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T. Kawakita, S. Shimmura, N. Okada, S. Yoshida, S. Kawasaki, K. Tsubota; Epigenetic Regulation of Cytokeratin12 and Pax6 Expression in Mouse Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):6374.
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© ARVO (1962-2015); The Authors (2016-present)
Cytokeratin 12 is generally used as corneal epithelial specific marker. In human corneal epithelial cell lines such as SV40-induced cells, it has been reported that cytokeratin 12 did not expressed. Primary corneal epithelial cells also did not expressed after several passages. This regulation of Cytokeratin 12 has reported to be related to epigenetic regulation. The aim of this study was to determine whether Cytokeratin 12 of non-genetically induced mouse corneal epithelial cell line was also regulated epigenetically, and to further analyzing whether pax6 was also regulated epigenetically or not.
Mouse corneal epithelial cell lines (passage 28 and 182) methylation status of Cytokeratin 12 genome was analyzed as previously reported by Kawasaki et al in ARVO annual meeting. Mouse tissue of cornea, conjunctivae, skin and oral mucosa was used as control. Trichostatin A (TSA) 10-1000 nM and 5-Azacytidine (5-Aza) 10-1000 nM n was applied to cells, and Cytokeratin 12 and pax6 expression were confirmed by RT-PCR and immunostaining.
Cytokeratin 12 genome maintained low- methylation pattern in corneal tissue to compare with mouse cell lines (in both passage 28 and 182), conjunctivae, skin and oral mucosa. Cytokertain 12 and pax6 expression was recovered by Trichostatin A (TSA) and 5-Azacytidine in RNA level both in dose dependently to compare with control, but immunostaining showed low expression level.
Cytokeratin 12 expression was epigetically regulated in mouse corneal epithelial cell lines. TSA and 5-Aza could recovered Cytokeratin 12 and pax6 expression in RNA level.
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