LINC00945调控VEGF的m6A甲基化修饰促进肾癌侵袭转移的机制研究

Distant metastasis is a difficult problem in the treatment of renal cell carcinoma (RCC). lncRNA is closely related to RCC metastasis, but the specific mechanism remains unclear. Our previous study found: ①LINC00945 was highly expressed in RCC and positively correlated with the clinical stage and poor prognosis of patients. Vitro experiments confirmed that LINC00945 could promote invasion and metastasis of RCC; ②High-throughput sequencing and related experiments suggested that LINC00945 colud recruit m6A methyltransferase WTAP to up-regulate the expression of VEGF; ③ Bioinformatics analysis showed that the transcription factor FOXD1 could bind to the site of LINC00945 promoter region and activate its transcription. Consequently, we hypothesized that FOXD1 promote transcription of LINC00945, and LINC00945 could enhance invasion and metastasis of RCC by recruiting WTAP to increase the m6A methylation of VEGF and up-regulate its expression. We intend to elucidate the specific mechanism of LINC00945 regulating m6A methylation modification of VEGF to promote invasion and metastasis of RCC in both in vitro and in vivo，which could provide new ideas for the diagnosis and treatment of RCC.

远处转移是肾癌治疗的一大难题。lncRNA与肾癌转移密切相关，但具体作用机制仍不明确。我们前期发现：①LINC00945在肾癌中高表达并与患者临床分期和不良预后正相关，体外实验证实LINC00945可促进肾癌侵袭转移；②高通量测序及相关实验提示LINC00945可能通过招募m6A甲基转移酶WTAP上调VEGF的表达；③生物信息学分析显示LINC00945启动子区存在转录因子FOXD1结合位点，其高表达可能受FOXD1转录调控。由此提出假说：FOXD1促进LINC00945转录，进而LINC00945通过招募WTAP增加VEGF的m6A甲基化修饰水平并上调其表达，促进肾癌侵袭转移。为验证这一假说，本课题拟从临床样本、细胞水平和动物水平三个层面，阐明LINC00945调控VEGF的m6A甲基化修饰促进肾癌侵袭转移的具体机制，以期为肾癌的临床治疗提供新的理论依据。

远处转移是肾癌治疗的一大难题。lncRNA与肾癌转移密切相关，但具体作用机制仍不明确。本课题通过临床样本、细胞水平和动物水平三个层面的研究，发现LINC00945在肾癌中高表达并与患者临床分期和不良预后正相关，并且LINC00945能够在体内外水平促进肾癌的侵袭和迁移能力。机制方面，我们明确了FOXD1促进LINC00945的转录，进而LINC00945通过结合HuR增加VEGF mRNA稳定性并上调其表达，促进肾癌的侵袭转移。最后通过拓展研究，我们发现USP36以ERK2依赖的方式与WTAP相互作用，调节WTAP的去泛素化和稳定性促进肾癌的侵袭转移。本项目的研究结果可为肾癌的临床治疗提供新的理论依据。