigate the impact of MSCs on BBB permeability in SAP and the potential mechanisms driving these effects. Methods: Sprague-Dawley rats were randomly assigned to the control, SAP and SAP+MSCs groups. Pancreatic impairment was assessed. The serum levels of amylase, TNF-α and IL-10, expression levels of claudin-5, Bax, Bcl-2 and MMP-9, and the BBB permeability were measured. Endothelial cell apoptosis was evaluated. Results: SAP rats showed BBB impairment with increased permeability and secondary cerebral edema, which was confirmed using the Evans blue assay and the calculation of the brain dry/wet ratio. Treatment with MSCs decreased the serum levels of amylase and TNF-α, increased the serum levels of IL-10, attenuated the apoptosis of brain microvascular endothelial cells, upregulated claudin-5 expression and downregulated MMP-9 expression. This treatment attenuated the increased BBB permeability in SAP rats. Conclusions: MSCs attenuated the impairment of the BBB and decreased its permeability, producing protective effects in SAP rats. |
| Keywords | Severe acute pancreatitis, Blood–brain barrier, Mesenchymal stem cell, Inflammatory response, Endothelial cell |
| Address and Contact Information |
1 Department of General surgery, Fujian Medical University Union Hospital, 29 Xinquan Road, Fuzhou, Fujian 350001, People’s Republic of China 2 Department of Histology and Embryology, Hunan University of Medicine, Huaihua, Hunan, China. 3 Department of Orthopedics, Fujian Medical University Union Hospital, Fuzhou, Fujian, China. * Corresponding author: Heguanghuang2@163.com † Ronggui Lin and Ming Li contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0171-z Volume 24 (2019) - 24:44 | |
| Title | THE EFFECTS OF CRISPR-Cas9 KNOCKOUT OF THE TGF-β1 GENE ON ANTLER CARTILAGE CELLS IN VITRO |
| Authors | Mingxiao Liu, Xiangyu Han, Hongyun Liu, Danyang Chen, Yue Li and Wei Hu* |
| Abstract | Background: Deer antler is the only mammalian organ that can be completely regenerated every year. Its periodic regeneration is regulated by multiple factors, including transforming growth factor β (TGF-β). This widely distributed multi-functional growth factor can control the proliferation and differentiation of many types of cell, and it may play a crucial regulatory role in antler regeneration. This study explored the role of TGF-β1 during the rapid growth of sika deer antler. Methods: Three CRISPR-Cas9 knockout vectors targeting the TGF-β1 gene of sika deer were constructed and packaged with a lentiviral system. The expression level of TGF-β1 protein in the knockout cell line was determined using western blot, the proliferation and migration of cartilage cells in vitro were respectively determined using EdU and the cell scratch test, and the expression levels of TGF-β pathway-related genes were determined using a PCR array. Results: Of the three gRNAs designed, pBOBI-gRNA2 had the best knockout effect. Knockout of TGF-β1 gene inhibits the proliferation of cartilage cells and enhances their migration in vitro. TGF-β signaling pathway-related genes undergo significant changes, so we speculate that when the TGF-β pathway is blocked, the BMP signaling pathway mediated by BMP4 may play a key role. Conclusions: TGF-β1 is a newly identified regulatory factor of rapid growth in sika deer antler. |
| Keywords | CRISPR-Cas9, Sika deer, Transforming growth factor-β1, Cartilage cells |
| Address and Contact Information | College of Life Sciences, Jilin Agriculture University, Changchun 130118, Jilin Province, China * Corresponding author: huweilab@126.com |
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| DOI: 10.1186/s11658-019-0170-0 Volume 24 (2019) - 24:45 | |
| Title | THE EFFECT OF AP-2δ ON TRANSCRIPTION OF THE PRESTIN GENE IN HEI-OC1 CELLS UPON OXIDATIVE STRESS |
| Authors | Xuan Luo1, Yun Xia2, Xu-Dong Li3 and Jun-Yi Wang1* |
| Abstract | Background: The study aimed to investigate the effect of oxidative stress on Prestin expression, and explore the transcription factors (TFs) that are involved in regulating the expression of Prestin in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells upon oxidative stress. Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression level of Prestin. Reverse chromatin immunoprecipitation (reverse ChIP) assay was performed to identify proteins that could bind to the Prestin gene. Small interfering RNA (siRNA) and chromatin immunoprecipitation (ChIP) experiments were used to further verify the results. HEI-OC1 cells were incubated with four different concentrations of tert-butyl hydroperoxide (t-BHP) for 24 h or 48 h to construct the oxidative stress model. Results: Oxidative stress induced Prestin increase at the mRNA level but with a concomitant decrease at the protein level. TF activating enhancer binding protein-2δ (AP-2δ) screened by reverse ChIP assay was demonstrated to bind to transcriptional start site 1441 of the Prestin promoter region and negatively regulate the expression of Prestin by siRNA and ChIP experiments. Furthermore, AP-2δ was down-regulated under oxidative stress. Conclusions: In conclusion, oxidative stress inhibits the expression of Prestin protein, and the transcription mechanism is triggered to compensate for the loss of Prestin protein. AP-2δ is one of the important TFs that suppresses transcription of the Prestin gene, and AP-2δ suppression further boosted Prestin mRNA activation under oxidative stress. |
| Keywords | Prestin, AP-2δ, Oxidative stress, Transcriptional regulation, HEI-OC1 cells |
| Address and Contact Information |
1 Department of Labor Health and Environmental Hygiene, School of Public Health, Guangdong Pharmaceutical University, Guangzhou 510310, China 2 Department of Labor Health and Environmental Hygiene, School of Public Health, Guangdong Pharmaceutical University, Guangzhou 510310, China. 3 Key Laboratory, Occupational Disease Prevention and Control of Hospital of Guangdong Province, Guangzhou 510300, China. * Corresponding author: wjy925@163.com |
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| DOI: 10.1186/s11658-019-0166-9 Volume 24 (2019) - 24:46 | |
| Title | MiR-216a-5p TARGETS TCTN1 TO INHIBIT CELL PROLIFERATION AND INDUCE APOPTOSIS IN ESOPHAGEAL SQUAMOUS CELL CARCINOMA |
| Authors | Lixun Chai and Gengpu Yang* |
| Abstract | Background: MiR-216a-5p has been reported to be associated with several tumors, including prostate cancer and melanoma. However, its expression level and potential role in esophageal squamous cell carcinoma (ESCC) remain uncertain. Results: Here, we found that miR-216a-5p expression was significantly down-regulated in clinical ESCC tissues and cells. Functional assays were performed to evaluate the biological effects of miR-216a-5p on cell proliferation and cell apoptosis by CCK-8 assay and flow cytometry in ESCC cell lines, EC9706 and TE-9. The results showed that miR-216a-5p overexpression repressed cell proliferation and induced cell apoptosis. Through bioinformatics prediction and luciferase reporter assay, we revealed that miR-216a-5p could directly target tectonic family member 1 (TCTN1). Moreover, TCTN1 was obviously suppressed by miR-216a-5p overexpression. In addition, TCTN1 expression was significantly increased and inversely correlated with the levels of miR-216a-5p in ESCC tissues. More importantly, down-regulation of TCTN1 imitated, while restoration of TCTN reversed the effects of miR-216a-5p on cell proliferation and apoptosis. At the molecular level, we further found that TCTN1 overexpression reversed the effects of miR-216a-5p transfection on the expression of PCNA, Bcl-2 and Bad. Conclusions: Our results demonstrate that miR-216a-5p might serve as a tumor suppressor in ESCC cells through negatively regulating TCTN1 expression, indicating the possibility that miR-216a-5p and TCTN1 might be attractive targets for ESCC therapeutic intervention. |
| Keywords | ESCC, miR-216a-5p, TCTN1, Proliferation, Apoptosis |
| Address and Contact Information | Department of Thoracic Surgery, Shanxi Dayi Hospital, No. 99 Dragon City Street, Taiyuan 030012, Shanxi Province, China * Corresponding author: yang_Gpu@163.com |
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| DOI: 10.1186/s11658-019-0168-7 Volume 24 (2019) - 24:47 | |
| Title | MicroRNA-5195-3p ENHANCES THE CHEMOSENSITIVITY OF TRIPLE-NEGATIVE BREAST CANCER TO PACLITAXEL BY DOWNREGULATING EIF4A2 |
| Authors | Mei Liu, Can Gong, Renyuan Xu, Yu Chen and Xiaodong Wang* |
