一种双基因干预的转基因小鼠模型用于HBVX致肝癌分子机制的研究

The HBV X protein (HBx) is essential for virus replication in vivo and has been postulated to be associated with initiation and progression of hepatocellular carcinoma.. TSPX enhances the degradation of HBx through the ubiquitin-proteasome pathway. TSPX interacts with both HBx and a proteasome 19S lid subunit RPN3 via its C-terminal acidic tail. TSPX abrogates the RPN3- depedent stabilization of HBx, suggesting that TSPX and RPN3 act competitively in regulation of HBx stability.So, TSPX-induced HBx degradation could play key role(s) in hepatocarcinogenesis among HBV-infected HCC patients.. The different expression frequencies of TSPY transcript in WD and PD, together with the structure relationship of TSPY to SET and NAP, suggests that TSPY possibly involve in hepatocarcinogenesis and reversely involve in progression of HCC.. To establish an induced liver cell specific TSPX-RNAi/TSPY- cDNA transgenic mouse model, the transgenic mice liver tissue TSPX gene is the Knock down and the TSPY gene be the Knock in phenotype, other organizations TSPX / TSPY gene normal expression , in order to imitate hepatic precancerous lesions, the cellular microenvironment. .After the tumor suppressor genes TSPX low expression and oncogene TSPY over-expression, the phenotypic changes of liver function and sensitivity to chemical carcinogenesis agent were observed，whether the promotion of primary liver cancer phenotype after transfection HBVx, elucidate the molecular mechanism of interaction by the three, And then understanding the molecular mechanisms of malignant transformation is important.

HBVx是HBV在肝细胞中复制所必须的编码蛋白，它能激活许多与细胞粘附、增殖、生长相关的基因和信号转导途径，是引发肝细胞癌变的始动因素。肝细胞中抑癌基因TSPX可以促进HBV-x在泛素依赖的蛋白酶系统中降解，而癌基因TSPY在肝癌细胞中高表达， HBV-x、TSPX和TSPY三者关系的失衡可能是肝细胞癌变的重要机制之一。 为阐明他们之间的关系及在肝癌发病中的作用，我们建立肝细胞特异性TSPX /TSPY转基因小鼠模型，使肝组织中TSPX被 Knock down和TSPY 被Knock in，而其他组织TSPX/TSPY基因正常表达，从而分析其表型变化，观察肝细胞中TSPX低表达、TSPY过表达后，对肝功的影响，对化学诱癌剂敏感性的改变，及转染HBVx后是否能促进原发性肝癌的发生，阐明三者相互作用的分子机制，对肝细胞癌变分子机制的再认识有重要意义。

HBVx 是HBV 在肝细胞中复制所必须的编码蛋白，它能激活许多与细胞粘附、增殖、生长相关的基因和信号转导途径，是引发肝细胞癌变的始动因素。肝细胞中抑癌基因TSPX 可以促进HBV-x 在泛素依赖的蛋白酶系统中降解，而癌基因TSPY 在肝癌细胞中高表达， HBV-x、TSPX 和TSPY 三者关系的失衡可能是肝细胞癌变的重要机制之一。 为阐明他们之间的关系及在肝癌发病中的作用，我们建立肝细胞特异TSPX /TSPY 转基因小鼠模型，使肝组织中TSPX 被 Knock down 和TSPY 被Knock in，而其他组织TSPX/TSPY 基因正常表达，从而分析其表型变化，观察肝细胞中TSPX 低表达、TSPY 过表达后，对肝功的影响，对化学诱癌剂敏感性的改变，及转染HBVx 后是否能促进原发性肝癌的发生，阐明三者相互作用的分子机制，对肝细胞癌变分子机制的再认识有重要意义。