顺铂通过p19/p21途径诱导肝癌细胞衰老的探讨

Cisplatin induces senescence of hepatoma cells via p19/p21 pathway

目的探讨顺铂对肝癌细胞衰老的诱导作用。方法使用2μg/ml顺铂处理人肝癌细胞系HepG2,采用MTS法检测细胞增殖、流式细胞系检测细胞周期变化、RT-PCR和Western blot检测衰老相关基因P19、p21表达量、免疫荧光法检测P21表达情况;裸鼠移植人肝癌模型腹腔注射相应药物,检测肝癌生长转移及P21蛋白表达。结果随着顺铂处理时间延长,顺铂组增殖率较对照组降低,差异有统计学意义(t=8.958,P<0.05)。流式细胞检测显示,顺铂组和对照组G2期细胞分别为19.90%±0.42%、12.57%±0.84%,细胞停滞于G2期(t=14.415,P<0.05)。顺铂组与对照组p19基因相对表达量分别为(2.23±0.05)、(1.00±0.02);p21基因相对表达量分别为(3.26±0.11)、(1.00±0.02),pl9及p21表达均升高(分别t=43.750,56.541,均P<0.05);顺铂组裸鼠瘤体体积较对照组缩小,p21蛋白表达水平较对照组增加(P<0.05)。结论顺铂可通过p19/p21相关途径诱导肝癌细胞衰老。

Objective To investigate the induction mechanism of cisplatin on hepatoma senescence.Methods 2μ/ml cisplatin was applied to affect hepatocellular carcinoma cell line HepG2.Cell proliferation of tumor cells was detected by MTS method.The cell cycle was detected by flow cytometry.The RT-PCR and Western blot were used to detect the expression of senescence regulation genes,p19 and p21.The expression of p21 was detected by cell immunofluorescence.Nude mouse model of human hepatocellular carcinoma was intraperitoneally injected with specific drug to measure the growth,metastasis of tomor and expression of p21 protein.Results Cell proliferation rate in cisplatin group was lower than that of the control group(t=8.958,P<0.05).Flow cytometry showed the percentage of G2 phase were 19.90%±0.42%,12.57%±0.84%in the cisplatin and control group,respectively.Cells were arrested in G2 phase(t=14.415,P<0.05).In the cisplatin group and control group,the relative expression of pl9 were(2.23±0.05),(1.00±0.02);the relative expression of p21 were(3.26±0.11),(1.00±0.02).The expression of p19 and p21 increased(respectively t=43.750,56.541,all P<0.05).The tumor size of cisplatin group was smaller than that of the control group,and the expression level of p21 protein was higher than that of the control group(P<0.05).Conclusion Cisplatin can induce senescence of hepatocellular carcinoma cells through the p19/p21 pathway.