| Abstract | Background: Chemotherapy based on paclitaxel (PTX) is the standard treatment for a range of cancers, including triple-negative breast cancer (TNBC), but the increasing development of resistance has reduced/has negatively impacted its clinical utility. A previous study demonstrated that miR-5195-3p could suppress lung cancer cell growth. This study was designed to investigate whether miR-5195-3p attenuates chemoresistance to PTX by regulating target genes in TNBC cells. Methods: The study used both PTX-resistant tumor tissues and PTX-resistant TNBC cell lines. The expression of miR-5195-3p was determined using quantitative real-time PCR. Cell viability, cell cycle distribution and apoptosis were analyzed using CCK-8 and flow cytometry assays. The target genes of miR-5195-3p were predicted with bioinformatics analysis and confirmed using the luciferase reporter assay. Results: MiR-5195-3p expression was lower in PTX-resistant tumor tissues and PTX-resistant TNBC cell lines. Upregulation of miR-5195-3p enhanced the sensitivity of PTX-resistant TNBC cells to PTX treatment. EIF4A2 was confirmed as a potential target of miR-5195-3p. EIF4A2 knockdown imitated the effects of miR-5195-3p on chemosensitivity, while restoration of EIF4A2 rescued them. Conclusion: These data demonstrate that miR-5195-3p might be a potential therapeutic target to reverse chemoresistance in TNBC through its targeting of EIF4A2. |
| Keywords | Triple-negative breast cancer, miR-5195-3p, EIF4A2, Chemosensitivity |
| Address and Contact Information | Department of Breast Surgery, West China Hospital of Sichuan University, 37 Guoxue Lane, Wuhou District, Chengdu 610041, Sichuan Province, China * Corresponding author: wang_xdong2018@126.com |
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| DOI: 10.1186/s11658-019-0173-x Volume 24 (2019) - 24:48 | |
| Title | microRNA-211 REGULATES CELL PROLIFERATION, APOPTOSIS AND MIGRATION/INVASION IN HUMAN OSTEOSARCOMA VIA TARGETING EZRIN |
| Authors | Yihua Pei1,2†, Qin Yao1,2†, Yingchao Li3, Xin Zhang4 and Bozhen Xie3* |
| Abstract | Background: In recent years, microRNA-211 (miR211) has been considered as a tumor suppressor in multiple malignancies. However, the function of miR211 in human osteosarcoma has not been explored intensively so far. In this study, the relationship between miR211 and EZRIN was analyzed in human osteosarcoma. Methods: The expression levels of miR211 and EZRIN were measured in both human osteosarcoma cells and tissues. The direct regulatory relationship between miR211 and EZRIN was evaluated using dual-luciferase assay. The effect of miR211 and EZRIN overexpression on cell proliferation, migration/invasion, and apoptosis was detected. Results: The expression of miR211 was obviously lower in osteosarcoma tissues than paracancerous tissues. EZRIN was identified as the direct target of miR211, and up-regulation of miR211 increased the percentage of cell apoptosis, and suppressed cell proliferation as well as cell migration/invasion via directly regulating EZRIN. Conclusions: Our study indicated that miR211 has an important role in the development and progress of osteosarcoma, and it might become a novel target in the diagnosis and treatment of human osteosarcoma. |
| Keywords | miR211, EZRIN, Osteosarcoma, Cell viability, Cell apoptosis |
| Address and Contact Information |
1 Central laboratory, ZhongShan Hospital XiaMen University, Xiamen 361004, China. 2 Fujian Provincial Key Laboratory of Chronic Liver Disease and Hepatocellular Carcinoma (Xiamen University Affiliated ZhongShan Hospital), Xiamen 361004, China. 3 Department of Spine Surgery, ZhongShan Hospital XiaMen University, No. 201 Hubin South Road, Xiamen 361004, China. 4 Department of Rehabilitation, ZhongShan Hospital XiaMen University, Xiamen 361004, China. * Corresponding author: yaoqin20180101@sina.com †Yihua Pei and Qin Yao contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0172-y Volume 24 (2019) - 24:49 | |
| Title | SILENCING OF Synuclein-γ INHIBITS HUMAN CERVICAL CANCER THROUGH THE AKT SIGNALING PATHWAY |
| Authors | Chunnian Zhang*, Liqin Gu, Xiafang Li and Jianzhong Wang |
| Abstract | Background: Synuclein-γ has been demonstrated to be highly expressed in various human cancers including cervical cancer, and has been shown to play a critical role in tumor aggressiveness. We aimed to investigate the role of Synuclein-γ in human cervical cancer in vitro and in vivo. Method: Reverse transcription-quantitative polymerase chain reaction assay and Western blot assay were used to detect the mRNA and protein expression, respectively. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and colony formation assay were performed to measure the viabilities of cancer cells. Flow cytometry assay was used to detect the cell cycle and apoptosis. Moreover, an animal experiment was performed to evaluate the biological behavior of Synuclein-γ in vivo. Results: In the current study, we found that Synuclein-γ was obviously over-expressed in cervical cancer tissues compared to the adjacent non-cancer tissues. Cervical cancer cells transfected with Synuclein-γ siRNA demonstrated significant inhibition of cancer proliferation (P < 0.01), cell cycle arrest at G0/G1 phase, and cell apoptosis (P < 0.05). Moreover, down-regulation of Synuclein-γ significantly inhibited cervical cancer growth in vivo. In addition, protein levels of AKT, c-Myc and Cyclin D1 were much lower in the Synuclein-γ siRNA-treated groups than that in the control group. Conclusions: Synuclein-γ inhibition reduced cervical cancer tumor growth through the AKT pathway. This effect represented a therapeutic opportunity and provided a novel target for cervical cancer treatment. |
| Keywords | Synuclein-γ, Knockdown, Cervical cancer, Growth, AKT |
| Address and Contact Information | Department of Gynaecology, Ganzhou People’s Hospital of Jiangxi Province, No. 18, Meiguan Avenue, Ganzhou city 341000,
Jiangxi Province, China * Corresponding author: Chunnianzhang@163.com |
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| DOI: 10.1186/s11658-019-0175-8 Volume 24 (2019) - 24:50 | |
| Title | LONG NON-CODING RNA Malat1 ACTIVATED AUTOPHAGY, HENCE PROMOTING CELL PROLIFERATION AND INHIBITING APOPTOSIS BY SPONGING miR-101 IN COLORECTAL CANCER |
| Authors | Yaoran Si1, Zhaoguo Yang2, Quanxing Ge1, Lingbing Yu1, Meiying Yao1, Xinfang Sun1, Zheng Ren1 and Chunsheng Ding1* |
| Abstract | Background: Long non-coding RNA Malat1 has been widely identified as an oncogene which shows a significant relationship with tumorigenesis in colorectal cancer (CRC). Nonetheless, whether Malat1 participates in the autophagy of colorectal cancer remains unclear. Materials and methods: First, the expression level of Malat1 in 96 pairs of colorectal cancer tissues and four cell lines was detected by qRT-PCR. Subsequently, the autophagy activity in colorectal cancer tissues and cell lines was detected by western blot. Furthermore, the CCK-8 assay and flow cytometry (FCM) were performed to detect the role of autophagy activated by Malat1 in colorectal cancer cell lines. Results: In this study, significantly increased Malat1 expression and autophagy activity were found in colorectal cancer tissues compared with the adjacent normal tissues. Also, the Malat1 level was positively correlated with the expression of LC3-II mRNA in vivo. Moreover, autophagy activation and cell proliferation were significantly facilitated by Malat1 in colorectal cancer cells, while apoptosis decreased. Above all, the inhibition of autophagy by 3-MA not only relieved the Malat1-induced cell proliferation but also promoted the Malat1-induced cell apoptosis. In addition, Malat1 was found to act as an endogenous sponge by directly binding to miR-101 to reduce miR-101. Furthermore, the suppressive effects of miR-101 on the autophagy, proliferation, and apoptosis of CRC were abolished by Malat1. Conclusion: Long non-coding RNA Malat1 activated autophagy and promoted cell proliferation, yet inhibited apoptosis by sponging miR-101 in colorectal cancer cells. |
| Keywords | Colorectal cancer, Long noncoding RNA Malat1, Autophagy, Proliferation, Apoptosis, miR-101 |
| Address and Contact Information |
1 Department of Gastroenterology, Huaihe Hospital, Henan University, Kaifeng 475000, Henan, China 2 Department of General Surgery, Kaifeng Central Hospital, Kaifeng, Henan, China. * Corresponding author: ding_chunsheng@yeah.net |
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| DOI: 10.1186/s11658-019-0177-6 Volume 24 (2019) - 24:51 | |
