miR-122通过介导sprouty2调控肾癌细胞的增殖

miR-122 regulates cell proliferation by targeting sprouty2 in renal cancer cell lines

目的 :研究mi R-122在肾癌发生中的作用以及与sprouty2之间的关系。方法 :通过RT-q PCR实验,测定32对肾癌组织及邻近癌旁组织中mi R-122的表达水平。将mi R-122抑制剂(mi R-122 inhibitor)转染至肾癌细胞株786-0和CAKI-1中,下调mi R-122的表达,在不同时间点,CCK-8实验检测细胞增殖,流式细胞仪检测细胞周期,RT-q PCR和Western blot检测sprouty2m RNA和蛋白表达情况。通过双荧光报告素酶基因实验,研究mi R-122与sprouty2 3′UTR结合情况。结果 :mi R-122在肾癌组织中表达明显升高(P<0.05);mi R-122 inhibitor转染后能够抑制肾癌细胞增殖(P<0.05)及导致细胞周期阻滞(P<0.05),并促进肿瘤细胞中sprouty2蛋白的表达(P<0.05);双荧光报告素酶基因实验表明mi R-122 inhibitor能与sprouty2 3′UTR结合,显著升高荧光值(P<0.05)。结论:mi R-122促进肾癌细胞的增殖,sprouty2可能是mi R-122的靶基因。

Objective：To determine the biological function of miR-122 in kidney tumors and the relationship between sprouty2 and miR-122. Methods：RT-qPCR was performed to measure the levels of miR-122 in 32 primary renal cell carcinoma （RCC） and their adjacent non-malignant tissue samples. The miR-122 inhibitor was successfully transfected into 786-0 and CAKI-1 cell lines. CCK-8 assays and cell cycle assays were performed to analyze the proliferation after transfection, respectively. The regulative function of miR- 122 on sprouty2 was evaluated by Western blot assays. Dual luciferase reporting assays were performed to confirm whether sprouty2 is a direct target of miR-122. Results：In RCC spccimens,miR-122 was significantly up-regulated （P 〈 0.05）. The inhibition of miR-122 suppressed the proliferation （P 〈 0.05）,induced cell cycle arrest （P 〈 0.05）, and promoted the expression of sprouty2 protein levels （P 〈 0.05） in 786-0 and CAKI-1 cell lines. Sprouty2 was identified as a new target of miR-122 by dual luciferase,and fluorescence value was significantly increased （P 〈 0.05）. Conclusion：miRNA promotes the proliferation of renal cancer cells and sprouty2 gene may be a target of miR-122.