miR-221与miR-222通过靶基因TIMP3调控胶质瘤细胞侵袭能力  

Modulation of glioma cell invasion by miR-221 and miR-222 through targeting TIMP3

作　　者：翟博智[1] 张春智[2] 韩磊[2] 浦佩玉[2] 康春生[2] 

机构地区：[1]天津环湖医院神经外科,300060 [2]天津医科大学总医院神经外科,天津市神经病学研究所神经肿瘤研究室,教育部"中枢神经系统创伤修复与再生"重点实验室,天津市"神经损伤变异与再生"重点实验室

出　　处：《中华神经外科杂志》2011年第7期701-704,共4页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金资助项目（30772231,30901772）;教育部新世纪优秀人才支持计划资助项目（NCET-07-0615）

摘　　要：目的 探讨miR-221和miR-222在胶质瘤细胞侵袭过程中的作用及其机制.方法 采用反义寡核苷酸下调miR-221和miR-222表达,Transwell、Western blot、荧光素酶实验及体内实验分别检测细胞侵袭能力、体内肿瘤生长、相关基因蛋白表达变化及靶基因鉴定.结果 下调miR-221和miR-222表达能明显抑制胶质瘤细胞侵袭能力,同时相关侵袭蛋白MMP2和MMP9表达下降.miRNA靶基因预测软件分析、Western blot、荧光素酶实验证实TIMP3是miR-221和miR-222的靶基因.裸鼠皮下肿瘤模型显示下调miR-221和miR-222表达抑制体内肿瘤生长.免疫组化发现TIMP3表达上调,MMP2和MMP9表达下调.结论 在胶质瘤细胞中,侵袭相关蛋白TIMP3是miR-221和miR-222的一个新靶基因.Objective To investigate the role and mechanism of miR -221 and miR -222 in glioma cell invasion. Method After reduction of miR -221 and miR -222 by antisense oligonucleotides, cell invasion,invasion related - gene expression and target identification were determined by Transwell assay, Western blot analysis and luciferase reporter assay,respectively. Moreover,the effects of miR -221 and miR -222 on xenograft tumors in nude mice were also observed. Results Down - regulation of miR - 221 and miR - 222 decreased glioma cell invasion in vitro, and inhibited glioma growth in a subcutaneous mouse model. Furthermore,down -regulation of miR -221 and miR - 222 resulted in obvious inactivation of MMP2 and MMP9. Western blot analysis and luciferase reporter assay showed that the reduction of miR - 221 and miR -222 repressed TIMP3 protein expression and TIMP3 mRNA 3＇UTR existed the biding sites of miR -221 and 222. Conclusions TIMP3 is a novel target of miR -221 and miR -222 in glioma cell invasion.

关 键 词：神经胶质瘤 MIR-221 MIR-222 TIMP3 侵袭 

分 类 号：R739.41[医药卫生—肿瘤]