MicroRNA-140-5p通过蛋白激酶C亚型ε促进肾癌细胞增殖（英文）  

MicroRNA-140-5p promotes cell proliferation via protein kinase C isoform ε in renal cell carcinoma

作　　者：刘伟红 余肖 任雨 刘开泰 林晨 王萍[1,3] LIU Wei-hong;YU Xiao;REN Yu;LIU Kai-tai;LIN Chen;WANG Ping(Department of Human Anatomy,Histology and Embryology,Medical School,Ningbo University,Zhejiang Ningbo 315211,China;Laboratory Kidney Carcinoma,Ningbo Urology and Nephrology Hospital,Zhejiang Ningbo 315000,China;Ningbo City Medical Treatment Center Lihuili Hospital,Medical School,Ningbo University,Zhejiang Ningbo 315041,China;Laboratory of Kidney Carcinoma,Urology and Nephrology Institute of Ningbo University,Zhejiang Ningbo 315000,China)

机构地区：[1]宁波大学医学院人体解剖学与组织学胚胎学科系,浙江宁波315211 [2]宁波市泌尿肾病医院肾癌实验室,浙江宁波315000 [3]宁波大学泌尿肾病研究所肾癌实验室,浙江宁波315000 [4]宁波大学医学院李惠利医院宁波市医疗中心,浙江宁波315041

出　　处：《解剖学报》2019年第2期185-191,共7页Acta Anatomica Sinica

基　　金：National Natural Science Foundation of China(81372209);Scientific Plan of Medical and Health of Zhejiang Province(2015RCB024);Ningbo Natural Science Foundation(2017A610185,2017A610193);K.C.Wong Magna Fund in Ningbo University

摘　　要：目的探讨microRNA-140-5p(miR-140-5p)在肾癌发生中的作用。方法 Real-time PCR方法检测肾癌细胞(Caki-1、ACHN、OS-RC-2和786-O)中miR-140-5p的表达。MTS,流式细胞术和Western blotting检测细胞增殖,细胞周期和相关蛋白的改变。结果降低Caki-1细胞中miR-140-5p的水平后,细胞增殖被抑制,G0/G1期细胞数量显着增加,cyclin D、cyclin E和CDK2、CDK4也随之减少;此外,增加786-O细胞中miR-140-5p的水平后,促进体外细胞增殖和体内肿瘤发生。基因芯片结果发现,蛋白激酶C亚型ε(PKCε)基因在改变Caki-1和786-O的细胞中显著性升高。si PKCε在降低肾癌细胞(RCC)中PKCε蛋白表达的同时,还能显著性反转抑制物/模拟物诱导的细胞增殖和细胞周期的改变。结论 miR-140-5p通过PKCε促进肾癌细胞增殖。Objective To explore the role of micro RNA-140-5 p( miR-140-5 p) in renal cell carcinoma( RCC)tumorigenesis. Methods The expression of miR-140-5 p was determined by Real-time PCR in RCC( Caki-1,ACHN,OSRC-2 and 786-O),and compared with kidney cortex proximal tubule cell( HK-2). After Caki-1 and 786-O cells were transfected with miR-140-5 p inhibitor or mimic,the cell proliferation,cell cycle and related proteins were evaluated by MTS assay,flow cytometry and Western blotting. Results Caki-1 and 786-O showed the highest and lowest expression of miR-140-5 p respectively. After the level of miR-140-5 p in Caki-1 cells was decreased,cell proliferation was inhibited and cell number in G0/G1 phase was increased significantly,along with the reduction of cyclin D, E and CDK2,4.Furthermore,the increasing level of miR-140-5 p in 786-O cells promoted cell proliferation in vitro and tumorigenesis in vivo. Moreover,microarray demonstrated that protein kinase C isoform ε( PKCε),a markedly enhanced gene in RCC cells with the changed level of miR-140-5 p. The combination treatment with down-regulation of PKCε,inhibitor/mimic induced changes of cell proliferation and cell cycle arrests were marked reversed in Caki-1 and 786-O cells. Conclusion These result indicate that miR-140-5 p may promote cell proliferation via PKCε in RCC.

关 键 词：MicroRNA-140-5p 肾癌细胞 细胞增殖 细胞周期 实时定量聚合酶链反应 人 

分 类 号：R733-36.2[医药卫生—肿瘤]