miRNA-20a在膀胱癌中的表达及其机制研究

发布时间：2018-09-08 11:54

【摘要】：目的:探讨miRNA-20a在膀胱癌组织中的表达及其机制。方法:收集2014年1月至2015年1月昆明医科大学第二附属医院96例患者的组织标本,运用实时定量聚合酶链反应(q RT-PCR)方法检测miRNA-20a在膀胱癌组织和癌旁组织中的表达;生物信息学方法预测miRNA-20a的靶基因并采用双荧光素酶报告基因实验进行验证;q RT-PCR、Western blot以及细胞免疫荧光分别检测人膀胱癌细胞系T24和J82细胞转染miRNA-20a模拟物或阴性对照后对靶基因mRNA和蛋白表达的影响;CCK-8、Transwell侵袭小室和划痕实验检测过表达miRNA-20a后T24细胞体外增殖、迁移和侵袭能力的改变。结果:miRNA-20a在膀胱癌组织中高表达,且与肿瘤的病理分级、临床分期、转移以及复发密切相关(P0.001);双荧光素酶报告基因实验证实miRNA-20a与人源性长寿保障基因2(Homo sapiens longevity assurance homologue 2,LASS2)的3'-UTR直接靶向结合;转染miRNA-20a模拟物可显著下调膀胱癌细胞中LASS2 m RNA和蛋白的表达(P0.01),增强膀胱癌细胞的增殖、侵袭和迁移能力(P0.01)。结论:miRNA-20a在膀胱癌组织中高表达,且miRNA-20a可通过靶向抑制LASS2促进膀胱癌细胞的增殖、侵袭和迁移,与膀胱癌的发生发展相关。
[Abstract]:Objective: to investigate the expression and mechanism of miRNA-20a in bladder cancer. Methods: from January 2014 to January 2015, 96 tissue specimens from the second affiliated Hospital of Kunming Medical University were collected and the expression of miRNA-20a in bladder cancer and adjacent tissues was detected by real-time quantitative polymerase chain reaction (Q RT-PCR). Bioinformatics method was used to predict the target gene of miRNA-20a, and double luciferase reporter gene assay was used to verify the expression of miRNA-20a mimics or negative control in human bladder cancer cell lines T24 and J82, respectively, by using double luciferase reporter gene assay and immunofluorescence assay. The expression of target gene mRNA and protein was affected by CCK-8 transwell invasion chamber and scratch assay was used to detect the proliferation of T24 cells after overexpression of miRNA-20a in vitro. Changes in migration and invasiveness. Results the expression of 10 miRNA-20a was highly expressed in bladder cancer tissues, and was correlated with the pathological grade and clinical stage of the tumor. There was a close correlation between metastasis and recurrence (P0.001), double luciferase reporter gene experiment confirmed that miRNA-20a was directly targeted to the 3'-UTR of human longevity guarantee gene 2 (Homo sapiens longevity assurance homologue 2 / LASS2. Transfection of miRNA-20a mimics could significantly down-regulate the expression of LASS2 m RNA and protein in bladder cancer cells (P0.01), and enhance the proliferation, invasion and migration of bladder cancer cells (P0.01). ConclusionMmiRNA-20a is highly expressed in bladder cancer tissues, and miRNA-20a can promote the proliferation, invasion and migration of bladder cancer cells by targeting the inhibition of LASS2, which is related to the occurrence and development of bladder cancer.
【作者单位】：昆明医科大学第二附属医院泌尿外科云南省泌尿外科研究所;云南省楚雄州人民医院新区肾脏内科;
【基金】：国家自然科学基金项目(编号:81460384) 云南省科技计划项目(编号:2015FB196) 云南省教育厅科学研究基金项目(编号:2014Z072)资助~~
【分类号】：R737.14

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