Cellular & Molecular Biology Letters

International Scientific Journal
Established 1996

Volume 12 (2007) No. 3

DOI: 10.2478/s11658-007-0005-2 Volume 12 (2007) pp 317 - 330
Title	AN AUTORADIOGRAPHIC STUDY OF CELLULAR PROLIFERATON, DNA SYNTHESIS AND CELL CYCLE VARIABILITY IN THE RAT LIVER CAUSED BY PHENOBARBITAL-INDUCED OXIDATIVE STRESS: THE PROTECTIVE ROLE OF MELATONIN
Authors	Gamal H. El-Sokkary
Abstract	The protective effect of melatonin against phenobarbital-induced oxidative stress in the rat liver was measured based on lipid peroxidation levels (malondialedyde and 4-hydroxyalkenals). Cellular proliferation, DNA synthesis and cell cycle duration were quantitated by the incorporation of 3H-thymidine, detected by autoradiography, into newly synthesized DNA. Two experiments were carried out in this study, each on four equal-sized groups of male rats (control, melatonin [10 mg/kg], phenobabital [20 mg/kg] and phenobarbital plus melatonin). Experiment I was designed to study the proliferative activity and rate of DNA synthesis, and measure the levels of lipid peroxidation, while experiment II was for cell cycle time determination. Relative to the controls, the phenobarbital-treated rats showed a significant increase (P < 0.01) in the lipid peroxidation levels (30.7%), labelling index (69.4%) and rate of DNA synthesis (37.8%), and a decrease in the cell cycle time. Administering melatonin to the phenobarbital-treated rats significantly reduced (P < 0.01) the lipid peroxidation levels (23.5%), labelling index (38.2%) and rate of DNA synthesis (29.0%), and increased the cell cycle time. These results seem to indicate that the stimulatory effect of phenobarbital on the oxidized lipids, proliferative activity, kinetics of DNA synthesis and cell cycle time alteration in the liver may be one of the mechanisms by which the non-genotoxic mitogen induces its carcinogenic action. Furthermore, melatonin displayed powerful protection against the toxic effect of phenobarbital.
Keywords	Phenobarbital, Melatonin, Lipid peroxidation, Cell proliferation, DNA synthesis, Cell cycle
Adress and Contact Informations	Department of Zoology, Faculty of Science, Assiut University, Assiut, 71516, Egypt * Author for correspondence; e-mail: elsokkary2000@yahoo.com, tel: +20-88-2401724, fax: +20-88-2342708

DOI: 10.2478/s11658-007-0006-1 Volume 12 (2007) pp 331 - 347
Title	POST-TRANSCRIPTIONAL MODIFICATIONS OF VEGF-A mRNA IN NON-ISCHEMIC DILATED CARDIOMYOPATHY
Authors	Jacek Kowalczyk1*, Dorota Domal-Kwiatkowska², Urszula Mazurek³, Michał Zembala¹, Bogdan Michalski⁴ and Marian Zembala⁵
Abstract	Vascular endothelial growth factor (VEGF-A) is one of the most important proangiogenic factors. It has many isoforms encoded by one gene. The occurrence of these isoforms is associated with the process of alternative splicing of mRNA. Some of the splice forms are perceived as tissue specific. The aim of this study was to determine the alternative splicing of VEGF-A mRNA in dilated cardiomyopathy, especially at the level of particular myocardial layers. The assessment of post-transcriptional modifications of VEGF-A mRNA was made on specimens taken from the explanted hearts of patients undergoing cardiac transplantation. Molecular and histopathological studies were perfomed on particular layers of the myocardial muscle (endocardium, myocardium, epicardium). A molecular analysis of cardiac samples was performed by quantitative analysis of the mRNA of the studied VEGF-A isoforms (VEGF121, -145, -165, -183, -189, and -206) using QRT- PCR with an ABI-PRISM 7700-TaqMan sequence detector. 72 cardiac specimens taken from the explanted hearts were analyzed. Each of the studied VEGF-A splice forms was present in the evaluated hearts, but the types of alternative splicing of mRNA were different in particular layers. Quantitative analysis revealed different amounts of the studied isoforms. Generally, significantly increased expression of the VEGF-A isoforms was observed in samples taken from hearts with post-inflammatory etiology of cardiomyopathy. Our conclusions are: 1. All the studied VEGF-A isoforms were found in the human hearts, including those thusfar considered characteristic for other tissues. 2. Significant differences were observed in the expression of the VEGF-A splice forms with respect to the myocardial layers and the location of the cardiac biopsy. 3. Repetitive and comparable results for samples with post-inflammatory etiology were obtained, and they revealed considerably higher amounts of VEGF-A isoforms compared to specimens with idiopathic etiology.
Keywords	Vascular endothelial growth factor (VEGF), Alternative splicing, Angiogenesis, Dilated cardiomyopathy, Transcriptional activity
Adress and Contact Informations	¹1^st Department of Cardiology, Silesian Center for Heart Diseases, Zabrze, Medical University of Silesia, Katowice, Poland, ²Department of Biochemistry, Sosnowiec, Medical University of Silesia, Katowice, Poland, ³Department of Molecular Biology and Genetics, Sosnowiec, Medical University of Silesia, Katowice, Poland, ⁴Department of Gynaecology and Obstetrics, Tychy, Medical University of Silesia, Katowice, Poland, ⁵Department of Cardiac Surgery and Transplantology, Silesian Center for Heart Diseases, Zabrze, Medical University of Silesia, Katowice, Poland * Author for correspondence; e-mail: jacekmed@poczta.onet.pl, tel: (+48) 32 271-34-14, fax: (+48) 32 271-76-92

