5-Aza-CdR对子宫内膜癌中RASSF1A基因甲基化的作用  

The Role of 5-Aza-CdR on Methylation of Promoter in RASSF1A Gene in Endometrial Carcinoma

作　　者：黄丽萍 陈晨[2] 王雪萍[2] 刘辉[2] 

机构地区：[1]山东省德州市市立医院妇产科,德州253012 [2]四川大学华西第二医院妇产科,成都610041

出　　处：《四川大学学报（医学版）》2015年第3期380-383,共4页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅科技支撑计划项目(No.2010SZ0107)资助

摘　　要：目的探讨去甲基化药物5-氮杂-2’-脱氧脱苷(5-Aza-CdR)对子宫内膜癌中Ras相关区域家族1A(RASSF1A)基因甲基化的作用。方法取子宫内膜癌细胞株HEC-1-B细胞随机分组,用不同浓度(1.0×10-6,3.0×10-6,10.0×10-6 mol/L)的去甲基化药物5-Aza-CdR处理后,采用甲基化特异性PCR、实时定量PCR、Western blot、TUNEL等技术,分析空白对照组及实验各组子宫内膜癌细胞株HEC-1-B中RASSF1A基因启动子区域甲基化情况、mRNA表达水平、蛋白表达和细胞凋亡的情况。结果人子宫内膜癌细胞株HEC-1-B细胞中RASSF1A基因启动子区存在高甲基化,mRNA、蛋白低表达,以及细胞凋亡失控。不同浓度的5-Aza-CdR可逆转RASSF1A基因高甲基化状态,恢复mRNA、蛋白表达量,并可抑制HEC-1-B细胞的生长及诱导凋亡。结论子宫内膜癌异常甲基化RASSF1A基因作为治疗靶点,去甲基化药物5-Aza-CdR是基因治疗的有效途径。Objective To explore the effect of demethylating drug 5-Aza-2′-deoxycytidine（5-Aza-CdR）on methtylation status of the Ras-association domain family1 A gene（RASSF1A）in human endometrial carcinoma.Methods Randomly assign the human endometrial carcinoma cell line HEC-1-B into groups and use demethylating drug 5-Aza-CdR of different concentration to treat them.Then Methylation-specific polymerase chain reaction（MSP）,real-time PCR,Western blot,TUNEL technology were used to analyze methylation status of RASSF1 A promoter CpG islands,RASSF1 A mRNA expression,RASSF1 Aprotein expression and apoptosis of HEC-1-B cell.Results High DNA methylation in RASSF1 A gene promoter region,low RASSF1 A mRNA level and protein expression and out of control of human endometrial carcinoma cell HEC-1-B apoptosis were observed.5-Aza-CdR of different concentration could reverse RASSF1 A gene＇s methylation status,recover the expression of mRNA and protein,and control the growth of HEC-1-B by inducing apoptosis.Conclusion Aberrant methylation of RASSF1 A in endometrial cancer as a therapeutic target,demethylating agent 5-Aza-CdR could be an effective way of gene therapy.

关 键 词：RASSF1A基因 甲基化 子宫内膜癌 

分 类 号：R737.33[医药卫生—肿瘤]