EGCG抑制卵巢癌细胞RASSF1A基因甲基化并上调其表达  

Effect of EGCG on Methylation and Expression of RASSF1A in Ovarian Cancer Cell Line A2780

作　　者：申清香[1] 魏付桥[1] 

机构地区：[1]南华大学第二附属医院妇产科,湖南衡阳421001

出　　处：《中南医学科学杂志》2011年第3期273-276,共4页Medical Science Journal of Central South China

摘　　要：目的探讨表没食子儿茶素没食子酸酯（EGCG）对卵巢癌细胞系A2780Ras相关区域家族1基因（RASSFl）甲基化及其表达的影响。方法体外培养A2780细胞，用0、10、30和50μmol／LEGCG与A2780细胞孵育24-72h，采用甲基化特异性PCR（MSP）检测RASSFlA基因启动子区域的甲基化改变情况；提取EGCG处理后A2780细胞的RNA，用逆转录PCR（RT-PCR）检测RASSFlA的表达水平。结果静息状态下，MSP结果显示A2780细胞可检测出RASSFlA基因启动子区域有明显的甲基化条带，当与50μmol／LEGCG孵育72h后，甲基化水平有所减弱，而非甲基化基因增多。RT-PCR显示EGCG呈剂量和时间依赖性地促进A2780细胞RASSFlA基因mRNA和蛋白表达。结论EGCG能使RASSFlA基因去甲基化，并上调其非甲基化基因和蛋白的表达水平，这可能是其抗肿瘤的奄耍机制。Objective To investigate the effect of epigallocatechin-3-gallate（EGCG）on RASSF1A methylation and protein expression in a cell line of A2780. Methods The cultured A2780 cells were incubated with EGCG for 24 - 76 h. After that, DNA was extracted, and the methylation in RASSF1A promoter region was determined by methylation-specific PCR（MSP）. The mRNA and protein level of RASSF1A was detected by RT-PCR and Western blot. Results PCR resuits showed non-methylation band and methylation band was detected when A2780 cells was under quiescent condition. When the cells were incubated with 50 jxmol/L EGCG, the methylation band level was decreased, and the non-methylated DNA was increased. RT-PCR and Western blot also showed the mRNA level of RASSF1A was increased in dose and timedependent manner. Conclusion Demethylation of RASSF1A and increasing its mRNA and protein induced by EGCG mav be one of the mechanisms of its antineoolastic activity.

关 键 词：卵巢癌细胞 表没食子儿茶素没食子酸酯 Ras相关区域家族1基因 

分 类 号：R737.31[医药卫生—肿瘤]