Effect of the promoter methylation and the intervention by 5-aza-2'-deoxycytidine on RASSF1 A gene of breast cancer cell line MCF-7
AIM:To approach the effect of promoter methylation to RASSF1A gene of breast cancer cell line MCF-7 and 5-AzaCdR's impact on promoter methylation above.METHODS:Breast cancer cell lines MCF-7 were treated with 1,3,5,7 and 9 μmol/L 5-Aza-CdR respectively and MTT assay was used to detect the survival rate.Then cells were treated with 5 and 10 μmol/L 5Aza-CdR,the cell cycle and apoptosis rate were analyzed by flow cytometry and the methylation was examined by PCR using methylation specific primer(MSP),and the expression levels of mRNA of RASSF1A were detected by reverse transcription-polymerase chain reaction(RT-PCR).RESULTS:After 5-Aza-CdR treatment,the proliferation of breast caner cell line MCF-7 is inhibited,the number of G0/G1 cell was increased,and 5-AzaCdR blocked the cell cycle at G1/G0 phase,the apoptotic rate was also increased significantly in a concentration dependent manner.After 5-Aza-CdR treatment,methylation specific primer(MSP)detected RASSF1 A gene in breast cancer cell line generated methylation.RT-PCR detected mRNA of RASSFl A gene in breast cancer cell line recovered express.CONCLUSION:The promoter methylation of RASSF1 A gene in breast cancer cell line MCF-7 is the main reason inducing itself deactivation,while 5-Aza-CdR call reverse that promoter methylation to make RASSFl A gene express again.
NETL
NSTL
万方数据
