lncRNA MGP-AS2通过miR-18a/Wnt/β-catenin轴抑制膀胱癌的进展  

lncRNA MGP-AS2 Inhibits the Progression of Bladder Cancer Through the miR-18a/Wnt/β-catenin Axis

作　　者：胡国森 杨凌博 李小辉 盖强强 HU Guosen;YANG Lingbo;LI Xiaohui(Department of Urology,Luoyang Central Hospital,Zhengzhou University,Henan 471009,China)

机构地区：[1]郑州大学附属洛阳中心医院泌尿外科,471009 [2]华中科技大学附属同济医院泌尿外科,武汉430033

出　　处：《医学研究杂志》2024年第9期38-44,共7页Journal of Medical Research

基　　金：国家自然科学基金资助项目(82103284);中国博士后科学基金资助项目面上项目(2019M662640)。

摘　　要：目的探讨膀胱癌组织中长链非编码RNA(long non-coding RNA,lncRNA)MGP-AS2表达与膀胱癌患者生存期的关系,研究MGP-AS2对膀胱癌细胞生物学功能的影响及可能机制。方法采用TANRIC数据库分析膀胱癌组织中MGP-AS2表达及其与患者生存期的关系。实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)法检测MGP-AS2在膀胱癌细胞系中的表达水平。将MGP-AS2过表达质粒和阴性对照(negative control,NC)质粒分别转染至膀胱癌5637细胞,分别作为MGP-AS2组和NC组。平板克隆实验、细胞划痕实验以及Transwell实验分别分析过表达MGP-AS2对膀胱癌5637细胞增殖、迁移和侵袭的影响。双荧光素酶报告基因实验验证MGP-AS2和miR-18a之间的靶向结合。TANRIC数据库分析膀胱癌组织中MGP-AS2和miR-18a表达的相关性。RT-qPCR法检测过表达MGP-AS2对膀胱癌5637细胞中miR-18a表达的影响。Western blot法检测过表达MGP-AS2对膀胱癌5637细胞中Wnt/β-catenin信号通路蛋白表达的影响。结果膀胱癌组织中MGP-AS2表达水平显著低于正常膀胱组织(P<0.01)。MGP-AS2高表达患者的总生存期和无病生存期均显著长于MGP-AS2低表达患者(P<0.01)。膀胱癌细胞系中MGP-AS2表达水平均低于SV-HUC-1细胞(P<0.05),MGP-AS2表达最低的膀胱癌细胞是5637细胞(P<0.01)。与NC组比较,过表达MGP-AS2可显著抑制膀胱癌5637细胞的增殖、迁移以及侵袭能力(P<0.01)。双荧光素酶报告基因实验证实MGP-AS2可靶向结合miR-18a(P<0.01)。与NC组比较,过表达MGP-AS2可下调miR-18a的表达(P<0.01)。过表达MGP-AS2后,膀胱癌5637细胞中Wnt/β-catenin信号通路蛋白均下调(P<0.01)。结论膀胱癌中MGP-AS2呈低表达,MGP-AS2表达量与膀胱癌患者的预后相关,MGP-AS2可能通过吸附miR-18a抑制Wnt/β-catenin信号通路转导,从而抑制膀胱癌的进展。Objective To explore the relationship between the expression of lncRNA MGP-AS2 in bladder cancer tissues and the survival of bladder cancer patients,and to study the impact of MGP-AS2 on the biological function of bladder cancer cells and possible mechanism.Methods The TANRIC database was used to analyze MGP-AS2 expression level in bladder cancer tissues and its relationship with patient survival.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of MGP-AS2 in bladder cancer cell lines.The MGP-AS2 overexpression plasmid and negative control(NC)plasmid were transfected into bladder cancer 5637 cells,respectively,as the MGP-AS2group and NC group.Plate cloning experiment,cell scratch experiment and Transwell experiment were used to analyze the effects of MGP-AS2 overexpression on the proliferation,migration and invasion of bladder cancer 5637 cells.Dual-luciferase reporter gene experiment was used to verify the targeted binding between MGP-AS2 and miR-18a.The TANRIC database was used to analyze the correlation between MGP-AS2 and miR-18a expression.RT-qPCR method was used to analyze the effect of MGP-AS2 overexpression on the expression of miR-18a in bladder cancer 5637 cells.Western blot method was used to detect the effect of MGP-AS2 overexpression on the expression of Wnt/β-catenin signaling pathway proteins in bladder cancer 5637 cells.Results The expression level of MGP-AS2 in bladder cancer tissue was significantly lower than that in normal bladder tissues(P<0.01).The overall survival and disease-free survival of patients with high expression of MGP-AS2 were significantly longer than those of patients with low expression of MGP-AS2(P<0.01).The expression level of MGP-AS2 in bladder cancer cell lines were lower than those in SV-HUC-1 cells(P<0.05).The bladder cancer cells with the lowest MGP-AS2 expression was 5637 cells(P<0.01).Compared with the NC group,overexpression of MGP-AS2 could significantly inhibit the proliferation,migration and invasion ability of 5637

关 键 词：膀胱癌 MGP-AS2 miR-18a 细胞增殖 细胞迁移 细胞侵袭 

分 类 号：R544[医药卫生—心血管疾病]