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

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

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.