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Archive > Volume 24 Issue 6 > 2017,24(6):627-631. DOI:10.3872/j.issn.1007-385X.2017.06.009 Prev Next
Basic Research
5AzaCdR effects the methylation and expression of ERCC1 gene in ovarian cancer cell via DNMT1
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Abstract:
Objective:Objective: To investigate the effects of 5AzaCdR on expression of excision repair cross complementation group 1 (ERCC1) in ovarian cancer cell line SKOV3 and its possible mechanism.Methods: shRNA that specifically targeting DNA methyltransferase 1 (DNMT1) was constructed and transfected into SKOV3 cells. Western blotting was applied to determine the expressions of DNMT1 and ERCC1 in SKOV3 cells; Ovarian cancer cell line SKOV3 was treated with 5AzaCdR of various concentrations for different time courses, and Western blotting was used to determine the protein changes of DNMT1 and ERCC1 before and after the treatment; and the methylation level of ERCC1 promoter was tested by Sodium Bisulfite method. Results: After being treated with 5AzaCdR (at the concentrations of 0.5, 1.0, 20, 4.0 μmol/L), the expression of DNMT1 protein in SKOV3 cells was decreased in a dosedependent manner while the ERCC1 protein was increased in a timedependent manner; With the treatment of 5AzaCdR (concentration of 1.0 μmol/L) for 12, 24 and 36 h, the expression of DNMT1 protein was decreased and the ERCC1 protein was increased in SKOV3 cells, and the expression both changed in a timedependent manner. Sodium Bisulfite method indicated that the methylation of ERCC1 promoter was at high level before the treatment; However, after the treatment of 5AzaCdR at concentration of 1.0 μmol/L, the promoter of ERCC1 showed demethylation．Conclusion: 5AzaCdR regulates the methylation of ERCC1 promoter and its expression in SKOV3 cells via.
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