TRIM47参与抑制肝癌侵染转移的机理研究

The proteins of TRIM family are becoming hot-spot, which involved in pathological processes of cancer as the RING-finger E3 ligases. Actually, it is still unclear for many members of TRIMs. In this project, the TRIM47 (an unknown members of TRIMs) was identified to be differentially expressed between HCCLM6 and MHCC97L or MHCC97H cell lines though quantification proteomics data analysis. We speculate that the TRIM47 play important roles in the invasion and migration in HCC. For further exploring the mechanisms of the TRIM47, the ubiquitination activity will be detected in vitro. And the differentially expressed TRIM47 will be identified to associate with metastasis of HCC cell lines through cell assay. The method of the Co-IP combined with LC-MS/MS will be used to identify the ubiquination substrates of the TRIM47. The SILAC-based proteomics technology will cover almost the proteins interacted with the TRIM47, which is useful to understand the mechanism of ubiquitin ligase in the metastasis of HCC. This project will give more indication for the TRIM47 as potential drug target in HCC.

TRIM蛋白家族作为泛素连接酶参与癌症发生发展及转移过程已受到高度关注，但该家族中还有许多蛋白的功能未被研究。本课题建立在对不同转移潜能肝癌细胞系蛋白质组学数据分析的基础上，发现TRIM47在HCCLM6细胞中的表达水平显著低于在MHCC97L和MHCC97H细胞中的表达水平，由此推测TRIM47参与了抑制肝癌侵染迁移的过程，但到目前为止对TRIM47的功能研究未见任何报道。因此将首先研究TRIM47在体外是否具有泛素连接酶的活性；其次通过在侵染迁移能力较强的MHCC97H细胞中高表达或敲除TRIM47蛋白，确定TRIM47蛋白参与抑制肝癌侵染迁移的功能；然后采用CO-IP靶向蛋白质组学技术寻找TRIM47直接作用的底物蛋白，同时结合SILAC标记的定量蛋白质组学技术构建TRIM47调控的肝癌侵染迁移信号途径网络；最终通过体内荷瘤实验验证TRIM47在生物体内的功能。

TRIM蛋白家族作为泛素连接酶参与癌症发生发展及转移过程已受到高度关注，但该家族中.还有许多蛋白的功能未被研究。本课题建立在对不同转移潜能肝癌细胞系蛋白质组学数据.分析的基础上，发现TRIM47在HCCLM6细胞中的表达水平显著低于在MHCC97L和MHCC97H细胞.中的表达水平，由此推测TRIM47参与了抑制肝癌侵染迁移的过程，但到目前为止对TRIM47.的功能研究未见任何报道。因此将首先研究TRIM47在体外是否具有泛素连接酶的活性；其.次通过在侵染迁移能力较强的MHCC97H细胞中高表达或敲除TRIM47蛋白，确定TRIM47蛋白.参与抑制肝癌侵染迁移的功能；然后采用CO-IP靶向蛋白质组学技术寻找TRIM47直接作用.的底物蛋白，同时结合SILAC标记的定量蛋白质组学技术构建TRIM47调控的肝癌侵染迁移.信号途径网络；最终通过体内荷瘤实验验证TRIM47在生物体内的功能。