miR-122通过介导sprouty2调控肾癌细胞的增殖
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国家自然科学基金面上项目(81372757;81171963);江苏省科技厅基础研究计划(自然科学基金) (BK2008473)


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

    目的:研究miR-122在肾癌发生中的作用以及与sprouty2之间的关系。方法:通过RT-qPCR实验,测定32对肾癌组织及邻近癌旁组织中miR-122的表达水平。将miR-122抑制剂(miR-122 inhibitor)转染至肾癌细胞株786-0和CAKI-1中,下调miR-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-qPCR和Western blot检测sprouty2 mRNA和蛋白表达情况。通过双荧光报告素酶基因实验,研究miR-122与sprouty2 3′UTR结合情况。结果:miR-122在肾癌组织中表达明显升高(P < 0.05);miR-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miR-122 inhibitor 能与sprouty2 3′UTR结合,显著升高荧光值(P < 0.05)。结论:miR-122促进肾癌细胞的增殖,sprouty2可能是miR-122的靶基因。

    Abstract:

    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 specimens,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. [Key words] miR-122;sprouty2;renal cell carcinoma;proliferation

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张 井,张 磊,李 双,柳金良,曹 强,秦 超,殷长军. miR-122通过介导sprouty2调控肾癌细胞的增殖[J].南京医科大学学报(自然科学版),2015,(4):517-522

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  • 收稿日期:2014-11-09
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  • 在线发布日期: 2015-05-05
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