【作者】 何柳兴；

【导师】 冯涛；

【作者基本信息】 重庆医科大学 ， 生物化学与分子生物学， 2006， 硕士

【摘要】 乙型肝炎病毒(hepatitis B virus,HBV)编码的X蛋白在HBV致癌过程中发挥着关键性的作用。HBX作为反式作用因子,不能直接和双链DNA结合,而是通过蛋白与蛋白间的相互作用行使功能,可以调节转录、引起DAN修复蛋白质失活以及激活信号转导通路。Wnt信号传导途径在胚胎发育和肿瘤发生过程中起着十分重要的作用,而β-连环蛋白(β-catenin)是这个信号传导途径中的一个重要成员。HBX和β-catenin关系的研究对HBV导致肝癌的发生机制、肝癌的预防和治疗都有重要意义,为阐明Wnt信号转导途径中β-catenin在肝癌发生和发展中的作用提供理论依据。本课题采用带有GFP的腺病毒高表达载体系统有效转染人正常肝细胞系L02,并用Western blot证实HBX是否能有效在L02细胞中表达。用RT-PCR、免疫细胞化学方法分析HBx对正常细胞中β-catenin定位及基因表达的变化,并通过免疫共沉淀的方法来分析HBX与β-catenin、HBX与APC蛋白之间是否存在直接相互作用,了解HBX对正常肝细胞中β-catenin的影响及作用机制,为进一步研究HBX导致肝癌的机制及其预防治疗奠定基础。更多还原

【Abstract】 HBx plays a critical role in the HBV-associated hepatoma. As a transcriptional activator, HBx functions through interaction between protein and protein ,instead of binding directly with DNA. HBx can regulate cell growth, cellular signal transduction.β-catenin is an essential component of Wnt signaling pathway which plays an important role in the development of HCC. So the investigation of relationship between HBx andβ-catenin will contribute to explain the mechanism of development of HCC,to provide evidence for the important role ofβ-catenin in Wnt signaling pathway and to have better prevetion and therapy of HCC.In this study, recombinant adenovirus vector that expressed HBx gene with GFP was used to effectively transfect normal human hepatocyte cell line L02.Then, the changes of mRNA expression and location ofβ-catenin were observed by RT-PCR and immunocytochemistry. we used the method of co-immunoprecipitation to find out whether there exists interaction between HBX andβ-catenin、HBX and APC. It is helpful to pave a way to further study function of HBX and to look for a new cure更多还原