EGCG对黑素瘤A375细胞增殖及RASSF1A基因表达的影响

摘要

Objective: To assess the effect EGCG (Epigallocatechin gallate) on the proliferation of human melanoma cell line A375 and the expression of RASSF1A (Ras - assotiation domain family 1 A) gene. Methods: After human melanoma cell line A375 was treated with EGCG in different concentrations, the inhibition of the growth of A375 cell line measured with MTT method in different time interval, and the methylation of RASSF1A and the expression of RASSF1A mRNA gene detected by methylation - specific PCR (MS - PCR) and RT - PCR before and after the treatment. Results: The growth of A375 cells was inhibited when treated with a concentration of 5 ~ 35 μmol/L EGCG. The inhabitation was time - dependent. Methylaiton in the promoter region of RASSF1A gene was reversed from a high methylation status before the treatment with EGCG to a low methylation status after the treatment with EGCG. The degree of RASSF1A mRNA expression in the cells was positively correlated to the concentration of EGCG and the time of the treatment. Conclusion: EGCG can inhibit the growth of A375 cell line and reverse methylation status in the promoter region of RASSF1A resulting in re - expression of the gene and inhibition of the proliferation of the cells.%目的:评价表没食子儿茶素没食子酸酯(EGCG)对人黑素瘤A375细胞增殖及Ras相关区域家族1A(RASSF1A)基因表达的影响.方法:不同浓度的EGCG处理A375细胞后,分别于不同时间点用MTr法检测A375细胞的生长抑制率；并应用甲基化特异性PCR法以及荧光半定量PCR法检测EGCG作用前后细胞中RASSF1A甲基化及mRNA的表达.结果:EGCG浓度在5～30 μg/mL时抑制细胞生长并呈时间剂量依赖；作用后可使RASSF4A基因由原有的高甲基化变为低甲基化；细胞中RASSF1A基因的mRNA表达量与药物作用浓度和作用时间呈正相关.结论:EGCG可以抑制A375细胞的生长,并可通过逆转RASSF1A基因启动子区域甲基化状态,恢复该基因在细胞中的部分表达,从而达到抑制其增殖的作用.