| Title | MiR-135-5p PROMOTES OSTEOBLAST DIFFERENTIATION BY TARGETING HIF1AN IN MC3T3-E1 CELLS |
| Authors | Nuo Yin†, Longzhang Zhu†, Liang Ding, Junjie Yuan, Li Du, Mingmang Pan, Feng Xue* and Haijun Xiao* |
| Abstract | Background: MicroRNAs (miRNAs or miRs) serve crucial roles in the progression of osteoporosis. This study investigated the role and specific molecular mechanism of miR-135-5p in regulating osteoblast differentiation and calcification. Methods: Bone morphogenetic protein 2 (BMP2) was employed to interfere with the differentiation of MC3T3-E1. Then, miR-135-5p mimic or miR-135-5p inhibitor was transfected into MC3T3-E1, and quantitative RT-PCR was used to measure the expression of miR-135-5p. The expressions of runt-related transcription factor 2 (Runx2), osterix (OSX), osteopontin (OPN), and osteocalcin (OCN) were determined using western blot. Alkaline phosphatase (ALP) activity was measured using an appropriate kit assay. Calcium nodule staining was evaluated with alizarin red staining. A luciferase reporter assay was used to verify the target of miR-135-5p. Hypoxia-inducible factor 1 α inhibitor (HIF1AN) overexpression was applied to investigate its own role in the mechanism and a miR-135-5p rescue experiment was also performed. Results: Overexpression of miR-135-5p promoted osteogenic differentiation and calcification, as shown by the increase in ALP activity, calcification and osteogenic marker levels, including Runx2, OSX, OPN and OCN. Knockdown of miR-135-5p yielded the opposite results. HIF1AN was confirmed as a direct target of miR-135-5p. HIF1AN overexpression inhibited osteogenic differentiation and calcification while miR-135-5p reversed these effects. Conclusions: These results indicate that miR-135-5p might have a therapeutic application related to its promotion of bone formation through the targeting of HIF1AN. |
| Keywords | miR-135, Osteoblast differentiation, Calcification, Hypoxia-inducible factor 1 α inhibitor |
| Address and Contact Information | Department of Orthopaedics, Shanghai Fengxian District Central Hospital, No. 6600, Nanfeng Highway, Shanghai 201499, China
* Corresponding author: xuefengmd@sina.com; haijunxiao123@163.com; haijunxiao123@163.com †Nuo Yin and Longzhang Zhu contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0179-4 Volume 24 (2019) - 24:52 | |
| Title | COSTUNOLIDE REDUCES GLYCOLYSIS-ASSOCIATED ACTIVATION OF HEPATIC STELLATE CELLS VIA INHIBITION OF HEXOKINASE-2 |
| Authors | Dujing Ban1,3, Shangbo Hua2, Wen Zhang3 Chao Shen3, Xuehua Miao3 and Wensheng Liu3* |
| Abstract | Background: Hepatic stellate cell (HSC) activation is a central event during hepatic fibrosis. Aerobic glycolysis is one of its metabolic hallmarks. Blocking glycolysis is a novel therapeutic option for liver fibrosis. This study investigated the effects of costunolide, a natural product demonstrated to have hepatoprotective effects, on HSC activation and glycolysis. Methods: Primary HSCs were isolated from rats and cultured through 5 to 6 passages. Cell viability, activation markers, and glycolytic metabolism were examined in primary HSCs using various cellular and molecular approaches. Results: At 30 μM, costunolide reduced the viability of HSCs and inhibited the expression of α-smooth muscle actin and collagen I, two key markers of HSC activation. It also decreased glucose uptake and consumption, and reduced the intracellular levels of lactate in HSCs. At 10 mM, the glycolysis inhibitor 2-DG had a similar impact to costunolide at 30 μM: it significantly downregulated the expression of HSC activation markers. The combination of the two compounds produced more remarkable effects. Furthermore, costunolide repressed the expression and activity of hexokinase 2 (HK2), a pivotal rate-limiting enzyme that regulates glycolysis. However, overexpression of HK2 via plasmid transfection significantly reversed the costunolide-mediated downregulation of activation markers in HSCs, indicating that suppression of HK2 was required for costunolide to inhibit glycolysis-associated HSC activation. Conclusions: Our results show that costunolide can suppress HSC activation, and this is associated with inhibition of HK2, which blocks aerobic glycolysis. This suggests that costunolide is an antifibrotic candidate with potential for further development. |
| Keywords | Hepatic fibrosis, Hepatic stellate cell, Costunolide, Glycolysis, Hexokinase 2 |
| Address and Contact Information |
1 Nanjing University of Chinese Medicine, Nanjing 210023, China. 2 The Kunshan Hospital Affiliated to Nanjing University of Chinese Medicine, Kunshan 215300, China. 3 The Nanjing Integrative Medicine Hospital Affiliated to Nanjing University of Chinese Medicine, Nanjing 210014, China * Corresponding author: 18952681818@163.com |
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| DOI: 10.1186/s11658-019-0178-5 Volume 24 (2019) - 24:53 | |
| Title | CURRENT PREVALENCE STATUS OF GASTRIC CANCER AND RECENT STUDIES ON THE ROLES OF CIRCULAR RNAS AND METHODS USED TO INVESTIGATE CIRCULAR RNAs |
| Authors | Fei Jiang1,2*† and Xiaobing Shen1,2† |
| Abstract | Gastric cancer is a malignant tumor with the fifth incidence and third mortality worldwide. There were 951,000 new cases and about 723,000 patients died of it in 2012. Undoubtedly, gastric cancer has been affecting people’s living standards, and is already a major public health problem in China with its population growth and ageing. Even though the detection methods and medical standards have improved, the five-year survival rate of people is still very low. While circular RNA (circRNA) is increasingly attracting attention from researchers, at the same time, its mystery has gradually been uncovered. Many studies have shown that circRNA can act as molecular sponge of miRNA to regulate gene expression and has an obviously different expression profile between cancerous and normal groups, which arouse people’s curiosity and provide new opportunities for early detection of gastric cancer to improve the quality of life of patients. This study reviews current prevalence of gastric cancer in the word and China, as well as the characteristics and functions of circRNA and common laboratory detection methods involving circRNA in gastric cancer. |
| Keywords | Gastric cancer, Prevalence situation, CircRNA, Functions, Laboratory detection methods |
| Address and Contact Information |
1 Key Laboratory of Environmental Medical Engineering and Education Ministry, Nanjing Public Health College, Southeast University, Nanjing 210000, China 2 Department of Preventive Medicine, Nanjing Public Health College, Southeast University, Nanjing 210000, China * Corresponding author: seven_fly@foxmail.com † Fei Jiang and Xiaobing Shen contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0174-9 Volume 24 (2019) - 24:54 | |
| Title | A NOVEL FRAMESHIFT MUTATION IN THE EDA GENE IN AN IRANIAN PATIENT AFFECTED BY X-LINKED HYPOHIDROTIC ECTODERMAL DYSPLASIA |
| Authors | Marzieh Rahbaran1, Maryam Hassani Doabsari1, Simindokht Salavitabar1, Neda Mokhberian2, Ziba Morovvati3 and Saeid Morovvati4* |
| Abstract | Purpose: Ectodermal dysplasias are characterized by developmental abnormalities in ectodermal structures. Hypohidrotic ectodermal dysplasias (HED) are the most common subtype. They are most commonly inherited via X-linked recessive routes. We report on a novel ectodysplasin-A (EDA) mutation that is expected to be involved in pathogenesis of HED. Methods: Hypohidrotic ectodermal dysplasia genes, including EDA, EDAR and EDARADD, were analyzed using next-generation sequencing (NGS). The detected mutation on the EDA gene was confirmed in the patient and his mother using Sanger sequencing. Results: The patient presented with adontia, absence of gum development, hyperthermia and hypohidrosis. Our genetic analysis of the patient revealed a novel frameshift hemizygous mutation (c.898_924 + 8del35ins4CTTA) on the (EDA) gene. The patient’s mother showed a mild HED phenotype. Direct sequencing of the (EDA) gene in the region where her son had the mutation showed the same mutation in a heterozygous state. Conclusion: We identified a novel frameshift mutation in the (EDA) gene in an Iranian patient affected by X-linked HED. The difference between our patient’s symptoms and those recorded for some previous subjects may be due to the differences in the mutations involved. |
| Keywords | (EDA), Gene, Hypohidrotic, Ectodermal, Dysplasia |
| Address and Contact Information |
