NEDD9与AGO2在肾细胞癌中相互作用的鉴定及机制研究

Renal cell carcinoma (RCC) is the most lethal of all genitourinary malignancies. The current project aims to study the mechanism of NEDD9 promoting RCC progression through interacting with RNA silencing associated protein-AGO2. Our previous study demonstrated that NEDD9 is significantly up-regulated in RCC compared to the adjacent normal renal tissues. And NEDD9 silencing led to significantly inhibited cell proliferation, increased apoptosis, and cell-cycle arrest in G1 phase. However, the underlying mechanism remains uncovered. Our preliminary experiments using Immunoprecipitation coupled with mass spectrometry identified the potential interaction between NEDD9 and AGO2. Based on above, the current project aims to study the interaction between NEDD9 and AGO2 through Co-IP,FRET assay, illustrating the reciprocal regulation of NEDD9 and AGO2, and their downstream signaling pathway to drive malignant progression of RCC. Our study will contribute to the mechanistic study of the pathogenesis of RCC, and provide novel potential therapeutic targets for RCC targeted therapy.

肾细胞癌（RCC）是泌尿系统最常见的恶性肿瘤之一，然而发病机制不明。本课题旨在研究NEDD9与RNA沉默相关蛋白AGO2相互作用促进RCC恶性侵袭的机制。我们的前期研究发现，NEDD9在RCC中表达显著上调，在肾癌细胞系中敲除NEDD9表达会显著抑制细胞增殖、促进细胞凋亡、并阻滞细胞周期于G1期，而NEDD9的促癌机制尚不明确。预实验中我们通过免疫共沉淀-质谱测定发现NEDD9可能与AGO2相结合。基于上述研究，本课题拟深入探究NEDD9与AGO2之间的相互作用，通过Co-IP,FRET等实验，从体内、体外两个层面系统阐述NEDD9与AGO2之间的相互调控j作用及其促进肾癌恶性进展的下游信号通路的机制，从而为揭示RCC的发病机制提供理论依据，并为RCC的分子靶向治疗 提供新的潜在作用靶点。

肾细胞癌（RCC）约占肾脏恶性肿瘤的90%，对药物治疗不敏感且预后不良，因此，探索新的治疗靶点对肾细胞癌的治疗至关重要。本研究发现NEDD9在RCC组织中表达显著上调，且与患者的不良预后相关。通过细胞划痕、tranwell、MTT及细胞周期、细胞凋亡实验，证实敲除NEDD9抑制RCC细胞的增殖、侵袭和迁移。在动物水平通过皮下成瘤实验，证实敲低NEDD9后小鼠肿瘤的生长受到明显抑制。进一步通过免疫共沉淀、细胞共定位实验，验证了NEDD9与AGO2蛋白之间的相互作用，阐明AGO2的表达受NEDD9调控，并细化构建了NEDD9和AGO2的缺失突变体。本研究为进一步研究NEDD9的相互作用蛋白网络的建立提供依据，对揭示NEDD9基因及AGO2蛋白对肾细胞癌的发生、发展及靶向基因治疗提供依据和新的线索具有重要的临床意义。