基础研究(Basic Research)

miR-124抑制结肠癌细胞增殖和侵袭与TET蛋白家族的关系

Published at: 2016年第25卷第4期

吴明浩 1 , 张渝 1 , 田力 2 , 唐岸柳 2
1 湖南师范大学第一附属医院/ 湖南省人民医院 消化内科,湖南 长沙 410005
2 中南大学湘雅三医院 消化内科,湖南 长沙 410013
通讯作者 岸柳 唐 Email: tanganliu1983@163.com
DOI: 10.3978/j.issn.1005-6947.10.3978/j.issn.1005-6947.2016.04.009
基金:
国家自然科学基金资助项目 81302076

摘要

目的:探讨miR-124是否通过靶向调节TET蛋白家族的表达而抑制结肠癌细胞增殖与侵袭。方法:用双荧光素酶报告基因检测系统分别检测miR-124对TET家族(TET1、TET2、TET3)的3'UTR-荧光素酶活性的影响;用qRT-PCR与Western blot检测miR-124模拟物转染结肠癌HT29细胞后TET家族的mRNA与蛋白表达水平的变化;用MTS和Transwell实验观察HT29细胞转染miR-124模拟物及TET siRNA后增殖和侵袭能力的变化。结果:双荧光素酶报告基因检测结果显示,各TET mRNA的3'UTR均被miR-124特异性结合,其荧光素酶活性被明显抑制(均P<0.05);转染miR-124模拟物后的HT29细胞TET的mRNA与蛋白表达水平明显减低(均P<0.05);HT29细胞转染miR-124模拟物或TET siRNA后,增殖和侵袭能力均明显降低(均P<0.05)。结论:miR-124可能通过直接靶向调控TET基因的表达,而抑制结肠癌细胞增殖和侵袭。


Relationship between inhibitory effect of miR-124 on proliferation and invasion in colon cancer cells and TET protein family

Abstract

Objective: To investigate whether miR-124 inhibits proliferation and invasion of colon cancer cells by targeted regulation of TET protein family expressions. Methods: The effect of miR-124 on luciferase activity of TET family (TET1, TET2 and TET3) was examined respectively by using a dual-luciferase reporter gene system. The changes in mRNA and protein expressions of TET family in colon cancer HT29 cells after transfection of miR-124 mimics were tested by qRT-PCR and Western blot analysis. The changes in proliferative and invasion abilities of HT29 cells after transfection of miR-124 mimics or TET siRNAs were determined by MTS and Transwell assay, respectively. Results: The results of the dual-luciferase reporter gene assay demonstrated that the 3’UTR of each TET mRNA was specifically matched by miR-124 and their luciferase activities were significantly inhibited (all P<0.05). The mRNA and protein expressions of TET in HT29 cells were significantly down-regulated after miR-124 mimics transfection (all P<0.05). The proliferative and invasion abilities of HT29 cells were significantly reduced after either miR-124 mimics or TET siRNAs transfection (all P<0.05). Conclusion: MiR-124 inhibits proliferation and invasion of colon cancer cells probably through direct regulation of TET expression.


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引用

引用本文: 明浩 吴, 渝 张, 力 田, 岸柳 唐. miR-124抑制结肠癌细胞增殖和侵袭与TET蛋白家族的关系[J]. 中国普通外科杂志, 2016, 25(4): 518-523.
Cite this article as: WU Minghao, ZHANG Yu, TIAN Li, TANG Anliu . Relationship between inhibitory effect of miR-124 on proliferation and invasion in colon cancer cells and TET protein family [J]. Chin J Gen Surg, 2016, 25(4): 518-523.