HBx通过HIPPO信号通路导致小鼠肝癌的发生机制  

Hepatitis B virus X promotes tumorigenesis of hepatocellular carcinoma through HIPPO signaling pathway in mice

作　　者：张思遥 黄欣 王薛 杜彬 毋楠 周梦瑶 冯涛[1] ZHANG Siyao;HUANG Xin;WANG Xue;DU Bin;WU Nan;ZHOU Mengyao;FENG Tao(Molecular Medicine and Cancer Research Center,Department of Biochemistry and Molecular Biology,College of Basic Medical Sciences,Chongqing Medical University,Chongqing,400016,Chin)

机构地区：[1]重庆医科大学分子医学与肿瘤研究中心,基础医学院生物化学与分子生物研究中心,重庆400016

出　　处：《第三军医大学学报》2020年第7期664-670,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81071770)。

摘　　要：目的观察HIPPO信号通路在小鼠感染乙型肝炎病毒X(hepatitis B virus X,HBx)不同时间的变化,初步探讨HBx导致肝癌发生的机制。方法将转染HBx的肝前体细胞通过门静脉注射入小鼠体内,构建能稳定表达HBx的动物模型,分别于30、90、180、360 d取肝脏组织,HE染色观察肝脏组织病理特征,RT-PCR、Western blot以及免疫组化法检测肝脏组织中HBx的表达,RT-PCR检测HIPPO信号通路相关因子MST1、YAP1 mRNA的表达,Western blot检测MST1、YAP1、p-MST1、p-YAP1蛋白的表达。结果成功构建能在小鼠肝脏中稳定表达HBx的动物模型并发展为肝癌,Western blot结果显示,与对照组相比,HBx组小鼠肝脏组织在术后30 d即出现MST1表达下调,p-MST1表达上调(P<0.01);YAP1表达上调,p-YAP1表达下调(P<0.05),RT-PCR与Western blot结果一致。这种改变一直持续至肿瘤发生。结论HBx可以导致小鼠肝癌发生,其机制可能与长期影响HIPPO信号通路的表达有关。Objective To explore the changes in HIPPO signaling pathway in mouse liver tissues expressing hepatitis B virus X protein(HBx)and explore the mechanism of HBx for causing hepatocellular carcinoma(HCC).Methods Immortalized murine liver progenitor cells transfected with HBx gene were injected in mice via the hepatic portal vein.At 30,90,180,and 360 d after the cell injection,the liver tissues of the mice were sampled to examine HBx expression using real-time PCR,Western blotting and immunohistochemistry;the expression levels of the key factors in the HIPPO signaling pathway(MST1,YAP1,p-MST1,and p-YAP1)were detected using real-time PCR and Western blotting.Results We successfully established a mouse model with HBx expression in the liver tissues.The results of both real-time PCR and Western blotting showed that the expression levels of MST1 and p-YAP1 were down-regulated and YAP1 and p-MST1 were up-regulated in the liver tissues of the mouse model,and such changes persisted till the occurrence of HCC.Conclusion HBx causes HCC in mice possibly by persistently affecting HIPPO signaling pathway.

关 键 词：HBX 动物模型 肝癌 HIPPO信号通路 

分 类 号：R372.1[医药卫生—病原生物学] R730.23[医药卫生—基础医学]