LncRNA SNHG9内源性竞争YWHAG蛋白调控GSK3β/KLC3/β-catenin信号通路抑制膀胱癌转移的机制研究

转移是膀胱癌致死的主要原因，目前对于膀胱癌转移仍缺乏有效的预测和治疗靶点。非编码RNA对靶蛋白的翻译后调控是近年来研究热点，但在膀胱癌转移中的功能知之甚少。我们前期对膀胱癌及转移癌临床样本进行多组学测序，筛选出在膀胱癌中显著下调的lncRNA SNHG9以及KLC3突变基因。进一步功能研究发现，SNHG9通过调控KLC3抑制膀胱癌转移，其可能机制为内源性竞争YWHAG蛋白调控KLC3。据此我们提出假说：SNHG9内源性竞争YWHAG蛋白调控GSK3β/KLC3/β-catenin信号通路抑制膀胱癌转移。本项目将以临床膀胱癌样本、膀胱癌转移细胞系及裸鼠转移瘤模型为研究对象，采用免疫共沉淀、酵母三杂交和小动物活体成像等技术，探究SNHG9和YWHAG对GSK3β/KLC3/β-catenin通路影响，阐明膀胱癌转移的关键驱动基因和分子机制，为膀胱癌转移预警和寻找诊断和治疗靶点提供重要理论依据。

Metastasis is the leading cause of death in patients with bladder cancer. To date, there is no effective target for the prediction and treatment of bladder cancer metastasis. Study on the posttranslational modificationof protein regulated by non-coding RNA has been hot in research. However, the role of non-coding RNA on posttranslational modification in bladder cancer metastasis in unknown. Through high throughput sequencing, the down-regulated lncRNA SNHG9 and KLC3 missense mutation were identified in bladder cancer and metastatic bladder cancer. Further experiments indicated that lncRNA SNHG9 suppressed the bladder cancer metastasis through regulating KLC3, and SNHG9 may inhibit the GSK3β/KLC3/β-catenin pathway via acting as a competitive endogenous RNA of YWHAG protein. Thus, we proposed a hypothesis that lncRNA SNHG9 suppresses bladder cancer metastasis through regulating GSK3β/KLC3/β-catenin pathway by acting as a competitive endogenous RNA of YWHAG protein. This project will be performed in bladder cancer, metastatic bladder cancer cell and nude mice by utilizing numerous experimental techniques including Co-IP, three-binding hybrid system and live imaging of tumor in small animals. The aim of this project is to explore effects of SNHG9 and YWHAG protein on GSK3β/KLC3/β-catenin pathway and reveal the mechanism and the critical driver in bladder cancer metastasis. Results of this study will provide theoretical base for the prediction and the screen of therapeutic targets in bladder cancer metastasis.