组蛋白甲基转移酶G9a抑制剂联合索拉菲尼对肾癌细胞增殖能力的影响

Effect of Histone Methyltransferase G9a Inhibitor Combined with Sorafenib on Proliferation of Renal Cell Carcinoma

目的 G9a作为组蛋白甲基化转移酶,探究其特异性抑制剂 BIX-01294与经典抗癌药物索拉菲尼联合使用对于肾癌细胞增殖能力的影响。方法 BIX-01294与索拉菲尼各单独或联合作用于A498肾癌细胞时,通过实时荧光定量和蛋白免疫印记法检测G9a的mRNA和蛋白的表达水平,蛋白免疫印记法检测G9a蛋白以及其下游基因H3K9me2的蛋白表达水平,克隆形成实验测定A498细胞的增殖能力,流式细胞术法检测A498细胞的凋亡情况。结果 BIX-01294与索拉菲尼单独使用以及联合用药都能引起G9a蛋白表达水平的下调,同时引起A498细胞增殖能力降低,凋亡水平上调,两者联合使用效果更加明显。结论 BIX-01294 以及索拉菲尼均可引起肾癌细胞G9a mRNA以及蛋白的表达水平下调,两种药物联合使用效果最好,可明显降低细胞的增殖能力,同时细胞凋亡程度更为明显,为肾癌患者的化疗治疗、用药方法以及后续的机制研究提供了全新的参考。

OBJECTIVE To investigate the inhibitory effect of combination of histone methylation enzyme G9a specific inhibitor BIX-01294 and sorafenib against renal cell carcinoma. METHODS The mRNA expression level of G9a was detected by RT-PCR, the protein expression level of G9a and its target H3K9me2 were detected by WB in A498 cells. Colony formation assay was used to identify the effect of drug on cell proliferation and migration. Cell apoptosis was detected by flow cytometry. RESULTS G9a expression level can be reduced by BIX-01294 and sorafenib alone and double using of BIX-01294 and sorafenib could down regulate G9a gene expression effectively. Proliferation and migration of A498 cells were decreased, the apoptosis rate was increased present with drugs treatment. CONCLUSION G9a expression level can be down-regulated by either BIX-01294 or sorafenib in renal cancer cells, combined using can get a marked effect. Combining using of BIX-01294 and sorafenib can reduce cell proliferation and migration significantly, can promote cell apoptosis obviously. This research provides us a reference for renal cancer treatment, and the method of administration, the next-step mechanism research.