1 Islamic Azad Tehran Medical Sciences University, Tehran, Iran. 2 Department of biotechnology, School of Advanced Technology in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. 3 Department of Medical Genetics, Faculty of Advanced Medical Technologies, Golestan University of Medical Sciences, Gorgan, Iran. 4 Human Genetics Research Center, Baqiyatallah University of Medical Sciences, Mollasadra St, Tehran, Iran * Corresponding author: morovvati@bmsu.ac.ir; morovvati@hotmail.com |
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| DOI: 10.1186/s11658-019-0180-y Volume 24 (2019) - 24:55 | |
| Title | SOX2 PROMOTES HYPOXIA-INDUCED BREAST CANCER CELL MIGRATION BY INDUCING NEDD9 EXPRESSION AND SUBSEQUENT ACTIVATION OF Rac1/HIF-1α SIGNALING |
| Authors | Yueyuan Wang1, Maria Bibi1, Pengxiang Min1, Wenjie Deng1, Yujie Zhang1,2 and Jun Du1,2* |
| Abstract | Background: Hypoxia, a major condition associated with the tumor microenvironment, stimulates the migration of cancer cells. SOX2 is a powerful transcription factor that shows higher expression in several cancers, however, its role in hypoxia-induced breast cancer cell migration remains largely elusive. Methods: The human breast cancer cell lines MDA-MB-231 and MDA-MB-468 were cultured under hypoxic conditions. The cell migration rate was determined using the wound-healing and transwell assays. The protein levels of SOX2, NEDD9 and HIF-1α were evaluated via western blotting analysis. The NEDD9 mRNA levels were evaluated using qPCR. The activation of Rac1 was detected with the pulldown assay. The binding of SOX2 to the NEDD9 promoter was checked using the luciferase reporter assay. We also transfected breast cancer cells with specific siRNA for SOX2, NEDD9 or the Rac1 inactive mutant (T17 N) to investigate the role of SOX2, NEDD9 and Rac1 in the response to hypoxia. Results: Hypoxia markedly increased SOX2 protein levels in a time-dependent manner. SiRNA-mediated disruption of SOX2 inhibited cell migration under hypoxic conditions. Hypoxia also significantly augmented the NEDD9 mRNA and protein levels. Interestingly, SOX2 is a positive transcriptional regulator of NEDD9. Knockdown of SOX2 inhibited hypoxia-induced NEDD9 mRNA and protein expressions. Furthermore, hypoxia-induced upregulation of Rac1 activity and HIF-1α expression was attenuated by SOX2 or NEDD9 silencing, and Rac1-T17 N abolished HIF-1α expression as well as cell migration in cells subjected to hypoxia. Conclusions: Our results highlight the essential role of SOX2 in breast cancer cell motility. The upregulation of SOX2 under hypoxic conditions may facilitate NEDD9 transcription and expression, and subsequent activation of Rac1 and HIF-1α expression. This could accelerate breast cancer cell migration. |
| Keywords | Hypoxia, Migration, Breast cancer cells, SOX2, NEDD9 |
| Address and Contact Information |
1 Department of Physiology, Nanjing Medical University, 101 Longmian Avenue, Jiangning District, Nanjing 211166, Jiangsu, China 2 Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center for Personalized Cancer Medicine, Nanjing Medical University, 101 Longmian Avenue, Jiangning District, Nanjing 211166, Jiangsu, China * Corresponding author: dujun@njmu.edu.cn |
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| DOI: 10.1186/s11658-019-0176-7 Volume 24 (2019) - 24:56 | |
| Title | CORRECTION TO: MESENCHYMAL STEM CELLS DECREASE BLOOD–BRAIN BARRIER PERMEABILITY IN RATS WITH SEVERE ACUTE PANCREATITIS |
| Authors | Ronggui Lin1†, Ming Li2†, Meiqin Luo3, Tianhong Teng1, Yu Pan1 and Heguang Huang1* |
| Abstract | Correction to: Cell Mol Biol Lett (2019) 24:43 https://doi.org/10.1186/s11658-019-0167-8 Following publication of the original article, the author informed us that Fig. 5 was incorrect. |
| Keywords | |
| Address and Contact Information |
1 Department of General surgery, Fujian Medical University Union Hospital, 29 Xinquan Road, Fuzhou, Fujian 350001, People’s Republic of China 2 Department of Histology and Embryology, Hunan University of Medicine, Huaihua, Hunan, China. 3 Department of Orthopedics, Fujian Medical University Union Hospital, Fuzhou, Fujian, China. * Corresponding author: Heguanghuang2@163.com †Ronggui Lin and Ming Li contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0181-x Volume 24 (2019) - 24:57 | |
| Title | GRP78 REGULATES MILK BIOSYNTHESIS AND THE PROLIFERATION OF BOVINEMAMMARYEPITHELIAL CELLS THROUGH THE mTOR SIGNALING PATHWAY |
| Authors | Ying Liu1, Xuemei Wang2, Zhen Zhen1, Yanbo Yu1, Youwen Qiu1* and Wensheng Xiang1 |
| Abstract | Background: Glucose-regulated protein 78 (GRP78) is a member of the HSP70 protein family and a key endoplasmic reticulum chaperone. It has been revealed to play important roles both in the maturation, folding and transport of proteins and in cellproliferation. However, its involvement in milk biosynthesis or the proliferation of bovine primary mammary epithelial cells (BMECs) has yet to be established. Methods: The expressions of GRP78 in BMECs stimulated with methionine, leucine, estrogen and prolactin were determined using western blotting and immunofluorescence assays. To explore the function of GRP78 in BMECs, the protein was overexpressed or knocked down, respectively using an overexpression vector or an siRNA mixture transfected into cells cultured in vitro. Flow cytometry was used to analyze cell proliferation and cell activity. The contents of lactose and triglyceride (TG) secreted from the treated BMECs were measured using lactose and TG assay kits, respectively. Western blotting analysis was used to measure the β-casein content and the protein levels of the signaling molecules known to be involved in milk biosynthesis and cell proliferation. Results: GRP78overexpression significantly stimulated milk protein and milk fat synthesis, enhanced cell proliferation, positively regulated the phosphorylation of mammalian target of rapamycin (mTOR), and increased the amount of protein of cyclinD1andsterol regulatory element-binding protein 1c (SREBP-1c). GRP78 knockdown after siRNA transfection had the opposite effects. We further found that GRP78 was located in the cytoplasm of BMECs, and that stimulating methionine, leucine, estrogen and prolactin expression led to a significant increase in the protein expression of GRP78 in BMECs. Conclusions: These data reveal that GRP78 is an important regulator of milk biosynthesis and the proliferation of BMECs through the mTOR signaling pathway. |
| Keywords | Cyclin D1, GRP78, Bovine mammary epithelial cells, mTOR, SREBP-1c |
| Address and Contact Information |
1 The Key Laboratory of Dairy Science of Education Ministry, Heilongjiang Province, China 2 HaiNanUniversity, Hainan Province, China. * Corresponding author: yw12_630@126.com |
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| DOI: 10.1186/s11658-019-0182-9 Volume 24 (2019) - 24:58 | |
| Title | PAK4, A TARGET OF miR-9-5p, PROMOTES CELL PROLIFERATION AND INHIBITS APOPTOSIS IN COLORECTAL CANCER |
| Authors | Meihua Wang*, Qianqian Gao, Yufang Chen, Ziyan Li, Lingping Yue and Yun Cao |
| Abstract | Background: Colorectal cancer (CRC) is a leading cause of cancer-related death worldwide. P21-activated kinase 4 (PAK4) and miR-9-5p have emerged as attractive therapeutic targets in several tumor types, but in CRC, the regulation of their biological function and their target association remain unclear. Methods: The expression of PAK4 in CRC tissues was determined using quantitative real-time PCR and immunohistochemistry analyses. The targeted regulation between miR-9-5p and PAK4 was predicted and confirmed with bioinformatics analysis and the dual-luciferase reporter assay. Functional experiments, including the MTT assay and flow cytometry, were performed to investigate the impact of PAK4 knockdown and miR-9-5p overexpression on cell proliferation and apoptosis in CRC cells. Results: We found that the expression of PAK4 was upregulated in CRC tissues. PAK4 knockdown significantly suppressed cell proliferation and promoted apoptosis in cells of the CRC cell lines HCT116 and SW1116. We also found that miR-9-5p directly targeted the 3′-UTR of PAK4 mRNA and negatively regulated its expression. The degree of downregulation of miR-9-5p inversely correlated with PAK4 expression. Intriguingly, enforced expression of miR-9-5p suppressed cell proliferation and promoted apoptosis. This could be partially reversed by PAK4 overexpression. Conclusion: These results suggest that miR-9-5p targeting of PAK4 could have therapeutic potential for CRC treatment. |
| Keywords | Colorectal cancer, PAK4, Proliferation, Apoptosis, miR-9-5p |
