组蛋白去乙酰化酶抑制剂与5-FU合用对肝癌细胞HepG2的影响

Effect of combined application of HDACi and 5-fluorouracil on hepatoma HepG2 cells

目的:探讨MS-275和5-FU联用对HepG2细胞周期和凋亡的影响,并对其机制进行初步探讨。方法:将细胞分成对照组、MS-275组、5-FU组和联合用药组。流式细胞仪检测各组细胞凋亡和周期变化;Westernblot分析各组Bcl-2、Bax、CyclinD1、P21蛋白表达。结果:MS-275和5-FU联用能抑制肝癌细胞生长,诱导G0-G1期阻滞,促进细胞凋亡。上述效应具有时间和剂量依赖性。二者联用可以使P21蛋白上调,Bcl-2、CyclinD1蛋白下调,Bax蛋白无明显变化。结论:5-FU和MS-275联用能提高HepG2细胞凋亡率和周期阻滞,其机制与下调CyclinD、Bcl-2蛋白的表达、上调P21蛋白表达有关。

Objective：To investigate the influence of combined application of histone deacetylase inhibitor（HDACi） MS-275 and 5-FU on the apoptosis and cycle arrest on human hepatoma cell line HepG2 and unclose the related mechanism. Methods ： Divided all the cells into 4 groups：control group,MS-275 group,5-FU group and drug combination group. Flow cytometry （FCM） was used＇to examine the effects of 5-FU w/th MS-275 on the apoptosis and cell cycle of HepG2 cells. Bcl-2, Bax, CyclinD1 and P21 protein were determined by Western blot assay. Results：5-FU and MS-275 combination could inhibit HepG2 cell growth through G0-G1 arrest,and induce apoptosis,both time and dose dependently. The combination of two agents increased P21 protein levels and decreased Bcl-2, Cy- clinD1 protein levels. The levels of Bax protein were not changed. Conclusion：The combination of 5-FU and MS-275 can induce apop- tosis and cell cycle arrest,the effect might be associated with down-regulating expression of Cyclin D1 and Bcl-2 protein and upregulating the expression of P21 protein.