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论文摘要

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, 王雪萍等

Author:HUANG Li-ping, CHEN Chen, WANG Xue-ping. et al

收稿日期:          年卷(期)页码:2015,46(3):380-383

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:RASSF1A基因 甲基化 子宫内膜癌

Key words:RASSF1A gene Methylation Endometrial cancer

基金项目:

中文摘要

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。

英文摘要

Objective To explore the effect of demethylating drug 5-Aza-2′-deoxycytidine (5-Aza-CdR) on methtylation status of the Ras-association domain family1A 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 RASSF1A promoter CpG islands,RASSF1A mRNA expression, RASSF1A protein expression and apoptosis of HEC-1-B cell. Results High DNA methylation in RASSF1A gene promoter region, low RASSF1A 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 RASSF1A 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 RASSF1A in endometrial cancer as a therapeutic target, demethylating agent 5-Aza-CdR could be an effective way of gene therapy.

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