miR-520a调控RAB22A表达并抑制结肠癌细胞的增殖和侵袭  

miR-520a inhibits cells proliferation and invasion through regulating RAB22A expression in colon cancer

作　　者：张灵敏[1] 崔曼莉 杜召召 路宁 闫蓉[2] 张明鑫 Zhang Lingmin;Cui Manli;Du Zhaozhao;Lu Ning;Yan Rong;Zhang Mingxin(Department of Anesthesiology,the First Affiliated Hospital of Xi'an Jiaotong University,Shaanxi Xi'an 710061,China;Department of Gastroenterology,the First Affiliated Hospital of Xi'an Medical University,Shaanxi Xi'an 710077,China)

机构地区：[1]西安交通大学第一附属医院麻醉科,陕西西安710061 [2]西安医学院第一附属医院消化内科,陕西西安710077

出　　处：《现代肿瘤医学》2019年第22期3949-3954,共6页Journal of Modern Oncology

基　　金：陕西省重点研发计划项目(编号:2017SF-165);陕西省自然科学基础研究计划一般项目(面上)(编号:2018JM7090)

摘　　要：目的:探讨miR-520a对结肠癌生物学行为的影响及其对RAB22A表达的调控。方法:应用分子生物学技术构建miR-520a的表达质粒(miR-520a)及抑制剂(anti-miR-520a),应用MTT和Transwell探讨miR-520a对结肠癌细胞株增殖和侵袭的影响。实时荧光定量PCR和蛋白印迹检测转染miR-520a对RAB22A的mRNA和蛋白表达水平的影响,构建RAB22A的野生型及突变型3’-UTR报告基因,报告基因分析miR-520a对RAB22A的调控。结果:miR-520a可以显著抑制结肠癌细胞的增殖和侵袭。蛋白印迹显示miR-520a可以显著抑制RAB22A的蛋白表达水平,而对mRNA表达无显著影响。报告基因分析显示miR-520a能够下调RAB22A的野生型3’-UTR报告基因的荧光表达,而对突变型报告基因的荧光表达影响不大。结论:miR-520a可能通过转录后调控癌基因RAB22A的表达抑制结肠癌细胞的增殖和侵袭,在结肠癌中发挥抑癌基因功能。Objective:To investigate the effect of miR-520 a on behavior of colon cancer and its post-transcriptional regulation of RAB22 A expression.Methods:After construction of miR-520 a expression vector(miR-520 a) and miR-520 a inhibitor(anti-miR-520 a),the effect of miR-520 a on proliferation and invasion of colon cancer cell line SW480 was detected by MTT and Transwell respectively.Regulation of miR-520 a on expression of RAB22 A was investigated by real time PCR,Western blot and luciferase reporter assay system.Results:miR-520 a inhibited proliferation and invasion in SW480.Moreover,real time PCR and Western blot revealed that miR-520 a down-regulated the expression of RAB22 A protein,but had no effect on RAB22 A mRNA expression in vitro.Luciferase reporter assay confirmed that miR-520 a regulated RAB22 A expression through certain sequence in 3’-UTR.Conclusion:miR-520 a inhibits proliferation and invasion possibly through post-transcriptionally regulating expression of RAB22 A oncogene,suggesting miR-520 a functions as a tumor suppressor in colon cancer.

关 键 词：miR-520a RAB22A 结肠癌 增殖 侵袭 

分 类 号：R735.3[医药卫生—肿瘤]