Title: Methylation status of promoter of RUNX3 gene in epithelial ovarian cancer cell lines
Abstract: Objective To investigate the methylation status of RUNX3 promoter region in the epithelial ovarian cancer cell line and effect of 5-aza-2'-deoxycytidine on the proliferation and apoptosis of ovarian cancer cells and the expression of tumor suppressor gene RUNX3.Methods Methylation status of RUNX3 promoter region was assayed in human ovarian cancer cell line 3AO,KOV3 and normal ovarian epithelial cell (NOEC) by methylation-specific PCR(MSP).Ovarian cancer cell line was treated with 5-aza-2'-deoxycytidine (0.5,5,50 μmol/L),a specific demethylating agent for 3 d,and then cultured in RPMI 1640 medium for 5 d.The growth of 3AO and SKOV3 cells was observed by MTT assay before and after 5-aza-2'-deoxycytidine treatment respectively.The expression of RUNX3 mRNA was observed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).The apoptosis of 3AO and SKOV3 cells was analyzed by flow cytometry.Results The methylation of RUNX3 promoter was found in both 3AO and SKOV3 but not in NOEC.Ovarian cancer cells treated with 5-aza-2'-deoxycytidine displayed a slowed growth in comparison with the control cells,and methylation was induced or completely reversed and 5-aza-2'-deoxycytidine as a methylase inhibitor can reverse,recovery and reinforce RUNX3 mRNA expression their mRNA expression was increased differently.Conclusions 5-aza-2'-deoxycytidine as a methylase inhibitor can reverse,recovery and reinforce RUNX3 mRNA expression of ovarian cell line 3AO and SKOV3,repress cancer cell growth and induce its apoptosis.
Publication Year: 2009
Publication Date: 2009-01-01
Language: en
Type: article
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