摘要
目的:探讨甲基化转移酶抑制剂5-氮杂-2’-脱氧胞苷(5-Aza-2’-deoxycytidine,5-Aza-CdR)对人子宫内膜癌(Humanendometrial cancer,HEC)-1-B细胞增殖和凋亡的作用及其可能机制。方法:应用不同浓度的5-Aza-CdR处理HEC-1-B细胞,甲基化特异PCR(Methylation-specific polymerase chain reaction,MSP)法检测处理前后RAS相关域家族1A(RAS association do-main family 1A,RASSF1A)基因甲基化状态,应用RT-PCR方法检测RASSF1A基因mRNA表达,MTT法检测细胞增殖,流式细胞术检测细胞凋亡率。结果:HEC-1-B细胞RASSF1A基因启动子区呈高甲基化状态,经过5-Aza-CdR处理后,RASSF1A基因启动子区呈去甲基化状态,其mRNA重新表达,细胞生长受到抑制,细胞凋亡增加。结论:5-Aza-CdR能够逆转RASSF1A基因甲基化状态,调控RASSF1A基因表达,并能有效地抑制HEC-1-B细胞的增殖和诱导细胞调亡。
Objective：To investigate the effects of 5-aza-2＇-deoxycytidine（5-Aza-CdR） on the proliferation and apoptosis in human endometrial cancer（HEC）HEC-1-B cells and the possible mechanism.Methods：HEC-1-B cells were treated with different concentrations of DNA methytransferase inhibitor 5-Aza-CdR.Methylation-specific polymerase chain reaction（MSP） was used to detect promoter methylation status of RAS association domain family 1A（RASSF1A） gene and RT-PCR was used to detect re-expression of mRNA.Furthermore,cell growth was estimated by MTT assay and cell apoptosis rate was estimated by flow cytometry.Results：Promoter hypermethylation of the RASSF1A gene was detected in HEC-1-B cells.After treatment with 5-Aza-CdR,the promoter region of the RASSF1A gene exhibited a demethylation status,and RASSF1A gene was re-expressed.Meanwhile,the cellular growth activity decreased and cell apoptosis increased.Conclusions：The 5-Aza-CdR could reverse methylation of RASSF1A gene to regulate the expression of RASSF1A gene,and could effectively inhibit the cellular proliferation and induce apoptosis of endometrial carcinoma cells.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2012年第6期525-527,共3页
Journal of Chongqing Medical University
