摘要
目的探讨HBx上调TGF-β/TβR-Ⅱ通路对肝细胞癌索拉菲尼耐药的影响。方法选择稳定转染HBX蛋白的HepG2-HBx细胞与转染pcDNA3.1质粒的HepG2-3.1细胞,采用索拉菲尼进行药物处理,运用MTT法来检测索拉菲尼对细胞生长的抑制率,接着流式实验对细胞凋亡率加以检测。采用免疫印迹方法检测TGF-β/TβR-Ⅱ通路中TβR-Ⅱ的蛋白水平。结果 HepG2-HBx细胞的HBX蛋白表达明显高于普通的HepG2-3.1细胞(P<0.05)。MTT检测结果显示索拉菲尼对HepG2-HBx细胞的IC_(50)值为HepG2-3.1细胞的4.65倍。流式检测结果显示索拉菲尼作用24 h后,HepG2-HBx细胞的凋亡率要明显低于HepG2-3.1细胞的凋亡率(P<0.05)。免疫印迹检测显示HepG2-HBx细胞较HepG2-3.1细胞的多药耐药蛋白TGF-β表达量明显增高(P<0.05)。结论 HBx可通过上调TGF-β/TβR-Ⅱ通路的表达,从而提高索拉菲尼对肝癌细胞的耐药性,降低细胞凋亡率,HBx是引起肝癌细胞耐药的重要因素之一。
Objective To investigate the effect of activation of TGF-β/TβR-Ⅱ pathway by HBx onsorafenib resistance in hepatocellular carcinoma. Methods HBX protein was stably transfected into HepG2-HBxcells and pcDNA3.1 plasmid was transfected into HepG2-3.1 cells,followed with the sorafenib drug treatment. Cellgrowth inhibition rate and apoptosis were detected by MTT assay and the flow cytometry,respectively. Western blotwas used to detect the protein expression of TβR-Ⅱ. Results HBX protein expression in HepG2-HBx cells wassignificantly higher than that in the normal HepG2-3.1 cells(P〈0.05). The MTT test showed that the IC(50) value ofsorafenib on HepG2-HBx cells was 4.65 times than that of HepG2-3.1 cells. Flow cytometry showed that the apoptoticrate of HepG2-HBx cells was significantly lower than that of HepG2-3.1 cells after 24 h sorafenib treatment(P〈0.05). Western blot showed that the expression of multidrug resistance protein TβR-Ⅱ in HepG2-HBx cells wassignificantly higher than that in HepG2-3.1 cells(P〈0.05). Conclusions HBx can activate the expression ofTGF-β/TβR-Ⅱ pathway,so as to improve the drug resistance of sorafenib on hepatocellular carcinoma cells and reducethe apoptosis rate. HBx is one of the important factors leading to drug resistance of hepatocellular carcinoma cells.
作者
邢健鹦
何远学
曾洪兰
陈家诚
周钰
沈群
周文珂
陈晓玲
叶文玉
武金才
XING Jianying;HE Yuanxue;ZENG Honglan;CHEN Jiacheng;ZHOU Yu;SHEN Qun;ZHOU Wenke;CHEN Xiaoling;YE Wenyu;WU Jincai(Appointment Service Center,Hainan General Hospital,Haikou 570311,China)
出处
《实用医学杂志》
CAS
北大核心
2018年第11期1762-1765,共4页
The Journal of Practical Medicine
基金
国家自然科学基金项目(编号:81660489)