索拉菲尼抑制肿瘤细胞PKM2蛋白表达

Experimental research about sorafenib inhibiting PKM2 protein expression in cancer cell line

目的:探讨索拉菲尼对肿瘤细胞中PKM2蛋白表达的影响。方法:以人肝癌细胞HepG2、人胆管癌细胞QBC939、人肺癌细胞NCI-H446、人宫颈癌细胞Hela为研究对象,体外培养上述肿瘤细胞;光镜下观察不同肿瘤细胞经不同浓度索拉菲尼处理后细胞形态学改变;CCK-8法检测索拉菲尼对上述肿瘤细胞增殖的影响。用Western blot检测索拉菲尼对上述细胞的PKM2表达变化。结果:光镜下观察可见随着索拉菲尼浓度的增加,肿瘤细胞离巢、凋亡,并且随着浓度的增加而增加。细胞增殖实验可见,索拉菲尼显著抑制HepG2、QBC939、NCI-H446、Hela细胞的增殖,不同浓度(2.5μmol/L、5.0μmol/L、10.0μmol/L、20.0μmol/L)的索拉菲尼处理上述细胞48 h后,各药物处理组与对照组比较(P均<0.01),呈剂量效应关系;索拉菲尼处理上述肿瘤细胞48 h的IC50分别为10.61μmol/L、13.88μmol/L、15.66μmol/L、12.86μmol/L。Western blot检测结果显示,不同浓度的索拉非尼显著抑制HepG2、QBC939、NCI-H446、Hela细胞的PKM2的蛋白表达,并且表达与浓度呈剂量关系。蛋白表达半定量分析可见,各肿瘤细胞经过索拉菲尼处理后与对照组比较,蛋白表达差异有统计学意义(P<0.05)。结论:索拉菲尼能抑制人多种肿瘤细胞增殖并显著抑制PKM2蛋白表达。

Objective:To explore the effect of sorafenib on PKM2 protein expression in tumor cells.Methods:Human hepatic tumor cell line HepG2,human cholangiocarcinoma cell line QBC939,human lung cancer cell line NCI-H446 and human cervical cancer cell line Hela were cultured in vitro.The morphological changes of cell apoptosis induced by different concentrations of sorafenib were observed under optical microscope.CCK-8 array was used to detect the influence of sorafenib on tumor cell proliferation.Western blot was used to detect the influence of sorafenib on PKM2 expression.Results:Under optical microscope,apparent anoikis and apoptosis occurred in tumor cells and increased with the increase of sorafenib concentration.In the proliferation experiment,sorafenib significantly inhibited the proliferation of HepG2,QBC939,NCI-H446 and Hela tumor cells in a dose dependent manner.Different concentrations of sorafenib(2.5 μmol/L,5.0 μmol/L,10.0 μmol/L,20.0 μmol/L) treated the tumor cells for 48 h and were compared with the control group(P all<0.01).IC50 of sorafenib after 48 h treatment was 10.61 μmol/L,13.88 μmol/L,15.66 μmol/L,12.86 μmol/L,respectively.Western blot results showed that different concentrations of sorafenib significantly inhibited the expression of PKM2 in HepG2,QBC939,NCI-H446 and Hela cells in a dose dependent manner.Semi-quantitative analysis of protein expression showed that the PKM2 protein expression after different concentrations of sorafenib in treating tumor cells was significantly different from the control group(P<0.05).Conclusion:Sorafenib can effectively inhibit the proliferation of a variety of human tumor cells and PKM2 expression.