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过表达miR-18a靶向ATM对结肠癌细胞增殖和迁移的影响

Study on the Effect of Overexpression of miR-18a on Cellular Proliferation and Migration by TargetingATM in Human Colorectal Cancer Cells

作者:刘明佳, 陈利弘, 胡开锋等

Author:LIU Ming-jia, CHEN Li-hong, HU Kai-feng. et al

收稿日期:          年卷(期)页码:2016,47(4):451-457

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:miR-18a ATM 增殖 迁移 结肠癌细胞 HCT116

Key words:miR18a ATM Proliferation Migration Colorectal cancer cells HCT116

基金项目:

中文摘要

目的 研究miR-18a通过靶基因共济失调-毛细血管扩张突变基因 (ataxia telangiectasia mutated gene, ATM)调控结肠癌细胞的生物学特性。方法 通过软件预测miR-18a其中一个靶基因。设计合成miR-18a的模拟剂(mimics)和抑制剂(inhibitor),通过转染不同组分上调或下调内源性miR-18a在结肠癌细胞株HCT116中的表达。采用实时荧光定量(qRT)-PCR、Western blot、 MTT法、克隆形成实验、Transwell等方法,检测过表达miR-18a调控ATM基因的表达对体外结肠癌细胞增殖及迁移能力的影响。结果 软件预测发现miR-18a的其中一个靶基因是ATM。通过瞬时转染,过表达内源性miR-18a,可降低HCT116细胞内靶基因ATM蛋白的表达水平。 转染miR-18a mimics后能够下调HCT116细胞的增殖活力,同时显著降低HCT116细胞的克隆形成能力、横向迁移能力及纵向侵袭能力,以上各项改变均与miR-18a过表达有关。结论 证实了miR-18a通过负向调控ATM的表达,抑制结肠癌细胞HCT116的增殖和迁移能力。

英文摘要

Objective To study the regulation to colon cancer cellular biological properties through miR-18a targeting ataxia-telangiectasia mutated gene (ATM). Methods A target of miR-18a was predicted by using bioinformatics tools. The miR-18a mimics and inhibitors were designed and synthesized. The expression of endogenous miR-18a in colon cancer cell line HCT116 was up-regulated or down-regulated by transfection. The effect of overexpression of miR-18a on cellular proliferation, invasion and migration via regulation of ATM gene expression was confirmed in vitro by using qRT-PCR, Western blot, MTT assay, clone forming assay and Transwell method, respectively. Results ATM was identified as a potential target gene of miR-18a in the bioinformatics analysis. In addition, through transient transfection leading to the overexpression of miR-18a in HCT116 cell, the expression level of ATM was decreased. Down-regulation of HCT116 cell proliferation activity while significantly reducing HCT116 cell clone forming ability, lateral migration ability and longitudinal invasion ability were observed after transfected with miR-18a mimics. All of the changes were related to the overexpression of miR-18a. Conclusion miR-18a inhibited the proliferation and migration of colon cance cell HCT116 through negative regulation of ATM expression.

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