长链非编码RNA RP11-690G19.3作为ceRNA在肾癌粘附、侵袭和转移中的功能及机制研究

Current researches show close relationship between long non-coding RNAs and carcinoma. The probable mechanism is its regulation in miRNA function as a competing endogenous RNA. In the previous work we found increased expressions of long non-coding RNA RP11-690G19.3 in renal carcinomas, and they were even higher in metastases. In contrast,the expressions of miR-381 were lower in renal carcinomas. Stable binding sites between RP11-690G19.3 and miR-381 are showed according to the bioinformatics analysis. It is well known that miR-381 is related to carcinoma metastasis. Interference of RP11-690G19.3 not only resulted in over-expression of miR-381,but also suppressed the proliferation and invasion of cancer cells. Based on the above, we put forward the hypothesis that RP11-690G19.3 combines miR-381 and therefore regulates the metastasis of renal carcinoma. We plan to investigate the roles of RP11-690G19.3 in the adhesion, invasion and metastasis of renal carcinoma. We will figure out the relationship between RP11-690G19.3 and metastasis of renal carcinoma. Then, we will analysis the binding sites of RP11-690G19.3/miR-381 with the mutations before they are validated by means of dual-luciferase reporter assay. Finally, we will explore the downstream target genes and signal pathways by gene expression profiles. It will clearly elucidate the function and mechanism of RP11-690G19.3/miR-381 in the adhesion, invasion and metastasis of renal carcinoma, which will provide a novel target in the treatment of renal carcinoma.

研究表明长链非编码RNA和肿瘤密切相关，其机制可能与它作为竞争性内源性RNA调控miRNA功能有关。前期工作发现长链非编码RNA RP11-690G19.3在肾癌中表达增高，转移灶中更高，miR-381与之相反。生物信息学显示RP11-690G19.3与miR-381有稳定的结合位点，而miR-381已知与肿瘤转移相关。干扰RP11-690G19.3后miR-381表达增加，细胞增殖和侵袭能力下降。由此，我们提出RP11-690G19.3结合miR-381进而调控肾癌转移的新假说。我们拟研究RP11-690G19.3对肾癌粘附、侵袭和转移能力的影响，分析其与肾癌转移的关系；通过突变其结合位点分析其与miR-381作用靶点，并以双荧光素酶实验验证；通过基因表达谱探索RP11-690G19.3/miR-381下游基因和通路，明确其在肾癌粘附、侵袭和转移中的功能及机制，为肾癌治疗提供新靶点。

本项目通过体外实验证实了RP11-690G19.3对肾癌粘附、侵袭和转移能力的影响，通过检测RP11-690G19.3表达研究了其与肾癌转移的关系。通过突变RP11-690G19.3与miR-381的结合位点，分析二者的靶向调控关系，再结合miRNA靶基因生物信息学软件、qRT-PCR和Western Blot进一步筛选和验证RP11-690G19.3/miR-381下游的基因，阐明了RP11-690G19.3调控肾癌粘附、侵袭和转移的机制，最终丰富的肾癌发生、发展的理论。