癌细胞侵袭运动、浸润导致的远处转移是结直肠癌（CRC）相关死亡的最大风险因素。我们发现，泛素连接酶TRIM65在CRC中表达上调且与病人复发转移状态相关；干扰TRIM65表达可改变CRC细胞侵袭运动能力，而ARHGAP35是其泛素化底物，但调控细胞运动能力的相关分子机制不详。本项目拟先明确TRIM65与ARHGAP35的蛋白互作，通过片段缺失突变鉴定关键的氨基酸序列；检测下游Rac1/Cdc42/RhoA三个骨架调控主要Rho GTPase的活化状态，观察细胞骨架变化和Rho在细胞中的动态分布；进一步，利用体外细胞和体内转移动物模型，观察TRIM65对CRC细胞骨架和运动转移能力的影响。同时，收集CRC患者标本和资料，探讨TRIM65表达与CRC患者转移及预后的关系。本项目的完成，将阐明新的TRIM65参与细胞骨架和运动转移的分子调控机制，可能为CRC的诊治提供新策略。

Distant metastasis of cancer cells is the most fatal risk factor for colorectal cancer (CRC)-related death. We found that ubiquitin ligase TRIM65 is up-regulated in CRC tissues and is significantly associated with recurrence and metastasis in patients. Pre-experiments indicate that interference with TRIM65 expression may affect ubiquitination of ARHGAP35 and protein accumulation in cells, but the underlining molecular mechanism leading to cell movement and metastasis remains unknown. This project intends to first clarify the physical interaction between TRIM65 and ARHGAP35, to identify key amino acid sequences by fragment deletion mutation; to detect the activation status of three Rho GTPases Rac1/Cdc42/RhoA, and to observe the changes of cytoskeleton and the dynamics of Rho GTPase in cells; furthermore, in vitro cell-based and in vivo transfer animal models are used to observe the effect of different fragmental deletions of TRIM65 on CRC cytoskeleton and motor metastasis ability. Lastly, CRC patient specimens and data are collected to investigate the relationship between TRIM65 expression and metastasis and prognosis in CRC patients. The completion of this project will clarify the novel mechanism and clinical significance of the TRIM65 regulation of cell cytoskeleton and metastasis, and may provide a new strategy for the diagnosis and treatment of CRC.