DOI: 10.2478/s11658-007-0007-0 Volume 12 (2007) pp 348 - 361
Title	GENISTEIN INHIBITS THE CONTACT-STIMULATED MIGRATION OF PROSTATE CANCER CELLS
Authors	Katarzyna Miękus and Zbigniew Madeja*
Abstract	The results of several epidemiological studies have suggested that a soybean-based diet is associated with a lower risk of prostate cancer. We investigated the effect of the soy isoflavone genistein on the proliferation and contact-stimulated migration of rat prostatic carcinoma MAT-LyLu and AT-2 cell lines. Genistein almost completely inhibited the growth of both MAT-LyLu and AT-2 cells in the concentration range from 25 to 100 µM, but the addition of 1 µM genistein to the medium significantly stimulated the proliferation of both cell lines. Additionally, at concentrations above 25 µM, genistein showed a potent cytotoxic effect. However, the central finding of this study is that at physiologically relevant concentrations (1 µM and 10 µM), genistein inhibits the motility of prostate cancer cells stimulated by homo- and heterotypic contacts. These results show that at physiological concentrations, genistein exerts an inhibitory effect on the migration of prostate cancer cells and suggest that it may be one of the factors responsible for the anti-metastatic activity of plant isoflavonoids
Keywords	Cell movement, Metastasis, Contact-stimulation, Prostate cancer, Genistein
Adress and Contact Informations	< Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30-387 Kraków, Poland * Author for correspondence; e-mail address: zibi@mol.uj.edu.pl

DOI: 10.2478/s11658-007-0008-z Volume 12 (2007) pp 362 - 369
Title	IMPROVED FUSION PROTEIN EXPRESSION OF EGFP VIA THE MUTATION OF BOTH KOZAK AND THE INITIAL ATG CODON
Authors	Chao Dai^#, Zhijian Cao^#, Yingliang Wu, Hong Yi, Dahe Jiang and Wenxin Li*
Abstract	Since its discovery, green fluorescence protein (GFP) has been used as a reporter in a broad range of applications, including the determination of gene expresion in diverse organisms, and subcellular protein localization. pEGFP-N1 is a eukayotic expression vector encoding EGFP, the MCS of which locates at the N terminus of EGFP. In this study, the cDNA sequence of scorpion toxin BmKK2 was inserted into the XhoI-HindIII cut of pEGFP-N1 to construct a toxin-EGFP fusion gene (named pEGFP-BmKK2). Fluorescence imaging revealed that HEK 293T cells that were transfected by pEGFP-BmKK2 emitted green fluorescence. Transcription of pEGFP-BmKK2 was confirmed by RT-PCR. However, western blotting analysis showed that the transfected HEK 293T cells expressed mostly EGFP, but little toxin-EGFP fusion protein, implying that pEGFP-N1 cannot be used as a fusion expression vector for subcellular protein localization for the BmKK2 gene. Consequently, two modified recombinant vectors (pEGFP-BmKK2-M1 and pEGFP-BmKK2-M2) were constructed based on pEGFP-BmKK2. This greatly improved the expression of toxin-EGFP fusion protein from pEGFP-BmKK2-M2.
Keywords	EGFP, Fusion protein expression, Subcellular protein localization, Scorpion toxin
Adress and Contact Informations	State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, P R China ^# These authors contributed equally to this article * Author for correspondence; e-mail: liwxlab@whu.edu.cn, tel: 86-27-68752831, fax: 86-27-68752146

