申清香,魏付桥.EGCG抑制卵巢癌细胞RASSF1A基因甲基化并上调其表达.[J].中南医学科学杂志.,2011,39(3):273-276. |
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 |
投稿时间:2010-12-07 |
DOI: |
中文关键词: 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 |
英文关键词:epigallocatechin-3-gallate ovarian cancer ras-association domain family 1 |
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中文摘要: |
目的探讨表没食子儿茶素没食子酸酯(EGCG)对卵巢癌细胞系A2780 Ras相关区域家族1基因(RASSF1)甲基化及其表达的影响。方法体外培养A2780细胞,用0、10、30和50 μmol/L EGCG与A2780细胞孵育24~72 h,采用甲基化特异性PCR(MSP)检测RASSF1A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RASSF1A的表达水平。结果静息状态下,MSP结果显示A2780细胞可检测出RASSF1A基因启动子区域有明显的甲基化条带,当与50 μmol/L EGCG孵育72 h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RASSF1A基因mRNA和蛋白表达。结论EGCG能使RASSF1A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重要机制。 |
英文摘要: |
ObjectiveTo investigate the effect of epigallocatechin-3-gallate(EGCG)on RASSF1A methylation and protein expression in a cell line of A2780.MethodsThe 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.ResultsPCR results showed non-methylation band and methylation band was detected when A2780 cells was under quiescent condition.When the cells were incubated with 50 μmol/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 time-dependent manner.ConclusionDemethylation of RASSF1A and increasing its mRNA and protein induced by EGCG may be one of the mechanisms of its antineoplastic activity. |
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