Development of oral epithelial cell line ROE2 with differentiation potential from transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene

Yoshiaki Tabuchi, Shigehito Wada, Mika Ikegame, Ayako Kariya, Yukihiro Furusawa, Nobuhiko Hoshi, Tatsuya Yunoki, Nobuo Suzuki, Ichiro Takasaki, Takashi Kondo, Yoshihisa Suzuki

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Abstract

We have developed an immortalized oral epithelial cell line, ROE2, from fetal transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene. The cells grew continuously at either a permissive temperature of 33°C or an intermediate temperature of 37°C. At the nonpermissive temperature of 39°C, on the other hand, growth decreased significantly, and the Sub-G1 phase of the cell cycle increased, indicating that the cells undergo apoptosis at a nonpermissive temperature. Histological and immunocytochemical analyses revealed that ROE2 cells at 37°C had a stratified epithelial-like morphology and expressed cytokeratins Krt4 and Krt13, marker proteins for oral nonkeratinized epithelial cells. Global-scale comprehensive microarray analysis, coupled with bioinformatics tools, demonstrated a significant gene network that was obtained from the upregulated genes. The gene network contained 16 genes, including Cdkn1a, Fos, Krt13, and Prdm1, and was associated mainly with the biological process of skin development in the category of biological functions, organ development. These four genes were validated by quantitative real-time polymerase chain reaction, and the results were nearly consistent with the microarray data. It is therefore anticipated that this cell line will be useful as an in vitro model for studies such as physiological functions, as well as for gene expression in oral epithelial cells.

Original languageEnglish
Pages (from-to)31-44
Number of pages14
JournalExperimental Animals
Volume63
Issue number1
DOIs
Publication statusPublished - 2014 Feb 11

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Keywords

  • Differentiation
  • Gene network
  • Oral epithelial cell
  • Temperature-sensitive simian virus 40 large T-antigen

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Biochemistry, Genetics and Molecular Biology(all)
  • veterinary(all)

Cite this

Tabuchi, Y., Wada, S., Ikegame, M., Kariya, A., Furusawa, Y., Hoshi, N., Yunoki, T., Suzuki, N., Takasaki, I., Kondo, T., & Suzuki, Y. (2014). Development of oral epithelial cell line ROE2 with differentiation potential from transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene. Experimental Animals, 63(1), 31-44. https://doi.org/10.1538/expanim.63.31