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.
出处
《中华实验外科杂志》
CSCD
北大核心
2017年第12期2054-2056,共3页
Chinese Journal of Experimental Surgery