Development and Characterization of Conditionally Immortalized Gastric Epithelial Cell Lines from Transgenic Rats Harboring Temperature-Sensitive Simian Virus 40 Large T-antigen Gene

Cell Structure and Function

Shinji Asano⁴  Yuko Arai³  Yoshiaki Tabuchi⁴  Masuo Obinata¹  Hiroki Shioya³  Masatsugu Ueda²  Shoichiro Ohta⁵  Noriaki Takeguchi³  Ri-ichi Takahashi² 

[1] Institute of Development, Aging and Cancer, Tohoku University;YS New Technology Institute Inc.;Faculty of Pharmaceutical Science;Molecular Genetics Research Center;Faculty of Medicine, Toyama Medical and Pharmaceutical University

关键词: temperature-sensitive simian virus 40 large T-antigen;    transgenic rat;    gastric epithelial cell lines;    cathepsin E;    prostaglandin E2;   

DOI  :  10.1247/csf.27.71

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(45)Cited-By(8)Conditionally immortalized gastric epithelial cell lines were established from transgenic rats harboring temperature-sensitive simian virus 40 (tsSV40) large T-antigen gene. Gastric mucosal cells and epithelial tissues isolated from the stomach of the transgenic rats were cultured at permissive temperature (33°C), and proliferative cells were cloned by colony formation. Six cell lines (designated as RGE1-01, RGE1-02, RGE1-03, RGE1-21, RGE1-22 and RGE2-01) showing epithelial-like morphology have been established. All cells grew at 33°C, but did not at nonpermissive temperature (39°C). High expression level of large T-antigen in the nuclei was observed at 33°C, whereas the expression level was gradually decreased in a time-dependent manner at 39°C. These results suggest that the temperature-sensitive growth characteristics arise as a result of a function of the tsSV40 large T-antigen. None of the cell lines were transformed as judged by anchorage-independent growth assay. Immunocytochemical findings indicated that all cells expressed epithelial cell markers including cytoskeletal (cytokeratin and actin), basement membrane (laminin and collagen type IV) and junctional complex (ZO-1 and desmoplakin I+II) proteins at 33°C. All cells expressed mRNA of cathepsin E, a pit cell marker. Moreover, transepithelial resistance was observed between apical and basolateral sides in the cells. RGE1-22 cells produced prostaglandin E2. Levels of mRNA for cathepsin E, transepithelial resistance and prostaglandin E2 were influenced by the nonpermissive temperature. Thus, these conditionally immortalized gastric cell lines which preserve some epithelial cell characteristics will provide a useful in vitro model of gastric epithelium.

【 授权许可】

Unknown