奥沙利铂对肝癌细胞HuH-7细胞周期的影响

Effect of Oxaliplatin on Cell Cycle of Hepatocellular Carcinoma Cell Line HuH-7

目的探讨奥沙利铂(Oxaliplatin,L-OHP)对人肝癌细胞株HuH-7细胞周期的影响及其相关机制,为其应用于原发性肝细胞癌临床治疗提供理论依据。方法 MTT方法检测L-OHP对肝癌细胞HuH-7生长的抑制作用;流式细胞术(FCM)检测L-OHP诱导HuH-7细胞周期阻滞的作用;RT-PCR和Western blot方法观察L-OHP对细胞周期调节因子Cyclin D1、CDK2、CDK4及p16、p21、p53表达的影响。结果 MTT结果显示L-OHP对HuH-7细胞增殖抑制率随药物浓度增加及作用时间延长有升高趋势,呈时间剂量依赖性。L-OHP能够诱导HuH-7细胞周期阻滞于S期,并可下调CDK4和Cyclin D1蛋白的表达,上调p21蛋白的表达,低浓度上调p53表达而高浓度下调p53表达,CDK2和p16表达无明显变化。结论 L-OHP可能通过影响CDK4、Cyclin D1及p21的活性使HuH-7细胞阻滞在S期,从而抑制肝癌细胞HuH-7的增殖。

Objective To study the effect of Oxaliplatin（L-OHP）on cell cycle of hepatocellular carcinoma（HCC）cell line HuH-7 and the involved mechanism in order to provide the theoretical foundation for the chemotherapy of HCC.Methods Inhibitory effect of L-OHP on the proliferation of HuH-7 cells was determined by MTT assay.Cell cycle distribution was shown by flow cytometry.Cyclin D1,CDK2,CDK4,p16,and p21 expression levels were detected by uisng RT-PCR and Western blot.Results MTT method revealed that L-OHP could inhibit the proliferation of HCC HuH-7 cells in a dose-and time-dependent manner.L-OHP could cause S cell cycle arrest in HuH-7 cells,down-regulate the levels of CDK4 and Cyclin D1,and up-regulate the level of p21.L-OHP could up-regulate the level of p53 in low concentration and down-regulate the level of p53 in high concentration.Meanwhile,there were no noticeable changes in CDK2 and p16.Conclusion L-OHP can inhibit the proliferation of HuH-7 cells by inducing cell blockage at S stage,which may be resulted from the activity of CDK4,Cyclin D1 and p21.