| Address and Contact Information |
Department of Pathology, Changzhou Tumor Hospital Affiliated to Soochow University, 68 Honghe Road Changzhou, Changzhou 213032, Jiangsu, China * Corresponding author: mei_huaW18@126.com |
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| DOI: 10.1186/s11658-019-0183-8 Volume 24 (2019) - 24:59 | |
| Title | MONOAMINE OXIDASE-A ACTIVITY IS REQUIRED FOR CLONAL TUMORSPHERE FORMATION BY HUMAN BREAST TUMOR CELLS |
| Authors | William D. Gwynne, Mirza S. Shakeel, Jianhan Wu, Robin M. Hallett, Adele Girgis-Gabardo, Anna Dvorkin-Gheva and John A. Hassell* |
| Abstract | Background: Breast tumor growth and recurrence are driven by an infrequent population of breast tumor-initiating cells (BTIC). We and others have reported that the frequency of BTIC is orders of magnitude higher when breast tumor cells are propagated in vitro as clonal spheres, termed tumorspheres, by comparison to adherent cells. We exploited the latter to screen > 35,000 small molecules to identify agents capable of targeting BTIC. We unexpectedly discovered that selective antagonists of serotonin signaling were among the hit compounds. To better understand the relationship between serotonin and BTIC we expanded our analysis to include monoamine oxidase-A (MAO-A), an enzyme that metabolizes serotonin. Methods: We used the Nanostring technology and Western blotting to determine whether MAO-A is expressed in human breast tumor cell lines cultured as tumorspheres by comparison to those grown as adherent cells. We then determined whether MAO-A activity is required for tumorsphere formation, a surrogate in vitro assay for BTIC, by assessing whether selective MAO-A inhibitors affect the frequency of tumorsphere-forming cells. To learn whether MAO-A expression in breast tumor cells is associated with other reported properties of BTIC such as anticancer drug resistance or breast tumor recurrence, we performed differential gene expression analyses using publicly available transcriptomic datasets. Results: Tumorspheres derived from human breast tumor cell lines representative of every breast cancer clinical subtype displayed increased expression of MAO-A transcripts and protein by comparison to adherent cells. Surprisingly, inhibition of MAO-A activity with selective inhibitors reduced the frequency of tumorsphere-forming cells. We also found that increased MAO-A expression is a common feature of human breast tumor cell lines that have acquired anticancer drug resistance and is associated with poor recurrence-free survival (RFS) in patients that experienced high-grade, ER-negative (ER−) breast tumors. Conclusions: Our data suggests that MAO-A activity is required for tumorsphere formation and that its expression in breast tumor cells is associated with BTIC-related properties. The discovery that a selective MAO-A inhibitor targets tumorsphere-forming cells with potencies in the nanomolar range provides the first evidence of this agent’s anticancer property. These data warrant further investigation of the link between MAO-A and BTIC. |
| Keywords | Breast tumor-initiating cells, Monoamine oxidase-A, Tumorspheres |
| Address and Contact Information |
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada * Corresponding author: hassell@mcmaster.ca |
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| DOI: 10.1186/s11658-019-0188-3 Volume 24 (2019) - 24:60 | |
| Title | MiR-7-5p SUPPRESSES TUMOR METASTASIS OF NON-SMALL CELL LUNG CANCER BY TARGETING NOVA2 |
| Authors | Haiping Xiao |
| Abstract | Background: Non-small cell lung cancer (NSCLC) is the leading cause of cancer mortality worldwide. Distant metastasis is thought to be one of the most important factors responsible for the failure of NSCLC therapy. MicroRNA-7-5p (miR-7-5p) has been demonstrated to be a tumor suppressor in breast cancer, hepatocarcinoma, prostate cancer and glioblastoma multiforme (GBM). However, its role in NSCLC is still not fully understood. This study evaluated the role of miR-7-5p in the progression of NSCLC and explored the underlying mechanism. Materials & methods: The quantitative real-time PCR (qPCR), MTT, migration and invasion assays were used to evaluate the effects of miR-7-5p on the proliferation, migration and invasion of A549 and SPCA-1 cells. A tumor xenograft model was created to determine the effects of miR-7-5p on metastasis in vivo. The dual-luciferase reporter gene, neuro-oncological ventral antigen 2 (NOVA2) overexpression and western blotting assays were performed to explore the underlying mechanism. Results: MiR-7-5p is downregulated in NSCLC tissues and lung cancer cell lines. It suppresses proliferation, migration, invasion and EMT marker expression in vitro and in vivo. Further study showed that miR-7-5p suppresses tumor metastasis of NSCLC by targeting NOVA2. Overexpression of NOVA2 attenuates the miR-7-5p-mediated inhibitory effect on lung cancer cells. Conclusion: MiR-7-5p suppresses NSCLC metastasis. Targeting miR-7-5p may contribute to the success of NSCLC therapy. |
| Keywords | Non-small cell lung cancer (NSCLC), microRNA-7-5p (miR-7-5p), Suppress, Metastasis, Neuro-oncological ventral antigen 2 (NOVA2) |
| Address and Contact Information |
Thoracic Surgery Department, General Hospital of Southern Theater Command, Guangzhou 510010, PLA, China Corresponding author: XHP2005XHP@126.com |
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| DOI: 10.1186/s11658-019-0187-4 Volume 24 (2019) - 24:61 | |
| Title | DOWN-REGULATION OF miR-30b-5p PROTECTS CARDIOMYOCYTES AGAINST HYPOXIA-INDUCED INJURY BY TARGETING AVEN |
| Authors | Lanfang Zhang* and Xinwei Jia |
| Abstract | Background: Ischemia/hypoxia-induced cardiomyocyte apoptosis has been considered as a main cause of myocardial infarction. Here, we aimed to investigate the functional role of miR-30b-5p in hypoxic cardiomyocytes. Methods: AC16 human cardiomyocytes were cultured under hypoxia to simulate myocardial infarction. A qRT-PCR assay was performed to determine miR-30b-5p expression in hypoxic cardiomyocytes. Cell survival, injury and apoptosis were assessed by MTT, lactate dehydrogenase (LDH) release, and flow cytometry assays, respectively. The target gene of miR-30b-5p in hypoxic cardiomyocytes was validated by luciferase reporter assay and Western blotting. Results: MiR-30b-5p expression was found to be significantly upregulated in hypoxic AC16 cells. The in vitro experiments showed that downregulation of miR-30b-5p effectively alleviated hypoxia-induced cardiomyocyte injury. Furthermore, Aven is a potential target gene of miR-30b-5p and its downregulation could partially reverse the influence of miR-30b-5p knockdown on AC16 cells under hypoxia. Conclusions: Inhibition of miR-30b-5p could protect cardiomyocytes against hypoxia-induced injury by targeting Aven. |
| Keywords | Myocardial infarction, Hypoxia-induced injury, miR-30b-5p, Aven |
| Address and Contact Information |
Department of Cardiology, Affiliated Hospital of Hebei University, No. 212 Yuhua East Road, Baoding 071000, Hebei, People’s Republic of China * Corresponding author: lanfang_zhang12@126.com |
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| DOI: 10.1186/s11658-019-0186-5 Volume 24 (2019) - 24:62 | |
| Title | MicroRNA-146a PROTECTS AGAINST MYOCARDIAL ISCHAEMIA REPERFUSION INJURY BY TARGETING Med1 |
| Authors | Tiantian Zhang1†, Yiwen Ma2†, Lin Gao1, Chengyu Mao1, Huasu Zeng1, Xiaofei Wang1, Yapin Sun1, Jianmin Gu3, Yue Wang1, Kan Chen1, Zhihua Han1, Yuqi Fan1, Jun Gu1, Junfeng Zhang1* and Changqian Wang1* |
| Abstract | Background: Myocardial ischaemia reperfusion injury (MIRI) is a difficult problem in clinical practice, and it may involve various microRNAs. This study investigated the role that endogenous microRNA-146a plays in myocardial ischaemia reperfusion and explored the possible target genes. Methods: MIRI models were established in microRNA-146a deficient (KO) and wild type (WT) mice. MicroRNA-146a expression was evaluated in the myocardium of WT mice after reperfusion. The heart function, area of myocardium infarction and in situ apoptosis were compared between the KO and WT mice. Microarray was used to explore possible target genes of microRNA-146a, while qRT-PCR and dual luciferase reporter assays were used for verification. Western blotting was performed to detect the expression levels of the target gene and related signalling molecules. A rescue study was used for further testing. Results: MicroRNA-146a was upregulated 1 h after reperfusion. MicroRNA-146a deficiency decreased heart function and increased myocardial infarction and apoptosis. Microarray detected 19 apoptosis genes upregulated in the KO mice compared with the WT mice. qRT-PCR and dual luciferase verified that Med1 was one target gene of microRNA-146a. TRAP220, encoded by Med1 in the KO mice, was upregulated, accompanied by an amplified ratio of Bax/Bcl2 and increased cleaved caspase-3. Inhibition of microRNA-146a in H9C2 cells caused increased TRAP220 expression and more apoptosis under the stimulus of hypoxia and re-oxygenation, while knockdown of the increased TRAP220 expression led to decreased cell apoptosis. Conclusions: MicroRNA-146a exerts a protective effect against MIRI, which might be partially mediated by the target gene Med1 and related to the apoptosis signalling pathway. |
