膀胱癌组织、细胞中长链非编码RNAs H19表达变化及其对细胞增殖的影响和机制  

Expression changes of LncRNAs H19 in bladder cancer tissues and cells and its effect on proliferation of bladder cancer cells

作　　者：郭宗华[1] 孔东波[1] 江波涛[1] 邹伟[1] 饶志刚[1] 赵克栋[1] GUO Zonghua;KONG Dongbo;JIANG Botao;ZOU Wei;RAO Zhigang;ZHAO Kedong(Xianning Central Hospital,the First Affiliated Hospital of Hubei University of Science and Technology,Xianning 437100,China)

机构地区：[1]咸宁市中心医院湖北科技学院附属第一医院

出　　处：《山东医药》2019年第26期27-30,共4页Shandong Medical Journal

基　　金：湖北省卫生和计划生育委员会科研重点项目(JX6A45);湖北省咸宁市科技局科技计划项目(201747)

摘　　要：目的观察长链非编码RNAs(LncRNAs)H19在膀胱癌组织及细胞中的表达变化,并探讨其对膀胱癌细胞增殖的影响及机制。方法收集40例膀胱癌组织、20例正常膀胱组织,培养人膀胱癌细胞EJ、T24、J82和人正常膀胱上皮细胞SV-HUC-1,用qRT-PCR法检测组织细胞中的H19,并分析H19与膀胱癌患者临床病理特征的关系。将H19表达最高的膀胱癌细胞随机分为抑表达组和对照组,用脂质体2000分别转染si-H19、si-NC;转染48 h后收集细胞,采用MTS法测算细胞增殖率、平板克隆实验观察细胞克隆形成能力、流式细胞仪检测细胞周期、Western blotting法检测细胞中的wnt/β-catenin信号通路相关蛋白β-catenin、Cyclin D1和cMYC。结果膀胱癌组织中H19表达高于正常膀胱组织,T 3~T 4期膀胱癌组织中H19表达高于T 1~T 2期(P均<0.01);膀胱癌细胞EJ、T24、J82中H19表达均高于人正常膀胱上皮细胞SV-HUC-1(P均<0.05),其中T24细胞中H19表达最高;与对照组比较,抑表达组T24细胞中细胞增殖率降低,细胞克隆形成数减少,G 1期细胞比例增加,细胞中β-catenin、Cyclin D1和cMYC蛋白表达减少(P均<0.05)。结论H19在膀胱癌组织和细胞中高表达,且肿瘤分期越高表达越高;H19降表达可以抑制膀胱癌细胞增殖,其机制可能与抑制wnt/β-catenin信号通路相关下游靶基因的表达有关。Objective To observe the expression changes of long-chain non-coding RNAs(LncRNAs)H19 in bladder cancer tissues and cells,and to explore its effect on the proliferation of bladder cancer cells and its mechanism.Methods Forty cases of bladder cancer tissues and 20 cases of normal bladder tissues were collected.Human bladder cancer cells EJ,T24,J82 and human normal bladder epithelial cells SV-HUC-1 were cultured.The expression of H19 in the tissues and cells was detected by qRT-PCR,and the relationship between H19 and the clinical pathological features of bladder cancer patients was analyzed.The expression of H19 in the bladder cancer cell lines EJ,T24,J82 and human normal bladder epithelial cell line SV-HUC-1 were detected by qRT-PCR;the bladder cancer cells with the highest H19 expression were randomly divided into two groups:the inhibition group and control group,which were transfected with si-H19 and si-NC by liposome 2000,respectively;the cells were collected after 48-hour transfection,the cell proliferation rate was measured by MTS method,and the cell clone formation ability was observed by plate cloning experiment.The cell cycle was detected by cytometry,and the wnt/β-catenin signaling pathway-related proteinsβ-catenin,Cyclin D1 and cMYC were detected by Western blotting.Results The expression of H19 in the bladder cancer tissues was higher than that in the normal bladder tissues.The expression of H19 in T 3-T 4 bladder cancer tissues was higher than that in T 1-T 2 phase(P<0.01).The expression of H19 in the bladder cancer cells EJ,T24 and J82 was higher than that in the normal bladder epithelial cells SV-HUC-1(P<0.05),with the highest expression of H19 in T24 cells.Compared with the control group,the cell proliferation rate and the number of cell clone formation in T24 cells decreased,the proportion of cells in the G 1 phase increased,and theβ-catenin,cyclinD1 and cMYC protein expression decreased in the inhibition group(all P<0.05).Conclusion H19 is highly expressed in bladder cancer tissues and cells,a

关 键 词：膀胱癌 长链非编码RNAs H19 细胞增殖 细胞周期 WNT/Β-CATENIN信号通路 

分 类 号：R737.14[医药卫生—肿瘤]