miR-651和miR-708作为肾细胞癌分子标志物的价值及其作用机制研究

肾细胞癌（RCC）发病率逐年增高，呈年轻化趋势，但无理想的分子诊断标志物。我们发现，miR-651和miR-708在RCC血清、组织及肾癌细胞株均低表达；生物信息学预测它们参与RCC的调控网络，荧光素酶实验证实了RAP1B等原癌基因是直接靶标。据此提出假设：miR-651和miR-708是RCC潜在的分子标志物，它们作为抑癌miRNA参与RCC的发生发展。为验证假说，我们拟通过临床标本、肾癌细胞株及荷瘤小鼠模型，采用低密度芯片、qRT-PCR、Western blot、免疫共沉淀、脂质体及慢病毒载体转染、RNA干扰等手段，确认miR-651和miR-708对RCC的诊断和预后价值；探明它们在RCC发生发展中的作用及分子机制；构建它们与上游转录因子和下游靶基因间的调控网络。力图为实现RCC无创伤性检出和治疗效果适时监控开辟一条新途径，同时也为进一步阐明RCC的发病机制，为临床诊疗提供新思路。

The incidence of renal cell carcinoma (RCC) is increasing annually and is getting younger, however, no specific diagnostic marker has been discovered. Our previous studies found that the levels of miR-651 and miR-708 were significantly downregulated in RCC tissues, cell lines and sera samples from patients with RCC; Results from bioinformatics analysis suggested that miR-651 and miR-708 might be involved in the regulatory network of RCC, and RAP1B and several other oncogenes have been confirmed to be the direct target genes of miR-651 and miR-708 by using luciferase reporter system. Thus, we hypothesize that miR-651 and miR-708 are potential molecular biomarkers of RCC and they may act as tumor suppressors in RCC. In order to verify this hypothesis, we will adopted clinical samples, RCC cell lines and tumor-bearing mouse model and use some techniques such as TaqMan Low Density Chip assay, qRT-PCR, western blot, co-immunoprecipitation, liposomes and lentivirus transfection, RNA interference, etc. to define the diagnostic and prognostic values of miR-651 and miR-708 for RCC; to explore the roles and the mechanisms of miR-651 and miR-708 in the development and progression of RCC; and to construct the regulatory networks between miR-651, miR-708 and their upstream transcription factors and their down target gens. The purpose of our study is to open up a new way to improve the diagnosis and monitoring of RCC and to provide a new approach and theoretical foundation for the pathogenesis of RCC.