RNA甲基化酶METTL14介导lnc-LSG1 m6A修饰抑制肾透明细胞癌转移的作用及机制研究

肾癌转移机制尚不明确。我们前期发现：METTL14在肾透明细胞癌(ccRCC)低表达，且其水平与恶性程度及预后负相关；细胞及动物实验表明METTL14抑制ccRCC转移；MeRIP测序及功能验证表明METTL14可调控lnc-LSG1 m6A修饰抑制ccRCC转移。lnc-LSG1可与调控FGFR2剪接的ESRP2结合。因此我们提出：METTL14可调控lnc-LSG1的m6A修饰影响其与ESRP2的结合从而改变FGFR2的剪接，最终抑制ccRCC转移。本课题拟综合运用RNA pull-down、RNA-IP、缺失作图分析、裸鼠原位及肺转移肾癌模型等技术，从细胞、动物、临床标本多个层次深入研究METTL14介导的lnc-LSG1 m6A修饰对ccRCC转移的影响并阐明机制。本项目将丰富lncRNA的作用模式，并阐明RNA甲基化修饰调控lncRNA功能的具体方式，为转移性肾癌治疗提供新思路。

At present, there is no effective treatment for metastatic renal cell carcinoma (RCC), and the mechanism of RCC metastasis still needs to be further explored. Our previous study found that: The expression of METTL14 was lower in ccRCC compared to para-carcinoma tissue, and the amount of METTL14 was negatively correlated with the degree of malignancy and prognosis of ccRCC. In vitro and in vivo experiments showed that METTL14 suppressed ccRCC metastasis. MeRIP sequence was performed and the result was verified by PCR, suggesting that METTL14 could regulate the m6A modification of lnc-LSG1 to modulate the metastatic capacity of ccRCC. Lnc-LSG1 could bind to ESRP2, which regulates FGFR2 splicing. Therefore, we hypothesized that METTL14 could regulate the m6A modification of lnc-LSG1, thereby adjusted the splicing of FGFR2 gene via interacting with ESRP2, and ultimately inhibited the ccRCC metastasis. Based on our previous study, this project aims to study the inhibitory effect and underlying mechanism of METTL14-mediated m6A modification of lnc-LSG1 on ccRCC metastasis with cells, animal models and clinical specimens by using RNA-pulldown, RNA-IP, deletion mapping analysis, in situ and lung metastatic ccRCC models etc. This project will enrich the action model of lncRNA, elucidate the specific regulation of RNA function by RNA methylation, and provide new ideas for the treatment of metastatic ccRCC.