Methylation Of Promoter In RASSF1A Gene And Its Impact On Expression In Gynecological Malignancy Cell Lines

Partâ… : Expression of the RASSF1A gene and methylation in its promoter in gynecological malignancy cell linesObjective To study the DNA methylation status of RASSF1A gene promoter region in gynecological malignancy cell lines and its correlation with RASSF1A gene expression.Methods Methylation-specific PCR were performed to detect DNA methylation status of RASSF1A gene in its promoter region in cell lines: A2780, Anglne, Caov3, COC1, SKOV3, HEC-1B, Ihsikawa, Hela, Siha. The mRNA expression of RASSF1A gene was detected by RT-PCR.Results A2780, Anglne, Caov3, SKOV3, HEC-1B, Ishikawa cell lines showed DNA hypermethylation in RASSF1A gene promoter region, and partial DNA methylation was identified in COC1, Hela and Sina cells; Meanwhile extremely low levels of RASSF1A expression was found in A2780, Anglne, Caov3, SKOV3, HEC-1B, Ishikawa cell lines, and slightly higher levels of RASSF1A expression in COC1, Hela and Siha cells.Conclusion Methylation of RASSF1A gene promoter region is hight in gynecological malignancy cell lines and is one of the important mechanisms for the loss of expression of this gene. Partâ…¡: Effect of 5-2aza-CdR on methylation status of RASSF1A gene promoter region and on its expression in ovarian cancer cell line SKOV3Objective To detect the effect of 5-2aza-CdR on methylation of RASSF1A gene and on its expression; meantime to check the invasion ability and growth activation of ovarian cancer cell line SKOV3 under 5-2aza-CdR treatment.Methods Methylation-specific PCR(MSP) and RT-PCR were used to detect the methylation status and mRNA expression of RASSF1A gene in ovarian cacer cell SKOV3 before and after the treatment with 5-2aza-CdR; The growth activity of cells was determined by MTT assay after the 5-2aza-CdR treatment and the invasion ability of cells was tested by Transwell experiments before and after the treatment with 5-2aza-CdR.Result1. After treatment with 5-2aza-CdR, accompanied with the decrease of methylated levels and the increase of unmethylated levels in its promoter region, the RASSF1A gene expression in ovarian cancer cell line SKOV3 was enhanced.2. The activation of the cell growth and invasion ability of SKOV3 after treatment with 5-2aza-CdR were obviously declined.Conclusion 5-2aza-CdR may effectively cause demethylation of DNA of RASSF1A gene and inhibit the growth and invasion of cancer cells by restoring the gene transcription and expression.