文章摘要

miR-124抑制结肠癌细胞增殖和侵袭与TET蛋白家族的关系

作者: 1吴明浩, 1张渝, 2田力, 2唐岸柳
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: 唐岸柳 Email: tanganliu1983@163.com
DOI: 10.3978/.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。
关键词： 结肠肿瘤 微RNAs DNA甲基化 细胞增殖 肿瘤侵润

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

Authors: 1WU Minghao, 1ZHANG Yu, 2TIAN Li, 2TANG Anliu
1 Department of Gastroenterology, Hunan Provincial People’s Hospital/the First Affiliated Hospital, Hunan Normal University, Changsha, 410005, China
2 Department of Gastroenterology, the Third Xiangya Hospital, Central South University, Changsha, 410013, China

CorrespondingAuthor:TANG Anliu Email: tanganliu1983@163.com

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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