肝癌细胞系中Runx3基因表达及启动子区异常甲基化分析

Expression and Hypermethylation of Runx3 gene in Promoter Region with Hepatocellular Carcinoma Cell Lines

目的:通过检测6种肝癌细胞中Runx3基因异常甲基化状态及表达情况,探讨药物5-aza-2’-de-oxycytidine(decitabine)激活Runx3基因重新表达的能力及对肝癌细胞生长的影响。方法:采用DNA甲基化特异性PCR(MSP)技术分别对6种肝细胞癌细胞系Runx3基因启动子区域甲基化状态进行检测,同时采用逆转录-聚合酶链反应(RT-PCR)检测5种肝癌细胞中Runx3的表达情况及药物5-aza-2’-de-oxycytidine处理其中3种肝癌细胞前后Runx3表达变化。结果:6种肝癌细胞系中,有3种细胞Runx3基因启动子区域存在甲基化异常,药物5-aza-2’-deoxycytidine处理3种肝癌细胞后,Runx3基因明显表达或表达活性增强。结论:启动子区异常甲基化是导致Runx3基因失活的主要原因之一,与肝细胞癌发生早期密切相关,可作为肝细胞癌早期诊断的分子标记物及分子治疗的靶点。

Objective:To detect the status of promoter hypermethylation and expression of Runx3 gene in six hepatocellular carcinoma(HCC) cell lines and investigate the effect of 5-aza-2'-deoxycytidine(decitabine) on cell growth and the ability of reactivation of Runx3 gene.Methods:The methylation status of Runx3 was examined by methylation-specific polymerase chain reaction(MSP).Meanwhile,RT-PCR was used to detect expression of Runx3 gene in six HCC cell lines and three cell lines that were treated with 5-aza-2'-deoxycytidine.Results:3 of 6 HCC cell lines showed hypermethylation in the promoter region of Runx3 gene.Demethylating agent 5-aza-2'-deoxycytidine induced reactivation and more potent expression of Runx3 gene in the three cell lines.Conclusion:High frequent hypermethylation was one of reasons which induced Runx3 gene inactivation in human hepatocellular carcinoma,hypermethylation of Runx3 promoter may be correlated with the early phase of hepatocellular carcinoma,Runx3 gene can be utilized as a molecular diagnostic marker and therapy target of hepatocellular carcinoma.