5-Aza-CdR体外诱导人肺癌细胞株p16基因去甲基化

5-Aza-CdR Induced Demethylation of p16 Gene in Human Lung Cancer Cell Lines

目的观察人肺癌细胞株H719p16基因启动子区甲基化状态,并探讨去甲基化制剂5-氮杂-2′-脱氧胞苷(5-Aza-CdR)诱导H719细胞株高甲基化失活的pl6基因重新表达的可能性。方法MSP法检测体外培养的人肺癌细胞株p16基因启动子区甲基化状态,并应用不同浓度的5-Aza-CdR处理人肺癌细胞株H719,MSP法检测用药后细胞中p16基因的甲基化状态,并用RT-PCR方法及Western Blot方法检测用药前后细胞中p16基因mRNA和蛋白水平变化。结果p16基因在人肺癌细胞株H719中启动子区呈甲基化状态,经过5-Aza-CdR处理后,p16基因启动子区呈去甲基化状态,并且其mRNA及蛋白恢复表达。结论启动子区异常甲基化是人肺癌细胞株p16基因失活的可能原因,5-Aza-CdR能逆转p16基因甲基化状态。

Objective To investigate the methylation status of p16 gene promoter in human lung cancer cell line H719 and to explore the possibility of re-expression of the hypermethylated and silenced p16 gene.Methods Methylation-specific PCP(MSP) was used to detect methylation state of p16 gene promoter,and H719 were treated with different concentrations of DNA methyltransferase inhibitor 5-Aza-CdR.The re-expression of p16 mRNA and protein were detected by RT-PCR and Western blot before and after treatment with 5-Aza-...