α-干扰素联合索拉菲尼对肝癌细胞增殖的抑制作用及其对STAT3通路的影响

Inhibitory effect ofα-interferon combined with sorafenib on proliferation of hepatocellular carcinoma cells and its effect on STAT3 pathway

目的:探讨α-干扰素(IFN-α)联合索拉菲尼对肝癌细胞增殖的抑制作用及其对转录激活因子3(STAT3)的影响。方法:选取人肝癌细胞株HepG2,分为空白对照组(未经任何处理的HL-60细胞)、IFN-α组(加入4000 U/ml IFN-α)、索拉菲尼组(加入2μmol/L索拉菲尼)、IFN-α+索拉菲尼组(加入4000 U/ml和2μmol/L索拉菲尼),取经药物处理后的各组细胞混悬液,继续培养24、48、72 h,采用CCK8检测HepG2细胞存活率,MTT法检测HepG2细胞增殖抑制率,行蛋白质免疫印迹(Western blot)、实时定量PCR(qRT-PCR)测HepG2细胞中STAT3、Survivin mRNA和蛋白表达,并进行细胞形态学观察。结果:空白对照组细胞形态完整,轮廓清晰,呈梭形,贴壁状态良好,IFN-α组、索拉菲尼组部分胞浆肿胀、胞膜破裂、细胞形态、胞膜界限模糊不清,IFN-α+索拉菲尼组,细胞核缩小、染色质固缩、部分细胞碎裂、死亡。IFN-α+索拉菲尼组细胞增殖抑制率高于IFN-α组、索拉菲尼组(均P<0.05)。与空白对照组、IFN-α组、索拉菲尼组比较,IFN-α+索拉菲尼组细胞G 1期细胞器增多,S、G 2期细胞减少(均P<0.05)。与空白对照组比较,IFN-α组、索拉菲尼组、IFN-α+索拉菲尼组细胞中STAT3、Survivin mRNA和蛋白表达均降低,但IFN-α组和索拉菲尼组比较无统计学差异(均P<0.05),IFN-α+索拉菲尼组低于其余三组(均P<0.05)。结论:α-干扰素联合索拉菲尼可抑制HepG2细胞增殖并诱导凋亡,其机制可能是通过调节STAT3通路发挥作用。

Objective:To investigate the inhibitory effect ofα-interferon(IFN-α)combined with sorafenib on the proliferation of hepatocellular carcinoma cell and its effect on transcriptional activator 3(STAT3).Methods:Human hepatocellular carcinoma cell line HepG2 was selected and divided into blank control group(HL-60 cells without any treatment),IFN-αgroup(adding 4000 U/ml IFN-α),sorafenib group(adding 2μmol/L sorafenib),IFN-α+sorafenib group(adding 4000 U/ml and 2μmol/L sorafenib).Cell suspension of each group after drug treatment was taken and cultured for 24,48 and 72 hours.The survival rate of HepG2 cells was detected by CCK8 and the proliferation inhibition rate of HepG2 cells was detected by MTT method.The expressions of STAT3 and Survivin mRNA and protein in HepG2 cells were measured by Western Blot and qRT-PCR,and the cell morphology was observed.Results:In the blank control group,the cells were in a complete shape,with clear outline,find-shaped,and well adherent state;in the IFN-αgroup and sorafenib group,part of the cytoplasm was swollen,the cell membrane was broken,and the cell morphology and membrane boundaries were blurred;in the IFN-α+sorafenib group,the nucleus was reduced,chromatin was contracted,and some cells were broken and died.The cell proliferation inhibition rate in IFN-α+sorafenib group was higher than that in IFN-αand sorafenib groups(all P<0.05).Compared with blank control group,IFN-αgroup and sorafenib group,cell organelles of IFN-α+sorafenib group increased in G 1 phase,and cells of S and G 2 phase decreased(all P<0.05).Compared with blank control group,STAT3,Survivin mRNA and protein expressions in IFN-αgroup,sorafenib group and IFN-α+sorafenib group were decreased(all P<0.05),but there was no significant difference between IFN-αgroup and sorafenib group(all P>0.05),and IFN-α+sorafenib group was lower than those of the other three groups(all P<0.05).Conclusion:α-interferon combined with sorafenib can inhibit proliferation and induce apoptosis of HepG2 cells,possibly by regulating STAT3 pathway.