setdb1与Tiam1相互作用通过调控EMT促进肝癌侵袭转移

项目名称： setdb1与Tiam1相互作用通过调控EMT促进肝癌侵袭转移

项目编号： No.81502532

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 黄静

作者单位： 广东医科大学

项目金额： 18万元

中文摘要： Tiam1是我们前期筛选并验证的肝癌转移促进蛋白。我们应用酵母双杂交技术获得一个全新的Tiam1相互作用蛋白setdb1。初步研究结果显示setdb1可以促进肝癌细胞体外增殖和侵袭能力，其在肝癌细胞株的表达趋势与Tiam1一致，并且setdb1可以诱导肝癌细胞EMT、影响肿瘤干细胞特征。基于前期研究结果，我们提出setdb1与Tiam1相互作用通过调控EMT促进肝癌侵袭转移的假说。为验证这一假说，本项目首先探讨setdb1在肝癌中的作用，过表达或沉默setdb1，利用肿瘤体内、外实验探讨其生物学功能。通过GST pull down、免疫共沉淀、荧光共定位等方法，验证两者的相互作用，研究Tiam1与setdb1之间的结合区域，分析相互作用的位点。然后解析setdb1/Tiam1蛋白质相互作用对EMT发生及肝癌侵袭转移的影响和分子机制，以期为临床拮抗肝癌转移提供新的靶点和治疗策略。

中文关键词： C09_肝和肝内胆管肿瘤；setdb1；Tiam1；上皮间质转化；蛋白质相互作用

英文摘要： Tiam1 is a metastasis-promoting protein for hepatocellular carinoma we have screened and verified. Using the Tiam1-C1199 as a bait, We identified a novel Tiam1-interacting protein setdb1. Preliminary study showed that setdb1 expression in hepatocellular carcinoma cell lines is consistent with Tiam1. Overexpression of setdb1 promoted HCC cells proliferation and invasion ability in vitro while knockdown of setdb1 resulted in the reverse. Moreover, setdb1 induced EMT and affected cancer stemness characteristics. Based on those findings, we hypothesized that setdb1 cooperates with Tiam1 to promote hepatocellular carcinoma metastasis by regulating epithelial-mesenchymal transition. To test this hypothesis, the role of setdb1 in HCC will be firstly explored, while setdb1 overexpression and knockdown HCC cell lines are constructed to investigate its biological function. The interaction between the two should be verified by GST pull down, co-immunoprecipitation and fluorescence colocalization analyses. Their binding domain and regulatory mechanism will be further investigated. Finally, the molecular mechanisms that setdb1 cooperates with Tiam1 to promote hepatocellular carcinoma metastasis by regulating epithelial-mesenchymal transition will be discovered. All of the work are aim to provide a new target for HCC metastasis and treatment strategies.

英文关键词： hepatocellular carcinoma;setdb1;Tiam1;EMT;protein interaction