中文摘要：

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发病机制中的作用奠定了基础。方法分别将表达HBx基因的重组腺病毒Ad-HBx-GFP和空载体Ad-GFP转染小鼠肝前体细胞,再经小鼠门静脉注射后3、7、14 d取血清和肝组织,检测血清丙氨酸氨基转移酶（ALT）变化情况,观察绿色荧光蛋白表达情况,免疫组织化学法观察HBx阳性染色,并取表达高峰在第7天的肝组织,进行Western blot法和实时荧光定量PCR（Real-time PCR）法分别检测HBx的蛋白及mRNA表达。结果成功构建了表达HBx并靶向针对肝前体细胞的小鼠模型,绿色荧光蛋白表达量、免疫组化阳性细胞数量及血清ALT均在注射后第7天达到高峰,RT-PCR及Western blot均检测到第7天14-19/Ad-HBx-GFP组小鼠肝组织目的基因及蛋白的特异性表达,与14-19/Ad-GFP组及生理盐水注射组相比结果具有统计学差异。结论成功构建了表达HBx并靶向针对肝前体细胞的小鼠模型,操作可靠,重复性好,效果稳定,不仅对探讨原发性肝癌的发生机制奠定了基础,也为肿瘤动物模型的构建提供了新的思路。

英文摘要：

Objective To construct hepatic progenitor cells（HPCs） expressing hepatitis B virus X（HBx） and establish a mouse model by portal vein injection of the cells,in order to provide a platform to study the effect of HBx on HPCs and the mechanism of hepatocellular carcinoma（HCC）.Methods The HPCs（cell line 14-19） were transfected by recombinant adenovirus vectors expressing HBx gene（Ad-HBx-GFP） as well as empty vector（Ad-GFP） separately,and the transfected HPCs were infused to the mouse from the portal vein.The control group was given equivalent normal saline.The change of serum alanine aminotransferase（ALT） was detected on the 3th,7th and 14 th days.The expression of HBx and green fluorescent protein（GFP） in liver tissue was detected by immunohistochemical technique and frozen section method.The expression of HBx at mRNA and protein levels was measured by RT-PCR and Western blotting.Results The mouse model expressing HBx was successfully established.The GFP expression,serum ALT level and HBxpositive cells reached the peaks on the 7th day.Compared with the control group and the 14-19 / Ad-GFP group,the 14-19 / Ad-HBx-GFP group showed specific expression of HBx mRNA and protein in the liver tissue.Conclusion The mouse model expressing HBx is successfully established with reliable operation and good repeatability,which lays a basis for exploring the mechanism of HCC and provides a new thought for establishing a tumor animal model.