乙肝病毒X基因转化人肝细胞裸鼠成瘤实验及其机制  

Study on tumor formation of HBV X gene-transformed hepatocyte in nude mice

作　　者：郑芳 刘国珍[2] 

机构地区：[1]长沙市第一医院感染科,长沙410005 [2]中南大学湘雅医院感染科

出　　处：《中国医师杂志》2013年第5期590-594,共5页Journal of Chinese Physician

基　　金：湖南省科技厅科技计划项目（2007SK3033）

摘　　要：目的在裸鼠体内证实乙肝病毒X基因（HBx）能否转化人肝细胞形成移植瘤；并研究裸鼠成瘤的可能机制。方法利用已成功构建的高表达HBx基因的pCMVX／QSG7701细胞株作为实验组、以表达空白质粒的pRcCMV2／QSG7701细胞株和QSG7701细胞株为对照组接种于裸鼠皮下，观察裸鼠能否成瘤。HE染色、显微镜检研究移植瘤的组织形态学特征。成瘤率的比较采用Fisher＇s精确检验。免疫组化方法研究的pCMYX／QSG77细胞、pRcCMV2／QSG7701细胞和QSG7701细胞的p53蛋白、C-Myc蛋白的表达情况。结果接种pCMVX／QSG7701细胞株的裸鼠在第2周开始出现移植瘤生长，周围器官和组织未发现转移灶。对照组至接种后第35天无移植瘤生长。HE染色证实移植瘤为肝细胞癌组织。pCMVX／QSG77细胞、pRcCMV2／QSG7701细胞和QSG7701细胞中均有突变型p53蛋白及c-Myc蛋白的表达。pCMVX／QSG7701细胞中突变型p53蛋白及C—Myc蛋白的表达量较pRcCMV2／QSG7701细胞、QSG7701细胞明显增高（P〈0．01）。结论HBVX基因表达可以直接导致肝细胞癌变。在蛋白水平证实HBx能上调pMVX／QSG7701细胞中p53和C—Myc的表达，并可能通过这一机制导致肝细胞癌变。Objective To investigate whether HBx alone is sufficient to directly transform the non- transformed immortalized human liver cell line QSG7701 and induce hepatocellular carcinoma in vivo, and to explore preliminarily the pathogenic mechanism of transplantation tumor in nude mice. Methods The pCMVX/QSG7701 cells were vaccinated into subcutaneous tissue of nude mice. The pRcCMV2/QSG7701 and QSG7701 Cells were used as control. At the fifth weeks after vaccination, the nude mice were killed to observe whether a tumor was formed. The activity state and food intake of the nude mice was recorded. The texture, volume, and metastasis of transplantation tumor were observed grossly. The transplantation tumor was observed microscopically on the hematoxylin and eosin （HE） staining section. Immunohistochemical surfactant protein （SP） method was used to analyze the protein expression of mutant p53 and C-Myc genes. Results The transplantation tumor was occurred in all of the six nude mice vaccinated with pCMVX/ QSG7701 cells at the second week after vaccination. No metastatic tumor was found in other organs. Trans- planted tumor was not formed in all of the negative control groups. HE staining analysis confirmed that the character of transplanted tumor was hepatocellular carcinoma, p53 and C-Myc proteins were expressed in pCMVX/QSG7701, pRcCMV2/QSG7701, and QSG7701 cells, and their expression levels in the pCMVX/ QSG7701 cells were significantly higher than those in the pRcCMV2/QSG7701 and QSG7701 cells, respec- tively（ P 〈0. 01 ）. Conclusions HBx alone is sufficient to directly transform the non-transformed immor- talized human liver cell line QSG7701 and induce hepatocellular carcinoma in vivo through up-regulating the expression of mutant p53 and C-Myc genes.

关 键 词：肝炎病毒 乙型 遗传学 肝细胞 癌 肝细胞 肝肿瘤 实验性 

分 类 号：R735.7[医药卫生—肿瘤]