hsa-miR-6832-5P抑制膀胱肿瘤细胞生长和转移的作用及机制  

Inhibitory effect and mechanism of hsa-miR-6832-5p on growth and metastasis of bladder tumor cells

作　　者：张茂娜 张弘[1] 王艳莎 吕思佳 Zhang MaonoL;Zhang Hong;Wang Yansha;Lyu Sijia(Department of Pathology,Ezhou Central Hospital,Ezhou 436000,China;Department of Pathology,Jincheng People's Hospital,Jincheng 048026,China)

机构地区：[1]湖北省鄂州市中心医院病理科,436000 [2]山西省晋城市人民医院病理科,048026

出　　处：《中国医师杂志》2020年第9期1376-1380,共5页Journal of Chinese Physician

摘　　要：目的分析人微小RNA-6832-5p(hsa-miR-6832-5p)在膀胱肿瘤组织、膀胱肿瘤细胞中的表达,探讨其对富含脯氨酸11(PRR11)基因表达的干扰作用及对细胞增殖和迁移的影响。方法实时荧光定量聚合酶链反应(qPCR)检测膀胱肿瘤组织、癌旁组织、膀胱肿瘤细胞株和正常膀胱上皮细胞中hsa-miR-6832-5p的表达水平。生物信息学预测并采用双荧光素酶报告基因验证hsa-miR-6832-5p的下游靶基因。分别转染hsa-miR-6832-5p模拟物和miR-NC至hsa-miR-6832-5p表达水平最低的膀胱肿瘤细胞株,命名为miR-6832-5p组和miR-NC组。qPCR检测转染后细胞中hsa-miR-6832-5p和靶基因mRNA的表达水平。Western blot检测靶基因蛋白的表达水平。四甲基偶氮唑蓝(MTT)实验和Transwell实验分别检测细胞增殖和迁移能力。结果膀胱肿瘤组织中hsa-miR-6832-5p的表达低于癌旁组织(P<0.01),膀胱肿瘤细胞株中hsa-miR-6832-5p的表达均低于正常膀胱上皮(P<0.05),其中T24细胞表达水平最低(P<0.01)。生物信息学和双荧光素酶报告基因显示hsa-miR-6832-5p可直接作用于富含脯氨酸11基因(PRR11)的3′-非翻译区(P<0.01)。miR-6832-5p组细胞中hsa-miR-6832-5p的表达量明显高于miR-NC组(P<0.01)。miR-6832-5p组细胞中PRR11的表达量明显低于miR-NC组(P<0.01)。Western blot结果与qPCR结果一致。与miR-NC组比较,转染hsa-miR-6832-5p后膀胱肿瘤细胞增殖能力明显下降(P<0.05),细胞的迁移能力明显下降(P<0.01)。结论 Hsa-miR-6832-5p在膀胱肿瘤组织和细胞株中表达明显降低,hsa-miR-6832-5p可通过下调PRR11基因的表达抑制膀胱肿瘤细胞的增殖和迁移。Objective To analyze the expression of hsa-microRNA-6832-5p(hsa-miR-6832-5p)in bladder tumor tissues and bladder tumor cells,and to explore its interference on the expression of proline-rich protein 11(PRR11)gene in bladder tumor cells and its effect on cell proliferation and migration.Methods Real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression level of hsa-miR-6832-5p in bladder tumor tissues,paratumorous tissues,bladder tumor cell lines and normal bladder epithelial cells.Bioinfbrmatics predicts and the dual luciferase reporter gene validates the downstream target gene of hsa-miR-6832-5p.hsa-miR-6832-5p or miR-NC were transfected to the bladder tumor cell lines with the lowest expression level of hsa-miR-6832-5p,respectively,and named as miR-6832-5p group and miR-NC group.qPCR was used to detect the expression levels of hsa-miR-6832-5p and target gene mRNA in the transfected cells.Western blot was used to detect the expression level of target gene protein.Methyl thiazolyl tetrazolium(MTT)assay and transwell assay were used to detect cell proliferation and migration,respectively.Results The expression of hsa-miR-6832-5p was lower in bladder tumor tissues than in adjacent tissues(P<0.01).The expression of hsa-miR-6832-5p in bladder tumor cells was lower than that in normal bladder epithelial cells(P<0.05),and T24 cells had the lowest hsa-miR-6832-5p expression level(P<0.01).Bioinformatics and dual luciferase reporter genes showed that hsa-miR-6832-5p can directly act on the 3-untranslated region of the PRR11 gene(P<0.01).The expression of hsa-miR-6832~5p in the miR-6832~5p group was significantly higher than that in the miR-NC group(P<0.01).The expression of PRR11 in the miR-6832-5p group was significantly lower than that in the miR-NC group(P<0.01).Western blot results were consistent with qPCR results.Compared with the miR-NC group,the pro­liferation of bladder tumor cells was significantly decreased after transfection with hsa^miR-6832-5p(P<0.05),and the migration ability o

关 键 词：膀胱肿瘤 微RNAS 富含脯氨酸11 肿瘤细胞 培养的 细胞增殖 细胞迁移 

分 类 号：R737.14[医药卫生—肿瘤]