表达hbx的肝前体细胞肝内移植小鼠模型的构建 - 第三军医大学学报.doc

表达HBx的肝前体细胞经肝内移植小鼠模型的构建 钟沁1,2，张超1,2,何雪梅1，2，胡代曦12，张锡峰12,黄佳祎1，3，冯涛1,2* 1.重庆医科大学分子医学与肿瘤研究中心，重庆，400016 2.重庆医科大学基础医学院生物化学与分子生物研究中心，重庆，400016 3.重庆医科大学基础医学院病理生理学教研室，重庆，400016 [摘要] 目的：构建表达乙肝病毒X基因（HBx）的肝前体细胞，并经小鼠门静脉注射建立肝内移植模型，为进一步研究HBx 对肝前体细胞的作用和影响及其在原发性肝癌（HCC）Establishment of mouse models via Intrahepatic transplantation of Progenitor Cells Expressing Hepatitis B Virus X Zhong Qin1,2,Zhang Chao1,2,He Xuemei1,2，Hu Daixi12,Zhang Xifeng1,2,Huang Jiayi1,3,Feng Tao1,2 Research Center of Molecular Medicine and Cancer， Chongqing 400016，China; Department of Pathophysiology, College of Basic Medicine, Chongqing 400016，China; Department Pathophy Siology College of Basic Medicine, Chongqing 400016, China Abstract Objective:To eastablish Hepatic progenitor cells(HPCs) expessiong Hepatitis B virus X(HBx）and develop a mouse modles Via the portal vein injection of the cells.It can provided a platform to study the effect of HBx on HPCs and the mechanisms of Hepatocellular carcinoma(HCC). Methods:recombinant adenovirus were transfected into 14-19 cells;mice were received a injection of the cells via portal vein.The expression of HBx and GFP in liver tissues were detected by immunohistochemistry technique and frozen section .The expression levels of HBx mRNA and HBx prteins were measured by PCR and Western blotting.Results:Compared with the controls,the expression of HBx in liver section or at mRNA and protein levels was positive. Conclusion:The established mice model allows the HBx aimed at HPCs directly,it has good?stability?and?maneuverability, and it help to dissect the effect of HBx on HPCs. Key words HBx; Hepatic progenitor cells;green fluorescent;proteinPortal vein;Mouse model 原发性肝癌（Hepatocellular carcinoma,HCC）,通过多种复杂的途径与HBV相关性肝癌的发生、发展有着密切的联系[2-3]，但其具体机制还不是很清楚。肝前体细胞（hepatic progenitor cells ，HPCs),是一类存在于肝组织内具有干细胞特性的细胞，在肝损伤过程中及特定环境因素下常伴有肝前体细胞的增殖，其甚至能向肝细胞癌方向发展,近年来研究显示肝干细胞可能是肝癌发生的起始细胞，其可能与肝干细胞分化受阻有关[4-6]。本课题组前期相关研究成果显示，HBx能抑制小鼠胚胎干细胞的凋亡，通过调控信号通路影响肝前体细胞的增殖、分化，并能诱导肝前体细胞上皮-间质转化（Epitheli