rs4969170GG基因型抑制SOCS3基因转录活性促进肝癌发生发展的功能机制研究

本课题组前期研究证实rs4969170A/G SNP的rs4969170GG基因型下调SOCS3的表达，并与肝癌的发生发展、临床病理特征及预后密切相关，但其分子机制尚不清楚。本课题拟利用CRISPR-Cas 技术获得rs4969170A/G不同基因型肝癌细胞，比较其增殖、调亡、细胞周期、克隆形成、迁移、侵袭能力及体内成瘤的差异，明确rs4969170GG促进肝癌发生发展的生物学功能；构建rs4969170GG基因型的knock-in小鼠研究SOCS3 rs4969170GG基因型的成瘤能力。利用双荧光报告系统等技术探讨rs4969170GG对SOCS3基因的转录调控机制；利用动物及HCC样本研究rs4969170GG引起HCC有关信号通路的变化。通过本项目阐明rs4969170GG促进肝癌发生发展的分子机制，为针对SOCS3基因 rs4969170A/G多态性的肝癌个性化治疗提供理论基础。

Our previous studies demonstrated that SOCS3 rs4969170GG genotype was closely related to the occurrence of HCC, the degree of malignancy and the prognosis of HCC. rs4969170GG genotype could reduce the expression of SOCS3 by transcriptional repression of SOCS. However, the molecular mechanism of the occurrence and development of HCC is still unclear. In this study, we obtained rs4969170A/G different alleles of HCC cells using CRISPR-Cas technology. We will reveal the biological function of rs4969170GG in HCC, by comparing the proliferation, apoptosis, cell cycle, clone formation, migration and in vivo tumorigenicity of the different rs4969170A/G alleles HCC cells. We will study the ability of the tumor formation using the rs4969170GG knock-in mice. We will reveal the mechanism of rs4969170GG in regulating the transcriptional activity of the promoter of SOCS3 using luciferase reporter system, Gel shift Assay and Chromatin Immunoprecipitation Assay. Furthermore, we will detect the expression levels of SOCS3 in hepatocellular carcinoma tissue microarray to analyse their relevance and verify the regulation mechanism of SOCS3 on HCC. We will also reveal the association between the related cell signal pathway and SOCS3 rs4969170A/G SNP using our abundant clinical patient resource. This study will reveal the mechanisms of SOCS3 rs4969170GG genotype in promoting the occurrence of HCC and will be helpful to assess the feasibility of SOCS3 as a potential therapeutic target for HCC therapy.