微小RNA-425调控重组人Dickkopf相关蛋白3对膀胱癌T24细胞生物学行为的影响  

Effect of microRNA-425 on biological function in bladder cancer T24cells by targeting Dickkopf-related protein 3

作　　者：宁金卓[1] 程帆[1] 余伟民[1] 饶婷[1] 李浩勇[1] 阮远[1] NING Jinzhuo;CHENG Fan;YU Weimin;RAO Ting;LI Haoyong;RUAN Yuan(Dept.of Urology,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei,China)

机构地区：[1]武汉大学人民医院泌尿外科,湖北武汉430060

出　　处：《武汉大学学报（医学版）》2022年第2期233-237,共5页Medical Journal of Wuhan University

摘　　要：目的:探讨微小RNA(miR)-425靶向调控重组人Dickkopf相关蛋白3(DKK3)在膀胱癌细胞中对凋亡、迁移及侵袭的影响。方法:选取2017年1月—2019年8月武汉大学人民医院收集的32例膀胱癌组织及癌旁组织样本作为研究对象,采用qRT-PCR法检测膀胱癌组织及癌旁组织中miR-425表达水平;在膀胱癌T24细胞中采用慢病毒建立对照miRNA和miR-425敲降细胞株(NC inhibitor组和miR-425 inhibitor组),采用Western Blot和qRTPCR法检测细胞中DKK3的蛋白及mRNA表达水平,采用生物信息学和荧光素酶报告实验测定miR-425和DKK3的相关性,分别采用双染法流式细胞术、划痕实验、Transwell实验观察细胞凋亡、迁移和侵袭水平的改变。组间比较采用t检验。结果:miR-425在膀胱癌组织中的表达水平(4.29±0.32)高于癌旁正常组织(1.32±0.21),差异有统计学意义(t=43.895,P<0.05)。miR-425表达水平与肿瘤的组织学分级和肿瘤直径相关(P<0.05)。生物信息学和荧光素酶报告实验验证了DKK3是miR-425的下游靶基因。与NC inhibitor组DKK3蛋白(0.23±0.04)和mRNA(1.00±0.18)表达水平相比,miR-425 inhibitor组DKK3蛋白(0.72±0.06)和mRNA(3.21±0.25)表达水平明显增加,差异有统计学意义(t=11.769,P<0.05;t=12.426,P<0.05)。与NC inhibitor组[凋亡率(10.70±1.34)%]相比,miR-425 inhibitor组凋亡水平[(25.20±1.89)%]显著增加,差异有统计学意义(t=10.840,P<0.05)。与NC inhibitor组相比,miR-425 inhibitor组迁移和侵袭数量显著降低,差异有统计学意义(分别为:t=-4.745,P<0.05;t=-9.784,P<0.05)。结论:miR-425在膀胱癌T24细胞中通过靶向DKK3表达,影响细胞凋亡、增殖和侵袭能力。Objective: To investigate the effect of microRNA(miR)-425 on the apoptosis, migration and invasion in bladder cancer cells by targeting Dickkopf-related protein 3(DKK3). Methods: From January2017 to August 2019, 32 samples of bladder cancer tissue and the adjacent normal tissue were collected from the Renmin Hospital of Wuhan University. qRT-PCR method was used to detect the expression of miR-425 in bladder cancer tissues and adjacent normal tissues. The control miRNA and miR-425 knockdown cell lines(NC inhibitor group and miR-425 inhibitor group) were established in bladder cancer T24 cells by lentivirus. Western Blot and qRT-PCR method were used to examine the protein and mRNA expression level of DKK3. The correlation between miR-425 and DKK3 was measured by bioinformatics and luciferase reporter assay. AnnexinV-FITC/PI, wound healing, and Transwell method were used to detect the changes of apoptosis, migration, and invasion. T test was used for comparison between groups. Results: The expression level of miR-425 in bladder cancer tissues was(4. 29±0. 32) was higher than that of the adjacent normal tissue(1. 32±0. 21), with significant difference(t=43. 895, P<0. 05). The expression of miR-425 was correlated with histological grade and tumor diameter(P<0. 05). Bioinformatics and luciferase reporter assay predicted and verified DKK3 as a direct target of miR-425. Compared with that respectively in NC inhibitor group, DKK3 protein and mRNA expression in miR-425 inhibitor group were significantly increased(0. 23±0. 04 vs0. 72±0. 06, t=11. 769, P<0. 05;1. 00±0. 18 vs 3. 21±0. 25, t=12. 426, P<0. 05). Compared with that in NC inhibitor group, the apoptosis level of miR-425 inhibitor group was significantly increased, as(10. 70±1. 34)% vs(25. 20±1. 89)%(t=10. 840, P<0. 05). There were significant difference between NC inhibitor group and miR-425 inhibitor group in the cell number of migration and invasion(t=-4. 745, P<0. 05;t=-9. 784, P<0. 05). Conclusion: miR-425 could change the apoptosis, migration, and inva

关 键 词：膀胱癌 微小RNA-425 重组人Dickkopf相关蛋白3 凋亡 迁移 侵袭 

分 类 号：R737[医药卫生—肿瘤]