HEPATIC CARCINOGENESIS INDUCED BY HEPATITIS B VIRUS PreS2 MUTANT

长期目标：丙型肝炎病毒（HCV）是引起肝细胞性肝癌的最重要原因之一， carcinoma.这项提案的长期目标是通过以下方式了解肝癌发生的机制： 丙型肝炎病毒，特别关注病毒核心蛋白的作用。 基本原理：我们先前在HCV感染的Raji细胞和HCV核心蛋白转基因小鼠上的发现， 提出了HCV诱导的肝癌发生的几种途径，包括 炎症和HCV蛋白对DMAdamage修复的抑制。此外，激活 Toll样受体4（TLR 4）通过HCVNS 5A蛋白表达，提示HCV和 酒精/糖尿病，因为后者诱导对内毒素的敏感性增强。这些都是新的机制， 由细胞质RNA病毒引起的病毒肿瘤发生。具体目标：在这个子项目中，我们将使用两种培养细胞 HCV核心蛋白转基因小鼠，以探讨HCV感染的可能机制。 HCV肿瘤发生。提出了三个具体目标：1）建立炎症在HCV中的重要性 肿瘤发生HCV感染对NFkB和IKK β在HCV感染中的影响将在体外培养中研究。 细胞系和IKK β条件性敲除小鼠中。此外，表达HCV蛋白的转基因小鼠 将产生B细胞中的细胞因子，以检查炎性细胞因子对肿瘤发生的可能影响。（二） 使用具有增强的TLR 4表达的NS 5A转基因小鼠，我们将研究可能的协同作用。 HCV和酒精/糖尿病之间的联系将产生Core/NS 5A转基因-TLR 4-/-小鼠，并且肿瘤细胞将被转染。 将酒精/肥胖诱导的发病率与核心/NS 5A Tg小鼠进行比较，以评估 TLR4。3)探讨HCV抑制DNA损伤修复的机制。DMAdamage传感器和 将在HCV感染的细胞或表达HCV核心蛋白的细胞中研究应答复合物。各种 DMA修复途径将被研究。最后，HCV感染和核心蛋白或NS 3蛋白的蛋白质组学分析， 表达细胞中的蛋白质，以鉴定HCV的其他潜在靶蛋白。这些研究 预期将产生关于HCV肿瘤发生的可能机制的知识。

Long-term objectives: Hepatitis C virus (HCV) is one of the most important causes of hepatocellular carcinoma. The long-term goal of this proposal is to understand the mechanism of hepatocarcinogenesis by hepatitis C virus, specifically focusing on the role of the viral core protein. Rationale: Our previous findings on HCV-infected Raji cells and HCV core protein-transgenic mice have suggested several pathways for HCV-induced hepatocarcinogenesis, including the possible role of inflammation and of the inhibition of DMAdamage repair by HCV proteins. Furthermore, the activation of toll-like receptor 4 (TLR4) by HCV NS5A protein suggested possible synergism between HCV and alcohol/diabetes since the latter induces enhanced sensitivity to endotoxin. These are novel mechanisms of viral oncogenesis by a cytoplasmic RNA virus. Specific aims: In this subproject, we will use both culture cell lines infected by HCV in vitro and HCV core protein-transgenic mice to explore the potential mechanisms of HCV oncogenesis. Three specific aims are proposed: 1) Establish the importance of inflammation in HCV oncogenesis. The effects of HCV infection on NFkB and IKKbeta in HCV infection will be studied in cultured cell lines and in IKKbeta-conditional knockout mice. Furthermore, transgenic mice expressing HCV proteins in B cells will be generated to examine the possible effect of inflammatory cytokines on oncogenesis. 2) Using NS5A-transgenic mice, which have enhanced TLR4 expression, we will study the possible synergism between HCV and alcohol/diabetes. Core/NS5A transgenic-TLR4-/- mice will be generated, and the tumor incidence induced by alcohol/obesity will be compared with the core/NS5A Tg mice to assess the role of TLR4. 3) Study the mechanism of inhibition of DMAdamage-repair by HCV. The DMAdamage sensor and response complexes will be studied in HCV-infected cells or cells expressing HCV core protein. Various DMA repair pathwayswill be studied. Finally proteomic analysis in HCV-infected and core- or NS3- expressing cells will be performed to identify other potential target proteins of HCV. These studies are expected to generate knowledge on the possible mechanism of HCV oncogenesis.