DOI: 10.2478/s11658-007-0009-y Volume 12 (2007) pp 370 - 377
Title	HIGH LEVEL EXPRESSION OF STAG1/PMEPA1 IN AN ANDROGEN- INDEPENDENT PROSTATE CANCER PC3 SUBCLONE
Authors	Yoshifumi S. Hirokawa¹*, Akimitsu Takagi¹, Katsunori Uchida¹, Yuji Kozuka¹, Misao Yoneda¹, Masatoshi Watanabe² and Taizo Shiraishi¹
Abstract	In this paper, we describe the isolation and characterization of two PC3 subclones. One subclone, mr, showed an epithelial phenotype, the other, M1, showed a sarcomatous morphology. Transplanted into nude mice, mr developed tumors at a dramatically faster rate than M1. Comparing the two subclones, differentially expressed genes were identified, including E-cadherin, IL-8 and STAG1/PMEPA1. These genes were expressed at higher levels in mr than in M1.
Keywords	STAG1/PMEPA1, IL-8, E-cadherin, Prostate cancer cell line
Adress and Contact Informations	¹ Department of Pathological Oncology, Institute of Molecular and Experimental Medicine, Faculty of Medicine, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu, Mie 514-8507, Japan, ²Laboratory for Medical Engineering, Division of Materials Science and Chemical Engineering, Graduate School of Engineering, Yokohama National University, Yokohama, Japan. *Author for correspondence; fax: +81-59-231-5210; e-mail: ultray2k@clin.medic.mie-u.ac.jp

DOI: 10.2478/s11658-007-0010-5 Volume 12 (2007) pp 378 - 395
Title	ULTRACENTRIFUGATION STUDIES OF THE LOCATION OF THE SITE INVOLVED IN THE INTERACTION OF PIG HEART LACTATE DEHYDROGENASE WITH ACIDIC PHOSPHOLIPIDS AT LOW pH. A COMPARISON WITH THE MUSCLE FORM OF THE ENZYME
Authors	Grzegorz Terlecki*, Elżbieta Czapińska and Katarzyna Hotowy
Abstract	Lactate dehydrogenase (LDH) from the pig heart interacts with liposomes made of acidic phospholipids most effectively at low pH, close to the isoelectric point of the protein (pH = 5.5). This binding is not observed at neutral pH or high ionic strength. LDH-liposome complex formation requires an absence of nicotinamide adenine dinucleotides and adenine nucleotides in the interaction environment. Their presence limits the interaction of LDH with liposomes in a concentration-dependent manner. This phenomenon is not observed for pig skeletal muscle LDH. The heart LDH-liposome complexes formed in the absence of nicotinamide adenine dinucleotides and adenine nucleotides are stable after the addition of these substances even in millimolar concentrations. The LDH substrates and studied nucleotides that inhibit the interaction of pig heart LDH with acidic liposomes can be ordered according to their effectiveness as follows: NADH > NAD > ATP = ADP > AMP > pyruvate. The phosphorylated form of NAD (NADP), nonadenine nucleotides (GTP, CTP, UTP) and lactate are ineffective. Chemically cross-linked pig heart LDH, with a tetrameric structure stable at low pH, behaves analogously to the unmodified enzyme, which excludes the participation of the interfacing parts of subunits in the interaction with acidic phospholipids. The presented results indicate that in lowered pH conditions, the NADH-cofactor binding site of pig heart LDH is strongly involved in the interaction of the enzyme with acidic phospholipids. The contribution of the ATP/ADP binding site to this process can also be considered. In the case of pig skeletal muscle LDH, neither the cofactor binding site nor the subunit interfacing areas seem to be involved in the interaction.
Keywords	Lipid-protein interaction, Lactate dehydrogenase isoenzymes, Acidic phospholipids, Cardiolipin, Phosphatidylserine.
Adress and Contact Informations	Department of Medical Biochemistry, Wrocław Medical University, Wrocław, Poland * Author for correspondence; e-mail: terlecki@bioch.am.wroc.pl, tel: +48 (71) 784-13-81

