转染HBx改变Notch信号诱导肝癌HepG2细胞株多药耐药性研究  

A Study on Multidrug Resistance of Cell Line HepG2 induced by Changing Notch Signaling Pathway through HBx Transfection in Hepatocellular Carcinoma

作　　者：徐桐红 于冬艳 任玲 XU Tonghong;YU Dongyan;REN Ling(The Seventh Department of Internal Medicine,Tumour Hospital of Liaoning Province　Tumour Hospital of China Medical College,Shenyang 110042,China)

机构地区：[1]中国医科大学肿瘤医院辽宁省肿瘤医院内七科,沈阳110042

出　　处：《解放军医药杂志》2018年第1期38-41,共4页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：辽宁省自然科学基金项目(2014020101)

摘　　要：目的研究转染HBx对Notch信号通路的影响,分析其诱导肝癌多药耐药(MDR)的发生机制。方法选用人肝癌HepG2细胞株进行转染,根据HBx转染情况将细胞分为转染HBx细胞组(HepG2-HBx组)、转染空载体细胞组(HepG2-con组)和空白细胞组(HepG2组)。实时定量聚合酶链反应(PCR)检测3组细胞HBx mRNA表达。蛋白免疫印迹法(Western-blot)检测3组细胞HBx、Notch-1和多药耐药蛋白(MRP)表达,CCK-8法检测3组细胞增殖活性和耐药性。结果 HepG2-HBx组有HBx片段和HBx蛋白表达,HepG2-con组和HepG2组未见HBx mRNA和蛋白表达。与HepG2组和HepG2-con组比较,HepG2-HBx组细胞增殖加快,Notch-1和MRP蛋白表达增加(P<0.05)。细胞周期检测结果显示,与HepG2组和HepG2-con组比较,HepG2-HBx组G0/G1期细胞显著减少,S期细胞增加(P<0.05,P<0.01)。MDR实验显示,HepG2-HBx组对5种药物的耐药指数增加明显(P<0.05)。结论转染HBx的肝癌HepG2细胞株可能通过诱导Notch-1与MRP的过表达,最终导致肝癌HepG2细胞增殖力增强和MDR的发生。Objective To investigate incidence mechanism of multidrug resistance induced by affecting Notch signaling pathway through HBx transfection in hepatocellular carcinoma(HCC).Methods Human hepatoma HepG2 cell line was transfected,and the samples were divided into transfected HBx cell group(HepG2-HBx group),transfected cell group with empty vector(HepG2-con group)and blank cells group(HepG2 group).In 3 groups,HBx mRNA expressions were detected by quantitative polymerase chain reaction(PCR),and HBx,Notch-1 and multidrug resistance-associated proteins(MRP)expressions were detected by Western-blot method,and cell conditions of proliferation activity and multidrug resistance were detected by CCK-8.Results HBx fragments and HBx expressions were detected in HepG2-HBx group,while no HBx mRNA and protein expressions could be found in HepG2-con and HepG2 groups.Compared with those in HepG2-con and HepG2 groups,cell proliferation activity was significantly enhanced,and Notch-1 and multidrug resistance-associated proteins(MRP)protein expressions were significantly increased in HepG2-HBx group(P<0.05).Cell cycle detection showed that in HepG2-HBx group,cell number in G0/G1 phase was significantly decreased,while cell number in S phase was significantly increased compared with those in HepG2-con and HepG2 groups(P<0.05,P<0.01).Multidrug resistance(MDR)examination showed that resistance indexes for 5 drugs were significantly increased in HepG2-HBx group(P<0.05).Conclusion HepG2 cell line in hepatocellular carcinoma transfected by HBx can finally lead to incidence of increasing HepG2 proliferation and multidrug resistance through inducing Notch-1 and MRP overexpressions.

关 键 词：肝肿瘤 HEPG2细胞 HBX 转染 Notch信号通路 多药耐药相关蛋白质类 

分 类 号：R735.7[医药卫生—肿瘤]