中国癌症杂志 ›› 2014, Vol. 24 ›› Issue (6): 401-405.doi: 10.3969/j.issn.1007-3969.2014.06.001

• 论著 • 上一篇    下一篇

miR-22通过靶向MTDH抑制胶质瘤细胞的生长

李荣国,王剑,杨少陵   

  1. 南华大学附属第一医院急诊科,湖南 衡阳 421001
  • 出版日期:2014-06-30 发布日期:2014-07-29
  • 通信作者: 李荣国 E-mail:nhdxtgh@163.com
  • 基金资助:
    国家自然科学基金(No:31100936)

miR-22 inhibited glioma cells proliferation by targeting MTDH

LI Rong-guo, WANG Jian, YANG Shao-ling   

  1. Emergency Department of the First Affiliated Hospital of University of South China, Hengyang Hunan 421001, China
  • Published:2014-06-30 Online:2014-07-29
  • Contact: LI Rong-guo E-mail: nhdxtgh@163.com

摘要: 背景与目的:miR-22在胃癌、肺癌、结肠癌以及乳腺癌中表达下调,然而其在胶质瘤中的表达情况尚未明确。本研究旨在探讨miR-22是否通过靶向调控MTDH表达抑制胶质瘤细胞生长,从而进一步揭示miR-22的抑瘤机制方法:运用实时定量PCR(quantitative real-time polymerase chain reaction,qRT-PCR)检测75例胶质瘤及17例正常大脑组织中miR-22的表达改变以及与胶质瘤患者预后关系;构建MTDH 3’UTR-荧光素酶报告载体,通过荧光素酶报告检测观察miR-22对MTDH 3’UTR-荧光素酶活性的影响;将miR-22 mimics转染胶质瘤细胞U251,以MTDH siRNA为阳性对照,采用蛋白质印迹法(Western blot)检测其对MTDH蛋白表达水平的影响;然后采用MTT法检测高表达miR-22对U251细胞生长的影响结果:qRT-PCR检测结果显示,miR-22在75例胶质瘤组织中表达下调;Kaplan-Meier生存分析发现,miR-22表达越高,患者生存率越高(P<0.05);在双荧光素酶报告检测显示,miR-22能特异性地与MTDH 3’UTR结合,抑制其荧光素酶活性。Western blot检测结果显示,miR-22过表达或干扰MTDH可抑制MTDH蛋白的表达。MTT检测发现,miR-22过表达或干扰MTDH可抑制人U251细胞的生长,差异有统计学意义(P<0.05)结论:miR-22通过靶向调控MTDH的表达而抑制胶质瘤细胞的生长。

关键词: miR-22, 胶质瘤, 异粘蛋白, 生长

Abstract:

Background and purpose: miR-22 has been reported to be down-regulated in gastric cancer, lung cancer, colorectal cancer, and breast cancer. However, its expression in glioma was still poorly known. This study aimed to explicit whether miR-22 suppresses cell proliferation by targeting MTDH, thus to reveal molecular mechanism that miR-22 functions as a tumor suppressor in glioma. Methods: Quantitative real-time polymerase chain reaction (qRTPCR) was conducted for detecting the expression of miR-22 in gliomas and normal brain tissues. MTDH 3’UTRluciferase vector was constructed and dual-luciferase reporter gene assay was employed to examine the effect of miR-22 on luciferase activity. U251 cells were transfected with miR-22 mimics, and MTDH siRNA as for postive control, then Western blot was performed to detect the expressions of MTDH protein. The proliferation ability of U251 cells was evaluated by MTT assay. Results: miR-22 was down-regulated in glioma tissues. Glioma patients with relatively high expression of miR-22 showed lower mortality compared with low expression of miR-22 by using Kaplan-Meier survival curves. We demonstrated miR-22 could bind to the 3’ untranslated region (UTR) of MTDH and inhibited the luciferase activity. Western blot showed that the expression of MTDH protein was inhibited by restored miR-22 or siR MTDH in U251 cells. Overexpression of miR-22 or siR MTDH inhibited the proliferation of U251 cells. Conclusion: miR-22 suppresses cell proliferation by targeting MTDH in glioma.

Key words: miR-22, Glioma, MTDH, Growth