| Keywords | Myocardial ischaemia reperfusion injury, Apoptosis, microRNA-146a, Med1 |
| Address and Contact Information |
1 Department of Cardiology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China 2 Department of Anaesthesiology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China. 3 Department of Cardiovascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China. * Corresponding author: jfzhang_dr@163.com; wcqian@hotmail.com † Tiantian Zhang and Yiwen Ma contributed equally to this work. |
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| DOI: 10.1186/s11658-019-0185-6 Volume 24 (2019) - 24:63 | |
| Title | EXPRESSION PROFILES OF LONG NONCODING RNAs AND MESSENGER RNAs IN THE BORDER ZONE OF MYOCARDIAL INFARCTION IN RATS |
| Authors | Qingkun Meng, Zhijun Sun* , Hui Gu, Jiaying Luo, Jingjing Wang, Chuanhe Wang and Su Han |
| Abstract | Background: The participation of long noncoding RNAs (lncRNAs) in myocardial infarction has recently been noted. However, their underlying roles in the border zone of myocardial infarction remain unclear. This study uses microarrays to determine the profiles of lncRNAs and mRNAs in the border zone. Methods: Bioinformatics methods were employed to uncover their underlying roles. Highly dysregulated lncRNAs was further validated via PCR. Results: Four hundred seven lncRNAs and 752 mRNAs were upregulated, while 132 lncRNAs and 547 mRNAs were downregulated in the border zone of myocardial infarction. A circos graph was constructed to visualize the chromosomal distribution and classification of the dysregulated lncRNAs and mRNAs. The upregulated mRNAs in the border zone were most highly enriched in cytokine activity, binding, cytokine receptor binding and related processes, as ascertained through Go analysis. Pathway analysis of the upregulated mRNAs showed the most significant changes were in the TNF signaling pathway, cytokine–cytokine receptor interaction and chemokine signaling pathway and similar pathways and interactions. An lncRNA–mRNA co-expression network was established to probe into the underlying functions of the 10 most highly dysregulated lncRNAs based on their co-expressed mRNAs. In the co-expression network, we found 16 genes directly involved in myocardial infarction, including Alox5ap, Itgb2 and B4galt1. The lncRNAs AY212271, EF424788 and MRAK088538, among others, might be associated with myocardial infarction. BC166504 is probably a key lncRNA in the border zone of myocardial infarction. Conclusions: The results may have revealed some aberrantly expressed lncRNAs and mRNAs that contribute to the underlying pathophysiological mechanisms of myocardial infarction. |
| Keywords | Long noncoding RNAs, mRNAs, Myocardial infarction, Border zone, Area at risk, Co-expression network, Bioinformation |
| Address and Contact Information | Shengjing Hospital, China Medical University, Shenyang, China * Corresponding author: 565910412@qq.com |
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| DOI: 10.1186/s11658-019-0191-8 Volume 24 (2019) - 24:64 | |
| Title | CELLULAR HYPOXIA PROMOTES OSTEOGENIC DIFFERENTIATION OF MESENCHYMAL STEM CELLS AND BONE DEFECT HEALING VIA STAT3 SIGNALING |
| Authors | Xin Yu1,2,3†, Qilong Wan2†, Xiaoling Ye1, Yuet Cheng1, Janak L. Pathak4* and Zubing Li1,2* |
| Abstract | Background: Hypoxia in the vicinity of bone defects triggers the osteogenic differentiation of precursor cells and promotes healing. The activation of STAT3 signaling in mesenchymal stem cells (MSCs) has similarly been reported to mediate bone regeneration. However, the interaction between hypoxia and STAT3 signaling in the osteogenic differentiation of precursor cells during bone defect healing is still unknown. Methods: In this study, we assessed the impact of different durations of CoCl2-induced cellular hypoxia on the osteogenic differentiation of MSCs. Role of STAT3 signaling on hypoxia induced osteogenic differentiation was analyzed both in vitro and in vivo. The interaction between cellular hypoxia and STAT3 signaling in vivo was investigated in a mouse femoral bone defect model. Results: The peak osteogenic differentiation and expression of vascular endothelial growth factor (VEGF) occurred after 3 days of hypoxia. Inhibiting STAT3 reversed this effect. Hypoxia enhanced the expression of hypoxia-inducible factor 1-alpha (HIF-1α) and STAT3 phosphorylation in MSCs. Histology and μ-CT results showed that CoCl2 treatment enhanced bone defect healing. Inhibiting STAT3 reduced this effect. Immunohistochemistry results showed that CoCl2 treatment enhanced Hif-1α, ALP and pSTAT3 expression in cells present in the bone defect area and that inhibiting STAT3 reduced this effect. Conclusions: The in vitro study revealed that the duration of hypoxia is crucial for osteogenic differentiation of precursor cells. The results from both the in vitro and in vivo studies show the role of STAT3 signaling in hypoxia-induced osteogenic differentiation of precursor cells and bone defect healing. |
| Keywords | Cellular hypoxia, Mesenchymal stem cells, STAT3-signaling, Osteogenic differentiation, Bone defect healing |
| Address and Contact Information |
1 The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine, Ministry of Education, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079, China 2 Department of Oral and Maxillofacial Trauma and Plastic Surgery, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079, China. 3 Department of Stomatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. 4 Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Affiliated Stomatology Hospital of Guangzhou Medical University, Guangzhou 510140, China * Corresponding author: janakpathak@163.com; lizubing@whu.edu.cn Xin Yu and Qilong Wan are share first authorship. |
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| DOI: 10.1186/s11658-019-0194-5 Volume 24 (2019) - 24:65 | |
| Title | ESTROGEN STIMULATES SREBP2 EXPRESSION IN HEPATIC CELL LINES VIA AN ESTROGEN RESPONSE ELEMENT IN THE SREBP2 PROMOTER |
| Authors | Ye Meng* and Lu Zong |
| Abstract | Objective: Hypoestrogenism in women is strongly associated with menopause and it can lead to lipid disorder, which predisposes people to premature cardiovascular disease. However, the mechanism of lipid disorder remains unclear. Sterol regulatory element-binding protein 2 (SREBP2) is the key transcription factor regulating cholesterol metabolism. We hypothesize that estrogen regulates SREBP2 transcription through an estrogen response element (ERE) in the SREBP2 promoter region. Methods: Human hepatoblastoma cells (HepG2) were treated with dose-dependent concentrations of estradiol (E2) for 24 h. Then, SREBP2 expression was determined via real-time PCR and immunofluorescence. The expressions of the SREBP2 downstream target genes HMGCR and LDLR were determined via real-time PCR. Lipid secretion in the culture media of HepG2 cells was measured using ELISA. Through bioinformatics analysis, we identified high-scoring ERE-like sequences in the SREBP2 gene promoter. Chromatin immunoprecipitation analysis was used to confirm the ERE. DNA fragments of the putative or mutated ERE-like sequence were synthesized and ligated into pGL3-basic plasmid to construct the SREBP2 promoter luciferase reporter systems. SREBP2-Luciferase (SREBP2-Luc), SREBP2-Mutation (SREBP2-Mut) and the blank control were transfected into hepatic cell lines. Luciferase activities were measured using the dual-luciferase reporter assay system. Chromatin immunoprecipitation analysis and the luciferase reporter assay were repeated in human hepatoma cells (HuH-7). Results: We found that E2 dose-dependently increased the expression of SREBP2 in HepG2 cells and that the increased levels were blocked when treated with an estrogen receptor-alpha antagonist. Additionally, E2 increased both HMGCR and LDLR expression and lipid secretion in HepG2 cells. Notably, we identified a functional ERE in the SREBP2 gene promoter, to which E2 could specifically bind and induce transcription. Conclusions: An ERE was identified in the SREBP2 gene promoter. It mediates the regulation of SREBP2 expression by estrogen in hepatocytes. This study provides a mechanism to link cardiovascular disease with estrogen. |
