微小RNA-93靶向大型肿瘤抑制因子2对肾癌细胞GRC-1增殖及凋亡的影响  被引量:6

microRNA-93 regulate proliferation and apoptosis of renal cancer cells by targeting large tumor suppressor homolog 2

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作  者:冯子煜[1] 顾朝辉[1] 李帅[1] 张学冲[1] 兰东阳 刘秉乾[1] 王义昆[1] 贾占奎[1] 杨锦建[1] 武玉东[1] 

机构地区:[1]郑州大学第一附属医院泌尿外科,450052

出  处:《中华实验外科杂志》2017年第12期2054-2056,共3页Chinese Journal of Experimental Surgery

摘  要:目的观察微小RNA(miRNA,miR)-93靶向大型肿瘤抑制因子2(LATS2)对肾癌细胞GRC-1增殖及凋亡的影响。方法选取人肾癌细胞GRC-1,双荧光素酶报告基因实验验证miR-93与LATS2的靶向关系。分别转染Neg-miR、pre-miR-93、anti-miR-93至肾癌细胞GRC-1,采用反转录-聚合酶链反应(RT-PCR)法检测miR-93表达,Western blot检测LATS2蛋白表达,噻唑蓝(MTT)法检测细胞增殖活性,流式细胞仪检测细胞凋亡。结果miR-93与LATS2存在结合位点。与转染Neg-miR比较,共转染pre-miR-93与LATS2-wt可使GRC-1荧光素酶活性降低(P=0.000)。与Neg-miR组比较,pre-miR-93组miR-93表达上调,LATS2蛋白表达降低(P=0.000);anti-miR-93组miR-93表达降低,LATS2蛋白表达增高(P=0.000)。与Neg-miR组比较,pre-miR-93组细胞增殖活性增高,同时凋亡率降低(P=0.000);anti-miR-93组细胞增殖活性降低,同时凋亡率增高(P=0.000)。结论miR-93可通过靶向LATS2调控肾癌细胞增殖及凋亡。ObjectiveTo evaluate the effect of microRNA (miRNA, miR)-93 regulate proliferation and apoptosis of renal cancer cells by targeting large tumor suppressor homolog 2 (LATS2).MethodsChose renal cancer cells line GRC-1, and used dual-luciferase report gene experiment to verification the targeted relationship between miR-93 and LATS2. Respectively transfected Neg-miR, pre-miR-93 and anti-miR-93 to GRC-1 cells. The miR-93 expression was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The LATS2 protein expression was detected by Western blotting. The cell proliferation activity was detected by methyl thiazol tetrazolium (MTT) method. The cell apoptosis were detected by flow cytometry.ResultsBetween miR-93 and LATS2 had some binding sites. Compared with transfection Neg-miR, co-transfection pre-miR-93 and LATS2-wt can made luciferase activity of GRC-1 significantly decreased (P=0.000). Compared with Neg-miR group, the miR-93 expression of pre-miR-93 group significantly increased (P=0.000), and the LATS2 protein expression significantly decreased (P=0.000). The miR-93 expression of anti-miR-93 group significantly decreased (P=0.000), and the LATS2 protein expression significantly increased (P=0.000). Compared with Neg-miR group, the proliferation activity of pre-miR-93 group significantly increased (P=0.000), and the apoptosis rate of significantly decreased(P=0.000). The proliferation activity of anti-miR-93 group significantly decreased(P=0.000), and the apoptosis rate of significantly increased (P=0.000).ConclusionmiR-93 can regulate proliferation and apoptosis of renal cancer cells by targeting LATS2.

关 键 词:微小RNA-93 大型肿瘤抑制因子2 肾癌 增殖 脱噬作用 

分 类 号:R737.11[医药卫生—肿瘤]

 

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