preS1调控GP73/NF-κB信号通路导致HBV相关性肝癌发生发展的作用机制研究

Chronic HBV infection is very easy to develop into hepatocellular carcinoma, which seriously threatens the health of 120 million chronic carriers of HBV in China. At present, the molecular mechanism of HBV-related hepatocellular carcinogenesis is still lacking in academia. Our previous research found that: preS1 encoded by HBV can up-regulate the expression of GP73 in hepatocytes; high expression of GP73 promotes HBV replication and hepatocellular carcinoma stem cell production. It has been reported that NF-kappa B is closely related to the occurrence of hepatocellular carcinoma. GP73 inhibits cellular innate immunity by negatively regulating NF-kappa B. HBV replication also inhibits the innate immunity of hepatocytes. Accordingly, we propose a scientific hypothesis: preS1 encoded by HBV can negatively regulate the expression of NF-kappa B by up-regulating GP73, which results in the suppression of cellular innate immunity and the initiation of HBV-related hepatocellular carcinoma. To confirm the above hypothesis, this project intends to further elucidate the role and mechanism of preS1 up-regulation of GP73 expression in HBV-related hepatocellular carcinogenesis at molecular, cellular and animal levels by using of immunoblotting, fluorescence quantification, flow cytometry, RNA interference and tumorigenesis in nude mice. The successful implementation of this study will elaborate the molecular mechanism of the occurrence and development of HBV-related hepatocellular carcinoma from a new perspective, and provide theoretical support for clinical prevention and treatment of HBV-induced hepatocellular carcinoma.

HBV慢性感染极易发展为肝癌，严重威胁中国1.2亿HBV慢性携带者健康。目前，学术界对HBV相关性肝癌发生的分子机制仍缺乏完整了解。我们前期研究发现：HBV编码的preS1可上调肝细胞GP73表达；GP73高表达可促进HBV复制和肝癌干细胞产生。而有报道，NF-κB与肝癌发生密切相关。GP73通过负调控NF-κB抑制细胞天然免疫。HBV复制也会抑制肝细胞天然免疫。据此，我们提出科学假说：HBV编码的preS1通过上调GP73表达负调控NF-κB，导致细胞天然免疫受抑制，引发HBV相关性肝癌。为证实以上假说，本项目拟进一步在分子、细胞和动物水平，利用免疫印迹、荧光定量、流式细胞术、RNA干扰和裸鼠成瘤等实验手段，阐明preS1上调GP73表达在HBV相关性肝癌发生中所发挥的作用和机制。本研究的顺利实施将从新的视角阐述HBV相关性肝癌发生发展的分子机制，为临床防治HBV引起的肝癌提供理论支撑。

乙型肝炎病毒（Hepatitis B virus, HBV）在临床上会造成急性或是慢性感染。HBV慢性感染与肝癌的发生密切相关。高尔基蛋白73（Golgi protein 73, GP73）在HBV感染和肝癌发生过程中异常升高。但是，GP73在HBV感染和肝癌发生过程中所起的作用机制仍然不清楚。. 本项目研究了GP73在HBV相关性肝癌组织中的表达差异，使用免疫组化、ELISAs, Western blotting 和qRT-PCR等技术分析了HBV感染条件下的肝癌细胞模型和原代肝细胞模型。构建了GP73稳定过表达细胞系，并使用流式细胞术、克隆形成实验、划痕实验、裸鼠成瘤实验等技术分析了细胞模型成瘤能力的差异。最后，我们使用Western blotting和qRT-PCR技术检测了HBV感染和GP73异常上调对细胞天然免疫信号通路的影响。. 我们的实验结果显示：临床HBV阳性肝癌患者的癌组织和血液标本中GP73均有显著升高。使用HBV感染肝细胞模型，可以上调GP73的表达水平。在肝细胞模型中强制过表达GP73可以诱导细胞产生癌症干细胞，增强肝癌细胞的成瘤能力。在HBV感染促进GP73的表达的同时，细胞的天然免疫处于受抑制状态。. 综上所述，我们的研究证明了HBV通过上调GP73实现对细胞天然免疫的抑制，最终导致肝癌的发生。这项研究深入阐释了HBV感染导致肝癌发生的内在机制，并为临床检测和治疗HBV相关性肝癌提供了潜在的作用靶点。