| Keywords | Sterol regulatory element-binding protein 2, Estradiol, Transcription regulation, Lipid metabolism |
| Address and Contact Information | The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui 230001, People’s Republic of China * Corresponding author: mengye@ustc.edu.cn |
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| DOI: 10.1186/s11658-019-0184-7 Volume 24 (2019) - 24:66 | |
| Title | MicroRNA-582–3p negatively regulates cell proliferation and cell cycle progression in acute myeloid leukemia by targeting cyclin B2 |
| Authors | Haixia Li1,2, Xuefei Tian3*, Paoqiu Wang1, Mao Huang4, Ronghua Xu5 and Tian Nie5* |
| Abstract | Background: MicroRNAs (miRNAs) function as post-transcriptional gene expression regulators. Some miRNAs, including the recently discovered miR-582–3p, have been implicated in leukemogenesis. This study aimed to reveal the biological function of miR-582–3p in acute myeloid leukemia (AML), which is one of the most frequently diagnosed hematological malignancies. Methods: The expression of miR-582–3p was determined using quantitative real- time PCR in blood samples from leukemia patients and in cell lines. Cell proliferation and cell cycle distribution were analyzed using the CCK-8, colony formation and flow cytometry assays. The target gene of miR-582–3p was verified using a dual-luciferase reporter assay. The G2/M phase arrest-related molecule contents were measured using western blotting analysis. Results: We found miR-582–3p was significantly downregulated in the blood samples from leukemia patients and in the cell lines. MiR-582–3p overexpression significantly impaired cell proliferation and induced G2/M cell cycle arrest in THP-1 cells. Furthermore, cyclin B2 (CCNB2) was confirmed as a target gene of miR-582–3p and found to be negatively regulated by miR-582–3p overexpression. More importantly, CCNB2 knockdown showed suppressive effects on cell proliferation and cell cycle progression similar to those caused by miR-582–3p overexpression. The inhibitory effects of miR-582–3p overexpression on cell proliferation and cell cycle progression were abrogated by CCNB2 transfection. Conclusion: These findings indicate new functions and mechanisms for miR-582–3p in AML development. Further study could clarify if miR-582–3p and CCNB2 are potential therapeutic targets for the treatment of AML. |
| Keywords | Acute myeloid leukemia, miR-582–3p, Cell cycle, CCNB2 |
| Address and Contact Information |
1 Department of Integrated Chinese and Western Medicine, Hunan Children’s Hospital, Changsha 410007, China. 2 Hunan University of Chinese Medicine, Changsha 410208, China. 3 College of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha 410208, China. 4 Department of Pediatric Rehabilitation, Hebei Provincial Hospital of Traditional Chinese Medicine, Shijiazhuang 050000, Hebei, China. 5 Department of Hematology, The First Hospital of Hunan University of Chinese Medicine, 95 Shaoshan Middle Road, Changsha City 410007, Hunan Province, China. * Corresponding author: tian_Xf@outlook.com; nie_tian0607@126.com |
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| DOI: 10.1186/s11658-019-0189-2 Volume 24 (2019) - 24:67 | |
| Title | UPREGULATION OF miR-376c-3p ALLEVIATES OXYGEN–GLUCOSE DEPRIVATION-INDUCED CELL INJURY BY TARGETING ING5 |
| Authors | Heng Zhang, Jie Zhou, Mingxia Zhang, Yanjie Yi and Bing He* |
| Abstract | Background: The expression level of miR-376c-3p is significantly lower in infants with neonatal hypoxic-ischemic encephalopathy (HIE) than in healthy infants. However, the biological function of this microRNA remains largely elusive. Methods: We used PC-12 and SH-SY5Y cells to establish an oxygen–glucose deprivation (OGD) cell injury model to mimic HIE in vitro. The miR-376c-3p expression levels were measured using quantitative reverse transcription PCR. The CCK-8 assay and flow cytometry were utilized to evaluate OGD-induced cell injury. The association between miR-376c-3p and inhibitor of growth 5 (ING5) was validated using the luciferase reporter assay. Western blotting was conducted to determine the protein expression of CDK4, cyclin D1, Bcl-2 and Bax. Results: MiR-376c-3p was significantly downregulated in the OGD-induced cell injury model. Its overexpression elevated cell viability and impaired cell cycle G0/G1 phase arrest and apoptosis in PC-12 and SH-SY5Y cells after OGD. Downregulation of miR-376c-3p gave the opposite results. We further demonstrated that ING5 was a negatively regulated target gene of miR-376c-3p. Importantly, ING5 knockdown had a similar effect to miR-376c-3p-mediated protective effects against cell injury induced by OGD. Its overexpression abolished these protective effects. Conclusion: Our data suggest that miR-376c-3p downregulated ING5 to exert protective effects against OGD-induced cell injury in PC-12 and SH-SY5Y cells. This might represent a novel therapeutic approach for neonatal HIE treatment. |
| Keywords | miR-376c-3p, Oxygen–glucose deprivation, ING5, Cell cycle, Apoptosis |
| Address and Contact Information | Department of Pediatrics, Renmin Hospital of Wuhan University, Hubei Province 430060, China * Corresponding author: bing_heBEE@126.com |
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| DOI: 10.1186/s11658-019-0190-9 Volume 24 (2019) - 24:68 | |
| Title | MiR-214 PREVENTS THE PROGRESSION OF DIFFUSE LARGE B-CELL LYMPHOMA BY TARGETING PD-L1 |
| Authors | Jing-Ran Sun1, Xiao Zhang2 and Ya Zhang3* |
| Abstract | Objective: We explored the role and mechanism of miR-214 involvement in the progression of diffuse large B-cell lymphoma (DLBCL). Methods: The expression levels of miR-214 and PD-L1 in human DLBCL cell lines and in tissue samples from patients with DLBCL were determined using quantitative RT-PCR. The dual-luciferase reporter assay was employed to determine the correlation between the expressions of miR-214 and PD-L1. Cell viability, invasiveness and apoptosis were respectively examined in cells of the DLBCL line OCI-Ly3 using CCK-8, transwell and flow cytometry assays. The expression level of PD-L1 was determined via immunoblotting. Inflammatory cytokine secretion was determined via enzyme-linked immune sorbent assay (ELISA). Results: miR-214 was downregulated and PD-L1 was upregulated in DLBCL tissues and cell lines in comparison to normal adjacent tissues or normal B-cell. This indicates a negative correlation in the expression levels. Overexpression of miR-214 inhibited cell viability and invasion and induced apoptosis of OCI-Ly3 cells. Moreover, miR-214 was shown to target PD-L1 mRNA by binding to its 3′-untranslated region (UTR). Knockdown of PD-L1 attenuated the malignant phenotype of OCI-Ly3 cells. Overexpression of miR-214 inhibited tumor growth by targeting PD-L1 in vivo. Conclusion: By targeting PD-L1, miR-214 regulates the progression of DLBCL in vitro and in vivo. |
| Keywords | Diffuse large B-cell lymphoma, miR-214, PD-L1, Proliferation, Invasion, T cells |
| Address and Contact Information |
1 Liaocheng Central Blood Station, 75 Huashan Road, Liaocheng, Shandong 25200, People’s Republic of China. 2 Department of Clinical Laboratory, Liaocheng People’s Hospital, 67 Dongchang West Road, Liaocheng, Shandong 25200, People’s Republic of China. 3 Department of Gynecology and Obstetrics, Liaocheng People’s Hospital, 67 Dongchang West Road, Liaocheng, Shandong 25200, People’s Republic of China * Corresponding author: zhangya614@163.com |
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| DOI: 10.1186/s11658-019-0196-3 Volume 24 (2019) - 24:69 | |
| Title | EDITORIAL FOCUS: UNDERSTANDING OFF-TARGET EFFECTS AS THE KEY TO SUCCESSFUL RNAi THERAPY |
| Authors | Rafal Bartoszewski1* and Aleksander F. Sikorski2* |
