苦参素对肝癌耐药细胞株HepG2/ADM的作用及机制研究

Effect and Mechanism of Oxymatrine on Inhibiting ERK/MAPK Axis on Hepatocellular Carcinoma Resistant Cell Line HepG2/ADM

目的探讨苦参素对肝癌耐药HepG2/ADM细胞的影响及其具体机制。方法不同浓度苦参素作用于HepG2/ADM细胞,MTT法检测细胞存活率;不同浓度阿霉素处理HepG2细胞和HepG2/ADM细胞计算耐药指数,苦参素联合不同浓度阿霉素处理HepG2/ADM细胞,计算苦参素对HepG2/ADM细胞耐药性逆转倍数;HepG2/ADM细胞分为对照组、苦参素组、阿霉素组和苦参素+阿霉素组,Transwell实验检测细胞迁移和侵袭能力,流式细胞仪检测细胞凋亡率,蛋白印迹法检测p-ERK1/2、P糖蛋白(P-gp)、肺耐药相关蛋白(LRP)和乳腺癌耐药蛋白(BCRP)相对表达量。结果细胞存活率随苦参素浓度升高逐渐降低(P<0.05);HepG2/ADM细胞的耐药指数为4.77,苦参素耐药性逆转倍数为2.57。与对照组比较,苦参素组和阿霉素组细胞迁移和侵袭能力以及p-ERK1/2、P-gp、LRP和BCRP蛋白相对表达量降低,细胞凋亡率升高(P<0.05);与苦参素组和阿霉素组比较,苦参素+阿霉素组细胞迁移和侵袭能力以及p-ERK1/2、P-gp、LRP和BCRP蛋白相对表达量降低,细胞凋亡率升高(P<0.05)。结论苦参素对肝癌耐药HepG2/ADM细胞的耐药性具有一定逆转作用,而且可降低耐药细胞迁移和侵袭能力,诱导细胞凋亡,其可能是通过抑制ERK/MAPK信号通路发挥作用。

Objective To investigate the effect of oxymatrine on hepatocellular carcinoma resistant HepG2/ADM cells and its specific mechanism.Methods HepG2/ADM cells were treated with different concentrations of oxymatrine,and the cell survival rate was detected by MTT assay.HepG2 cells and HepG2/ADM cells were treated with adriamycin at different concentrations to calculate drug resistance index.HepG2/ADM cells were treated with oxymatrine at different concentrations to calculate drug resistance reversal ratio of matrine to HepG2/ADM cells.HepG2/ADM cells were divided into the control group,oxymatrine group,adriamycin group and oxymatrine+adriamycin group.Cell migration and invasion ability were detected by Transwell assay,cell apoptosis rate was detected by flow cytometry.Western blot was used to detect the relative expression levels of p-ERK1/2,P-glycoprotein(P-GP),lung drug resistance associated protein(LRP)and breast cancer drug resistance protein(BCRP)proteins.Results The cell survival rate decreased with the increase of oxymatrine concentration(P<0.05).The drug resistance index of HepG2/ADM cells was 4.77,and the reversal ratio of oxymatrine resistance was 2.57.Compared with the control group,cell migration and invasion ability,relative expression levels of p-ERK1/2,P-GP,LRP and BCRP were decreased in oxymatrine and adriamycin groups,and cell apoptosis rate was increased(P<0.05).Compared with oxymatrine group and adriamycin group,the cell migration and invasion ability and the relative expression levels of p-ERK1/2,P-GP,LRP and BCRP in oxymatrine+adriamycin group were decreased,and the cell apoptosis rate was increased(P<0.05).Conclusion Oxymatrine can reverse the drug resistance of HepG2/ADM cells,reduce the migration and invasion ability of cells,and induce apoptosis,possibly by inhibiting ERK/MAPK signaling pathway.