DOI: 10.2478/s11658-007-0011-4 Volume 12 (2007) pp 396 - 406
Title	ESTROGEN REGULATION AND ION DEPENDENCE OF TAURINE UPTAKE BY MCF-7 HUMAN BREAST CANCER CELLS
Authors	David B. Shennan* and Jean Thomson
Abstract	It has been reported that estrogen receptor-positive MCF-7 cells express TauT, a Na⁺-dependent taurine transporter. However, there is a paucity of information relating to the characteristics of taurine transport in this human breast cancer cell line. Therefore, we have examined the characteristics and regulation of taurine uptake by MCF-7 cells. Taurine uptake by MCF-7 cells showed an absolute dependence upon extracellular Na+. Although taurine uptake was reduced in Cl^- free medium a significant portion of taurine uptake persisted in the presence of NO3 ^- . Taurine uptake by MCF-7 cells was inhibited by extracellular ß-alanine but not by L-alanine or L-leucine. 17ß-estadiol increased taurine uptake by MCF-7 cells: the Vmax of influx was increased without affecting the Km. The effect of 17ß-estradiol on taurine uptake by MCF-7 cells was dependent upon the presence of extracellular Na+. In contrast, 17ß-estradiol had no significant effect on the kinetic parameters of taurine uptake by estrogen receptor- negative MDA-MB-231 cells. It appears that estrogen regulates taurine uptake by MCF-7 cells via TauT. In addition, Na⁺-dependent taurine uptake may not be strictly dependent upon extracellular Cl^-.
Keywords	Breast cancer, Taurine uptake, Estrogen
Adress and Contact Informations	Strathclyde Institute of Pharmacy and Biomedical Sciences, Royal College, University of Strathclyde, 204 George Street, Glasgow, UK G1 1XW * Author for correspondence: e-mail: david.shennan@strath.ac.uk

DOI: 10.2478/s11658-007-0012-3 Volume 12 (2007) pp 407 - 421
Title	THE CONSTRUCTION OF THE EUKARYOTIC EXPRESSION PLASMID pcDNA3.1/AZURIN AND THE INCREASED APOPTOSIS OF U2OS CELLS TRANSFECTED WITH IT
Authors	Zhaoming Ye¹, Huiqin Peng², Yongming Fang³, Jie Feng¹ and Di-Sheng Yang¹*
Abstract	In our previous study, we demonstrated that azurin could selectively trigger apoptosis in human osteosarcoma cell line U2OS cells. However, the rate of apoptosis (35.8 ± 3.2%) is not very high, and azurin is too expensive to obtain readily. To solve these problems, we constructed a eukaryotic expression plasmid containing the azurin gene with an influenza virus haemagglutinin 9 peptide HA epitope tag, and transfected the recombinant plasmid pcDNA3.1(+)/azurin into U2OS cells. RT-PCR and Western blot analysis validated the successful transfection and the expression of the azurin-HA protein. Conspicuous apoptosis of the transfected cells was detected by flow cytometry (FCM) and the DNA ladder test. The apoptosis rate reached 64.3 ± 13.1%. The transcriptional levels of the Bax and p53 genes increased significantly in U2OS cells transfected with pcDNA3.1(+)/azurin, but the Bcl-2 mRNA level decreased. There was no difference in the levels of Bcl-xl mRNA and Survivin mRNA. We propose that the transfection of the recombinant plasmid pcDNA3.1(+)/azurin can significantly induce apoptosis in U2OS cells. This is closely associated with the up-regulation of the transcriptional level of the Bax and p53 genes, and the down-regulation of that of the Bcl-2 gene.
Keywords	Azurin, Transfection, Osteosarcoma, Apoptosis
Adress and Contact Informations	¹ Department of Orthopedics, Second Affiliated Hospital, Medical College, Zhejiang University, 88 Jie Fang Road, Hangzhou, 310009, Zhejiang, P.R. China, ²Department of Microbiology, Medical College, Zhejiang University, P.R. China, ³Institute of Cancer Research, Second Affiliated Hospital, Medical College, Zhejiang University, P.R. China * Author for correspondence; e-mail: dishengy44@sohu.com, fax: +86-571-8702-2776

