miR-3605-5p通过调控TRPV4表达对肾细胞癌A498细胞侵袭和增殖的影响  

Effect of miR-3605-5p on the invasion and proliferation of renal cell carcinoma A498 cells by regulating TRPV4 expression

作　　者：蔡亮 李超[1] 张林[1] 朱保平[1] CAI Liang;LI Chao;ZHANG Lin;ZHU Baoping(Dept.of Urology,Zhongnan Hospital of Wuhan University,Wuhan 430071,Hubei,China)

机构地区：[1]武汉大学中南医院泌尿外科,湖北武汉430071

出　　处：《武汉大学学报（医学版）》2021年第2期272-276,共5页Medical Journal of Wuhan University

摘　　要：目的:观察微小RNA(miRNA,miR)-3605-5p通过调控瞬时感受器电位离子通道香草素受体4(TRPV4)基因表达对肾细胞癌A498细胞侵袭和增殖的影响。方法:实时定量聚合酶链反应(qRT-PCR)检测肾细胞癌细胞系(A498、CaKi-1、786-O、ACHN)和永生化近端肾小管上皮细胞(HK-2)中miR-3605-5p的表达。将miR-3605-5p表达最少的肾细胞癌细胞系随机分为2组:miR-NC组(转染miR-NC)和miR-3605-5p组(转染miR-3605-5p)。qRT-PCR检测转染效果。Transwell实验和细胞计数试剂盒(CCK-8)检测两组肾细胞癌A498细胞侵袭和增殖水平。生物信息学软件预测和双荧光素酶报告实验验证miR-3605-5p的靶基因。qRT-PCR和Western Blot检测2组肾细胞癌A498细胞中靶基因的表达。结果:miR-3605-5p在肾细胞癌细胞系的表达水平明显低于永生化近端肾小管上皮细胞(P<0.05),在A498细胞中表达最少(P<0.01)。miR-3605-5p组A498细胞中miR-3605-5p的表达(9.49±1.09)高于miR-NC组(1.06±0.18),差异有统计学意义(P<0.01)。miR-NC组和miR-3605-5p组侵袭细胞数量分别为(125.70±14.05)个和(42.01±8.98)个,miR-3605-5p组A498细胞侵袭能力下降(P<0.01)。与miR-NC组比较,miR-3605-5p组A498细胞增殖能力降低(P<0.05)。生物信息学软件显示miR-3605-5p的靶基因是TRPV4。双荧光素酶报告实验证明miR-3605-5p可靶向结合TRPV4基因(P<0.05)。与miR-NC组比较,miR-3605-5p组A498细胞中TRPV4的表达降低(P<0.05)。结论:miR-3605-5p通过下调TRPV4基因表达抑制肾细胞癌细胞A498的侵袭和增殖能力,miR-3605-5p可能成为肾细胞癌的潜在治疗靶点。Objective: To observe the effect of microRNA(miRNA, miR)-3605-5 p on the invasion and proliferation of renal cancer A498 cells by regulating the expression of transient receptor potential vanilloid receptor 4(TRPV4) gene.Methods: Real-time quantitative polymerase chain reaction(qRT-PCR) was used to detect miR-3605-5 p expression in renal cell carcinoma cell lines(A498, CaKi-1,786-O, ACHN) and immortalized proximal tubular epithelial cells(HK-2). Renal cancer cell lines with minimal expression of miR-3605-5 p were randomly divided into two groups: miR-NC group(transfected with miR-NC) and miR-3605-5 p group(transfected with miR-3605-5 p). The transfection effect was detected by qRT-PCR. Transwell assay and cell counting kit(CCK-8) were used to detect the invasion and proliferation of renal cancer A498 cells. Bioinformatics software prediction and dual luciferase reporter assays validated the target gene of miR-3605-5 p. qRT-PCR and Western Blot were used to detect the expression of target genes in the two groups of renal cancer A498 cells.Results: The expression level of miR-3605-5 p in renal cancer cell lines was significantly lower than that in immortalized proximal tubular epithelial cells(P<0. 05), and was the least in A498 cells(P<0. 01). The expression of miR-3605-5 p of A498 cells in miR-3605-5 p group was higher(9. 49±1. 09)than that in miR-NC group(1. 06±0. 18) with significant difference(P<0. 01). The number of invasion cells in miR-NC group and miR-3605-5 p group was(125. 70±14. 05) and(42. 01±8. 98), respectively, indicating that the invasive ability of A498 cells in the miR-3605-5 p group was decreased(P<0. 01). Compared with that in miR-NC group, the proliferation of A498 cells in miR-3605-5 p group was decreased(P<0. 05). The target gene for miR-3605-5 p shown by bioinformatics software is TRPV4. The dual luciferase reporter assay demonstrated that miR-3605-5 p could target the TRPV4 gene(P<0. 05). Compared with that in the miR-NC group, the expression of TRPV4 decreased in the miR-3605-5 p grou

关 键 词：肾细胞癌 微小RNA 瞬时感受器电位离子通道香草素受体4 细胞侵袭 细胞增殖 

分 类 号：R737.11[医药卫生—肿瘤]