title: Analysis of TGF-β1- and drug-induced epithelial-mesenchymal transition in cultured alveolar epithelial cell line RLE/Abca3.
abstract: In this study, we examined the induction of epithelial-mesenchymal transition (EMT) by transforming growth factor (TGF)-β1 and drugs in genetically engineered type II alveolar epithelial cell line RLE/Abca3. Treatment of RLE/Abca3 cells with TGF-β1 induced marked changes in cell morphology from epithelial-like to elongated fibroblast-like morphology. With these morphological changes, mRNA expression of epithelial markers such as cytokeratin 19 (CK19) decreased, while that of mesenchymal markers such as α-smooth muscle actin (α-SMA) increased. TGF-β1 treatment also decreased the mRNA expression of Abca3, a type II cell marker, and formation of lamellar body structures. Interestingly, the effect of TGF-β1 on Abca3 mRNA expression was observed in RLE/Abca3 cells, but not in wild-type RLE-6TN, A549, and H441 cells. Treatment of RLE/Abca3 cells with bleomycin (BLM) and methotrexate (MTX) induced similar morphological and mRNA expression changes. In addition, the increase in α-SMA and the decrease in Abca3 mRNA expression by these drugs were observed only in RLE/Abca3 cells. These findings suggest that, like TGF-β1, BLM and MTX induce EMT in RLE/Abca3 cells, and RLE/Abca3 cells would be a good model to study drug-induced EMT. The effect of pirfenidone, an antifibrotic and anti-inflammatory drug, on EMT induced by TGF-β1 was also discussed.
authors.affiliation: Department of Pharmaceutics and Therapeutics, Graduate School of Biomedical & Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan. Electronic address: takanom@hiroshima-u.ac.jp.
authors.forename: Mikihisa
authors.initials: M
authors.lastname: Takano
authors.affiliation: Department of Pharmaceutics and Therapeutics, Graduate School of Biomedical & Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Chieko
authors.initials: C
authors.lastname: Yamamoto
authors.affiliation: Department of Pharmaceutics and Therapeutics, Graduate School of Biomedical & Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Koki
authors.initials: K
authors.lastname: Yamaguchi
authors.affiliation: Department of Pharmaceutics and Therapeutics, Graduate School of Biomedical & Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Kawami
authors.affiliation: Department of Pharmaceutics and Therapeutics, Graduate School of Biomedical & Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Ryoko
authors.initials: R
authors.lastname: Yumoto
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D000199:Actins / Q000378:metabolism; D000818:Animals; D001761:Bleomycin / Q000494:pharmacology*; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D006801:Humans; D053539:Keratin-19 / Q000378:metabolism; D008727:Methotrexate / Q000494:pharmacology*; D018613:Microscopy, Confocal; D011650:Pulmonary Alveoli / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D011728:Pyridones / Q000494:pharmacology; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*
chemical_list: C441524:ABCA3 protein, human; C541116:ACTA2 protein, human; D018528:ATP-Binding Cassette Transporters; D000199:Actins; D053539:Keratin-19; D011728:Pyridones; D053773:Transforming Growth Factor beta1; D001761:Bleomycin; C093844:pirfenidone; D008727:Methotrexate
keywords: Alveolar epithelial cells; Bleomycin; Epithelial–mesenchymal transition; Methotrexate; Pulmonary fibrosis; TGF-β1