DOI: 10.2478/s11658-007-0013-2 Volume 12 (2007) pp 422 - 434
Title	ENGRAFTING FETAL LIVER CELLS INTO MULTIPLE TISSUES OF HEALTHY ADULT MICE WITHOUT THE USE OF IMMUNOSUPPRESSANTS
Authors	Adas Darinskas¹, Renata Gasparaviciute², Mantas Malisauskas³, Kristina Wilhelm³, Jurij A. Kozhevnikov⁴, Evaldas Liutkevicius⁵, Audrone Pilinkiene⁵ and Ludmilla A. Morozova-Roche³*
Abstract	We have shown the fetal liver cell engraftments into multiple tissues of adult healthy mice, achieved without suppressing the animals’ immune systems. Fetal cells from the livers of male C57Bl/6J Black lineage mice at day 13 to 15 of gestation were injected intravenously into female adult CC57W/MY White mice. The grafting was evaluated by Y-chromosome-specific PCR, cytometric analysis of fluorescently stained donor cells, and histological analysis. All the methods consistently showed the presence of multiple engraftments randomly distributed through the various organs of the recipients. After 60 days, the grafts still constituted 0.1 to 2.75% of the tissues. The grafted cells did not change their appearance in any of the organs except the brain, where they became enlarged. Inflammatory reactions were not detected in any of the histological preparations. The frequency of engraftments was higher in the liver, indicating that similarity between the donor and recipient cells facilitates engraftment. The high inherent plasticity of fetal liver cells underlies their ability to integrate into healthy recipient organs, which can be governed by environmental conditions and connections with neighboring cells rather than by the initial cellular developmental programs. The fact that fetal liver cells can be grafted into multiple tissues of healthy animals indicates that they can be used to replace the natural loss of cells in adult organisms.
Keywords	Fetal cells, Transplantation, Engraftment, Immune suppression
Adress and Contact Informations	¹ Laboratory of Immunopharmacology, Institute of Immunology, Vilnius University, Vilnius 08409, Lithuania, ²Department of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Vilnius, Lithuania. Vilnius 08412, Lithuania, ³Department of Medical Biochemistry and Biophysics, Umea University, Umea SE-90187, Sweden, ⁴Laboratory of Cell Biotechnology, the Sibirian Division of the Russian Academy of Sciences, Institute of Clinical Immunology, Novosibirsk, 630091, Russia, ⁵“Imunolita” JSC, Vilnius 08217, Lithuania * Author for correspondence; e-mail: Ludmilla.Morozova-Roche@medchem.umu.se, tel: +46-90-7865283, fax: +46-90-7869795

DOI: 10.2478/s11658-007-0014-1 Volume 12 (2007) pp 435 - 447
Title	THE CFTR-DERIVED PEPTIDES AS A MODEL OF SEQUENCE- SPECIFIC PROTEIN AGGREGATION
Authors	Daniel Bąk^1,2*, Garry R. Cutting³ and Michał Milewski¹
Abstract	Protein aggregation is a hallmark of a growing group of pathologies known as conformational diseases. Although many native or mutated proteins are able to form aggregates, the exact amino acid sequences involved in the process of aggregation are known only in a few cases. Hence, there is a need for different model systems to expand our knowledge in this area. The so-called ag region was previously found to cause the aggregation of the C-terminal fragment of the cystic fibrosis transmembrane conductance regulator (CFTR). To investigate whether this specific amino acid sequence is able to induce protein aggregation irrespective of the amino acid context, we altered its position within the CFTR-derived C-terminal peptide and analyzed the localization of such modified peptides in transfected mammalian cells. Insertion of the ag region into a different amino acid background affected not only the overall level of intracellular protein aggregation, but also the morphology and subcellular localization of aggregates, suggesting that sequences other than the ag region can substantially influence the peptide’s behavior. Also, the introduction of a short dipeptide (His-Arg) motif, a crucial component of the ag region, into different locations within the C-terminus of CFTR lead to changes in the aggregation pattern that were less striking, although still statistically significant. Thus, our results indicate that even subtle alterations within the aggregating peptide can affect many different aspects of the aggregation process.
Keywords	Protein aggregation, Conformational diseases, CFTR, Site-directed mutagenesis
Adress and Contact Informations	¹ Laboratory of Cell Biology, Department of Medical Genetics, Institute of Mother and Child, Kasprzaka 17A, 01-211 Warsaw, Poland, ²Postgraduate School of Molecular Medicine, Żwirki i Wigury 61, 02-091 Warsaw, Poland, ³ Institute of Genetic Medicine, Johns Hopkins University School of Medicine, 733 N Broadway, Baltimore, MD 21287-3914, USA * Author for correspondence; e-mail: dbak@imid.med.pl, tel.: +48 22 3277 177, fax: +48 22 3277 200

