敲减lncRNA RP11-626G11.3通过调控miR-532-3p抑制人肾癌细胞系的增殖和迁移  

Knockdown of lncRNA RP11-626G11.3 inhibits proliferation and migration of human renal carcinoma cell lines through regulating miR-532-3p

作　　者：王文龙 唐一君 王青兵[1] 陈科 程继强 WANG Wenlong;TANG Yijun;WANG Qingbing;CHEN Ke;CHENG Jiqiang(Department of the Fifth Ward of Surgery,Anyang Cancer Hospital Affiliated to Henan University of Science and Technology,Anyang 455000;Department of the Fifth Ward of Medical Oncology,Anyang Cancer Hospital Affiliated to Henan University of Science and Technology,Anyang 455000;Department of Nephrology,Union Hospital Affiliated to Huazhong University of Science and Technology,Wuhan 430022,China)

机构地区：[1]河南科技大学附属安阳市肿瘤医院外科五病区,河南安阳455000 [2]河南科技大学附属安阳市肿瘤医院肿瘤内科五病区,河南安阳455000 [3]华中科技大学附属协和医院肾内科,湖北武汉430022

出　　处：《基础医学与临床》2023年第10期1498-1504,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81772721)。

摘　　要：目的研究长链非编码RNA(lncRNA)RP11-626G11.3在肾癌组织和细胞系中的表达,探究敲减RP11-626G11.3对肾癌细胞生物学行为的影响。方法用GEPIA数据库分析RP11-626G11.3在肾癌组织和癌旁组织中的表达情况,用TCGA数据库分析RP11-626G11.3表达水平与肾癌患者生存期的关系。QPCR检测RP11-626G11.3在多种肾癌细胞系中的表达。选择RP11-626G11.3表达最高的肾癌细胞系,分别转染对照质粒(si-NC组)和RP11-626G11.3沉默质粒(si-RP11-626G11.3组)。MTT法和细胞划痕实验检测细胞的增殖和迁移。通过生物信息学方法找到与RP11-626G11.3结合的微小RNA(miRNA),并用双荧光素酶报告实验进行验证。QPCR检测两组细胞中miR-532-3p的表达。Western blot检测两组细胞中Wnt/β-catenin信号通路蛋白的表达。结果与癌旁组织相比,肾癌组织中RP11-626G11.3表达量上调(P<0.01)。与RP11-626G11.3高表达的患者相比,RP11-626G11.3低表达的患者生存期较长(P<0.01)。与永生化肾小管上皮细胞相比,肾癌细胞系中RP11-626G11.3表达量均上调(P<0.01),OS-RC-2细胞中RP11-626G11.3的表达量最高(P<0.01)。与si-NC组相比,si-RP11-626G11.3组OS-RC-2细胞活力明显降低(P<0.05),细胞迁移率明显降低(P<0.01)。RP11-626G11.3靶向结合miR-532-3p(P<0.01)。相比si-NC组,si-RP11-626G11.3组OS-RC-2细胞中miR-532-3p表达明显上调(P<0.01),Wnt/β-catenin信号通路蛋白β-catenin、Axin2、C-myc、cyclin D1、MMP7表达水平下降。结论RP11-626G11.3在肾癌组织和细胞系中表达量升高,敲减RP11-626G11.3通过调控miR-532-3p表达抑制肾癌细胞系的增殖和迁移。Objective To study the expression of long-chain non-coding RNA(lncRNA)RP11-626G11.3 in renal cancer tissues and cell lines,and to explore the effect of knockdown of RP11-626G11.3 on the biological behavior of renal cancer cells.Methods The GEPIA database was used to analyze the expression of RP11-626G11.3 in renal cancer tissues and adjacent tissues,and the TCGA database was used to analyze the relationship between the expression of RP11-626G11.3 and the survival time of renal cancer patients.The expression of RP11-626G11.3 in various renal cancer cell lines was detected by qPCR.The renal cancer cells with the highest expression of RP11-626G11.3 were selected and transfected with control plasmid(si-NC group)and RP11-626G11.3 silencing plasmid(si-RP11-626G11.3 group).Cell proliferation and migration were examined by MTT assay and cell scratch assay.The microRNA(miRNA)binding to RP11-626G11.3 was found by bioinformatics method,and verified by dual-luciferase reporter gene experiment.The expression of miR-532-3p in the two groups of cells was detected by qPCR.Western blot was used to detect the expression levels of Wnt/β-catenin signaling pathway in the two groups of cells.Results Compared with adjacent tissues,the expression of RP11-626G11.3 was up-regulated in renal cancer tissues(P<0.01).Compared with patients with high expression of RP11-626G11.3,patients with low expression of RP11-626G11.3 had a longer survival time(P<0.01).Compared with immortalized renal tubular epithelial cells,the expression of RP11-626G11.3 was up-regulated in renal cancer cell lines(P<0.01),and the expression of RP11-626G11.3 was the highest in OS-RC-2 cells(P<0.01).Compared with si-NC group,the viability of OS-RC-2 cells in si-RP11-626G11.3 group was significantly decreased(P<0.05)with decreased cell migration rate(P<0.01).RP11-626G11.3 was found to target at miR-532-3p(P<0.01).Compared with the si-NC group,the expression of miR-532-3p in OS-RC-2 cells in the si-RP11-626G11.3 group was significantly up-regulated(P<0.01),and the Wnt/β-

关 键 词：肾肿瘤 RP11-626G11.3 miR-532-3p 增殖 迁移 

分 类 号：R737.11[医药卫生—肿瘤]