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目的化疗药物能够通过诱导肿瘤细胞衰老从而发挥治疗效果。顺铂作为最常用的化疗药物能否诱导肝癌细胞发生加速性衰老目前尚不清楚。方法使用MTT法和克隆形成试验检测不同剂量顺铂对HepG2细胞增殖的影响;分别用流式细胞仪及衰老相关β-半乳糖苷酶染色检测细胞周期及衰老情况;RT-PCR检测TP53、P21及P19基因的mRNA表达水平,Western blotting检测P53和P21蛋白表达水平。结果顺铂能够诱导HepG2细胞出现不可逆的生长停滞及细胞周期阻滞。2.0μg/ml顺铂作用于HepG2后衰老相关β-半乳糖苷酶染色阳性,并呈现时间依赖性。在顺铂诱导衰老过程中,P19基因表达水平升高,在诱导48 h后达到峰值后逐渐下降,而P53和P21表达水平则持续升高。结论本研究提示顺铂能够诱导肝癌细胞出现衰老样表型,这一结果为进一步探讨其抗肝癌机制提供了基础。
The purpose of chemotherapy drugs can induce the tumor cells to senescence and thus play a therapeutic effect. Whether cisplatin as the most commonly used chemotherapeutic agent induces accelerated senescence in HCC cells is not clear. Methods MTT assay and colony formation assay were used to detect the effects of different doses of cisplatin on the proliferation of HepG2 cells. Flow cytometry and aging-related β-galactosidase staining were used to detect cell cycle and senescence. RT-PCR was used to detect the expression of TP53 and P21 And P19 mRNA expression levels were detected by Western blotting P53 and P21 protein expression levels. Results Cisplatin could induce irreversible growth arrest and cell cycle arrest in HepG2 cells. After 2.0μg / ml cisplatin on HepG2 aging-related β-galactosidase staining was positive and in a time-dependent manner. In the process of cisplatin-induced senescence, the expression of P19 increased, reached a peak at 48 h after induction, and then gradually decreased. The expression levels of P53 and P21 continued to increase. Conclusion This study suggests that cisplatin induces the senescent phenotype of hepatocellular carcinoma cells. This result provides a basis for further study of its anti-hepatocellular carcinoma mechanism.