DOI: 10.2478/s11658-007-0017-y Volume 12 (2007) pp 448 - 456
Title	THE EFFECT OF GROWTH MEDIUM ON THE ANTIOXIDANT DEFENSE OF Saccharomyces cerevisiae
Authors	Ewa Macierzyńska¹*, Agnieszka Grzelak¹ and Grzegorz Bartosz^1,2
Abstract	We compared the oxidation of dihydrorhodamine 123, glutathione contents and activities of superoxide dismutase (SOD) and catalase for three wild-type strains of Saccharomyces cerevisiae grown on media with different carbon sources. The rate of oxidation of dihydrorhodamine 123 was much higher in respiring cells grown on ethanol or glycerol media than in fermenting cells grown on glucose medium. The total SOD activity was highest on glycerol medium and lowest on ethanol medium, while the catalase activity was highest on glycerol medium. The sequence of glutathione content values was: glucose > ethanol > glycerol.
Keywords	Yeast, Saccharomyces cerevisiae, Reactive oxygen species, Superoxide dismutase, Catalase, Glutathione
Adress and Contact Informations	¹ Department of Molecular Biophysics, University of Łódź, Banacha 12/16, 90-237 Łódź, Poland, ²Department of Biochemistry and Cell Biology, University of Rzeszów, Cegielniana 12, 35-595 Rzeszów, Poland * Author for correspondence; e-mail: ematrix@biol.uni.lodz.pl, tel./fax: +48-42-6354476

DOI: 10.2478/s11658-007-0018-x Volume 12 (2007) pp 457 – 472
Title	THE EFFECT OF TGF-ß1 AND Smad7 GENE TRANSFER ON THE PHENOTYPIC CHANGES OF RAT ALVEOLAR EPITHELIAL CELLS
Authors	Guo-Ping Xu, Qing-Quan Li, Xi-Xi Cao, Qi Chen, Zhong-Hua Zhao, Zi-Qiang Diao and Zu-De Xu*
Abstract	The aim of this study was to investigate whether transforming growth factor-ß1 (TGF-ß1) could induce alveolar epithelial-mesenchymal transition (EMT) in vitro, and whether Smad7 gene transfer could block this transition. We also aimed to elucidate the possible mechanisms of these processes. The Smad7 gene was transfected to the rat type II alveolar epithelial cell line (RLE-6TN). Expression of the EMT-associated markers was assayed by Western Blot and Real-time PCR. Morphological alterations were examined via phase-contrast microscope and fluorescence microscope, while ultrastructural changes were examined via electron microscope. TGF-ß1 treatment induced a fibrotic phenotype of RLE-6TN with increased expression of fibronectin (FN), α-smooth muscle actin (α-SMA) and vimentin, and decreased expression of E-cadherin (E-cad) and cytokeratin19 (CK19). After transfecting the RLE-6TN with the Smad7 gene, the expression of the mesenchymal markers was downregulated while that of the epithelial markers was upregulated. TGF-ß1 treatment for 48 h resulted in the separation of RLE-6TN from one another and a change into elongated, myofibroblast-like cells. After the RLE-6TN had been transfected with the Smad7 gene, TGF-ß1 treatment had no effect on the morphology of the RLE-6TN. TGF-ß1 treatment for 48 h resulted in an abundant expression of α-SMA in the RLE-6TN. If the RLE-6TN were transfected with the Smad7 gene, TGF-ß1 treatment for 48 h could only induce a low level of α-SMA expression. Furthermore, TGF-ß1 treatment for 12 h resulted in the degeneration and swelling of the osmiophilic multilamellar bodies, which were the markers of type II alveolar epithelial cells. TGF-ß1 can induce alveolar epithelial- mesenchymal transition in vitro, which is dependent on the Smads signaling pathway to a certain extent. Overexpression of the Smad7 gene can partially block this process.
Keywords	Epithelial-mesenchymal transition, Gene transfer, Smad7, Transforming growth factor-ß1
Adress and Contact Informations	Department of Pathology, Shanghai Medical College, Fudan University, Shanghai 200032, China * Author for correspondence; e-mail: dxz_1028@163.com, tel: +86 021-54230267-2323

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