| Abstract | With the first RNA interference (RNAi) drug (ONPATTRO (patisiran)) on the market, we witness the RNAi therapy field reaching a critical turning point, when further improvements in drug candidate design and delivery pipelines should enable fast delivery of novel life changing treatments to patients. Nevertheless, ignoring parallel development of RNAi dedicated in vitro pharmacological profiling aiming to identify undesirable off-target activity may slow down or halt progress in the RNAi field. Since academic research is currently fueling the RNAi development pipeline with new therapeutic options, the objective of this article is to briefly summarize the basics of RNAi therapy, as well as to discuss how to translate basic research into better understanding of related drug candidate safety profiles early in the process. |
| Keywords | ncRNAs, microRNAs, siRNAs, RNAi therapy, RNAi drug candidates, Off-target effects |
| Address and Contact Information |
1 Department of Biology and Pharmaceutical Botany, Medical University of Gdansk, Gdansk, Poland 2 Department of Cytobiochemistry, Faculty of Biotechnology, University of Wroclaw, Wroclaw, Poland * Corresponding author: rafalbar@gumed.edu.pl; aleksander.sikorski@uwr.edu.pl |
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| DOI: 10.1186/s11658-019-0192-7 Volume 24 (2019) - 24:70 | |
| Title | THE LONG NONCODING RNA LINC00483 PROMOTES LUNG ADENOCARCINOMA PROGRESSION BY SPONGING miR-204-3p |
| Authors | Shengzhuang Yang†, Tao Liu†, Yu Sun and Xiangsen Liang* |
| Abstract | Background: The expression of the long noncoding RNA LINC00483 is upregulated in lung adenocarcinoma (LUAD). However, its role in the progression of LUAD and the underlying mechanisms remain elusive. Methods: The expressions of LINC00483 and miR-204-3p were determined using quantitative real-time PCR. The correlation between the clinicopathological characteristics of LUAD patients and LINC00483 expression was analyzed using Pearson’s χ2 test. A549 and PC-9 cells were transfected with small interfering RNA (siRNA) that specially targeting LINC00483 to assess the impact of its knockdown. Cell proliferation was assessed using the Cell Counting Kit-8 and clone forming assays. Cell migration and cell invasion were evaluated using a transwell assay. The levels of Snail, E-cadherin, N-cadherin and ETS1 proteins were determined via western blotting. The interaction between LINC00483 and miR-204-3p was analyzed using dual-luciferase, fluorescence in situ hybridization and RNA immunoprecipitation. Results: LINC00483 was upregulated in LUAD tissues and cell lines. Higher LINC00483 levels closely correlated to shorter survival times, advanced TNM stage, larger tumor size and positive lymph node metastasis. Cell proliferation, migration and invasion were suppressed after LINC00483 knockdown. LINC00483 mainly localized in the cytoplasm, where it acted as a sponge of miR-204-3p. ETS1 was validated as a downstream target of miR-204-3p and is thus regulated by LINC00483. Conclusion: This study demonstrated that LINC00483 facilitates the proliferation, migration and invasion of LUAD cells by acting as a sponge for miR-204-3p, which in turn regulates ETS1. |
| Keywords | LINC00483, Lung adenocarcinoma, Cell proliferation, Cell migration/invasion, ETS1 |
| Address and Contact Information |
Department of Chest Cardiovascular Surgery, The Second Affiliated Hospital of Guangxi Medical University, 166 University East Road, Xixiangtang District, Nanning City
530007, Guangxi Province, China * Corresponding author: XiangsenLiangset@163.com † Shengzhuang Yang and Tao Liu contributed equally to the work. |
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| DOI: 10.1186/s11658-019-0195-4 Volume 24 (2019) - 24:71 | |
| Title | SOX30, A TARGET GENE OF miR-653-5p, REPRESSES THE PROLIFERATION AND INVASION OF PROSTATE CANCER CELLS THROUGH INHIBITION OF Wnt/β-catenin SIGNALING |
| Authors | Qiang Fu†, Zhenye Sun†, Fan Yang, Tianci Mao, Yanyao Gao* and He Wang* |
| Abstract | Background: Sex-determining region Y-box containing gene 30 (SOX30) is a newly identified tumor-associated gene in several types of cancer. However, whether SOX30 is involved in the development and progression of prostate cancer remains unknown. This study investigated the potential role of SOX30 in prostate cancer. Methods: Prostate cancer cell lines and a normal prostate epithelial cell line were used for the experiments. The expression of SOX30 was determined using quantitative real-time PCR and western blot analysis. The malignant cellular behaviors of prostate cancer were assessed using the Cell Counting Kit-8, colony formation and Matrigel invasion assays. The miRNA–mRNA interaction was validated using the dual-luciferase reporter assay. Results: SOX30 expression was lower in cells of prostate cancer lines than in cells of the normal prostate epithelial line. Its overexpression repressed the proliferation and invasion of prostate cancer cells. SOX30 was identified as a target gene of microRNA-653-5p (miR-653-5p), which is upregulated in prostate cancer tissues. MiR-653-5p overexpression decreased SOX30 expression, while its inhibition increased SOX30 expression in prostate cancer cells. MiR-653-5p inhibition also markedly restricted prostate cancer cell proliferation and invasion. SOX30 overexpression or miR-653-5p inhibition significantly reduced β-catenin expression and downregulated the activation of Wnt/β-catenin signaling. SOX30 knockdown significantly reversed the miR-653-5p inhibition-mediated inhibitory effect on the proliferation, invasion and Wnt/β-catenin signaling in prostate cancer cells. Conclusions: These results reveal a tumor suppressive function for SOX30 in prostate cancer and confirmed the gene as a target of miR-653-5p. SOX30 upregulation due to miR-653-5p inhibition restricted the proliferation and invasion of prostate cancer cells, and this was associated with Wnt/β-catenin signaling suppression. These findings highlight the importance of the miR-653-5p–SOX30–Wnt/β-catenin signaling axis in prostate cancer progression. |
| Keywords | SOX30, MiR-653-5p, Prostate cancer, Wnt/β-catenin |
| Address and Contact Information |
Department of Urology, Tangdu Hospital, The Fourth Military Medical University, 1 Xinsi Road, Xi’an 710038, Shaanxi, China * Corresponding author: yanyaogao@163. com; wang_hewh@163.com †Qiang Fu and Zhenye Sun share the first authorship. |
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| DOI: 10.1186/s11658-019-0198-1 Volume 24 (2019) - 24:72 | |
| Title | THE NOVEL CIRCULAR RNA circ-CAMK2A ENHANCES LUNG ADENOCARCINOMA METASTASIS BY REGULATING THE miR-615-5p/fibronectin 1 PATHWAY |
| Authors | Jiahui Du, Guangzhao Zhang, Hongli Qiu, Haifeng Yu and Wuying Yuan* |
| Abstract | Background: Circular RNA (circRNA) has recently been considered as a key regulator in carcinogenesis. In this study, we investigated the functional significance and regulatory role of circ-CAMK2A (hsa_circ_0128332) in lung adenocarcinoma (LUAD). Methods: GSE101586 was employed to screen differentially expressed circRNAs. = Relative expression levels of circ-CAMK2A, miR-615-5p, fibronectin 1 (FN1), MMP2, and MMP9 were tested by quantitative reverse transcription PCR (qRT-PCR) or western blotting. Functional experiments were performed by CCK-8, wound healing, and transwell assays. Luciferase reporter and biotin-labeled RNA pull-down assays were carried out to evaluate the interaction between circ-CAMK2A, miR-615-5p, and fibronectin 1. In addition, a lung metastasis model was constructed to determine the metastasis-promoting role of circ-CAMK2A in vivo. Results: Circ-CAMK2A overexpression was observed in LUAD and was closely associated with lymph node metastasis, distant metastasis, advanced clinical stage, and poor prognosis. Circ-CAMK2A silencing evidently inhibited LUAD cell migration and invasion, whereas circ-CAMK2A overexpression had an opposite effect. Importantly, overexpression of circ-CAMK2A also enhanced LUAD metastasis in vivo. Mechanistically, miR-615-5p was identified as a direct target of circ-CAMK2A. Circ-CAMK2A up-regulates the expression level of fibronectin 1 by sponging miR-615-5p, thereby increasing MMP2 and MMP9 expression to promote the metastasis of LUAD. Conclusion: Circ-CAMK2A plays a crucial role in the metastasis of LUAD, at least partially, by regulating the miR-615-5p/fibronectin 1 axis. |
| Keywords | Circular RNA, microRNA, Lung adenocarcinoma, Metastasis, Prognosis |
| Address and Contact Information |
Minimally invasive surgery, Henan Provincial Chest Hospital, No. 1 Weiwu Road, Jinshui District, Zhengzhou 450000, People’s Republic of China * Corresponding author: yuanwuying321@yeah.net |
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