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miR-221与miR-222通过靶基因TIMP3调控胶质瘤细胞侵袭能力
引用本文:翟博智,张春智,韩磊,浦佩玉,康春生. miR-221与miR-222通过靶基因TIMP3调控胶质瘤细胞侵袭能力[J]. 中华神经外科杂志, 2011, 27(7). DOI: 10.3760/cma.j.issn.1001-2346.2011.07.019
作者姓名:翟博智  张春智  韩磊  浦佩玉  康春生
作者单位: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
基金项目:国家自然科学基金资助项目,教育部新世纪优秀人才支持计划资助项目
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.
Abstract:
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  侵袭

Modulation of glioma cell invasion by miR-221 and miR-222 through targeting TIMP3
ZHAI Bozhi,ZHANG Chun-zhi,HAN Lei,PU Pei-yu,KANG Chun-sheng. Modulation of glioma cell invasion by miR-221 and miR-222 through targeting TIMP3[J]. Chinese Journal of Neurosurgery, 2011, 27(7). DOI: 10.3760/cma.j.issn.1001-2346.2011.07.019
Authors:ZHAI Bozhi  ZHANG Chun-zhi  HAN Lei  PU Pei-yu  KANG Chun-sheng
Abstract:
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.
Keywords:miR-221  miR-222  TIMP3
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