贵州医科大学学报

2018, v.43;No.219(12) 1420-1424+1435

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miR-133b靶向下调EGFR对鼻咽癌SUNE-1细胞增殖、迁移及侵袭的影响
Inhibitory Effect of miR-133b on Down-regulation of EGFR Expression in Nasopharyngeal Carcinoma SUNE-1 Cells

闫勇;吴曙辉;朱静静;姜婷婷;
YAN Yong;WU Shuhui;ZHU Jingjing;JIANG Tingting;Department of Otolaryngology,Shanghai Baoshan Integrated Hospital of Traditional Chinese and Western Medicine;

摘要(Abstract):

目的:研究miR-133b对鼻咽癌SUNE-1细胞增殖、迁移及侵袭的影响,并探讨其作用机制。方法:采用q PCR检测人鼻咽癌细胞株CNE-1、SUNE-1、C666-1及正常人鼻咽上皮细胞株NP69中miR-133b及表皮生长因子受体(EGFR)的表达差异,使用荧光光素酶报告基因及蛋白免疫印迹实验测定miR-133b和EGFR在鼻咽癌SUNE-1细胞株中的调控关系;将miR-133b mimics转染至SUNE-1细胞中构建miR-133b过表达模型,同时将EGFR siRNA转染至SUNE-1细胞中构建EGFR低表达模型,通过MTT实验检测细胞的增殖能力、Transwell实验检测细胞的迁移及侵袭能力。结果:与NP69细胞株比较,miR-133b在CNE-1、SUNE-1、C666-1细胞株中低表达(P <0. 05)、且在SUNE-1细胞株中表达最低,而EGFR在在CNE-1、SUNE-1、C666-1中高表达(P <0. 05)、且在SUNE-1细胞株中表达最高;双荧光素酶报告基因及蛋白免疫印迹实验结果显示增强miR-133b的表达后,SUNE-1细胞株中EGFR的表达相应下调(P <0. 05);增强miR-133b的表达或抑制EGFR的表达,鼻咽癌细胞的增殖、侵袭、迁移能力显著减弱(P <0. 05)。结论:miR-133b可能通过负向调控EGFR的表达,从而抑制鼻咽癌细胞的增殖、迁移及侵袭能力。
Objective: To investigate the effect of miR-133 b on the proliferation,migration and invasion of nasopharyngeal carcinoma SUNE-1 cells and discuss the mechanism of its action. Methods:The expression of miR-133 b and epidermal growth factor receptor( EGFR) in human nasopharyngeal carcinoma cell line CNE-1,SUNE-1,C666-1 and normal nasopharyngeal epithelial cell line NP69 were detected by qPCR. The regulatory relationship between miR-133 b and EGFR in nasopharyngeal carcinoma SUNE-1 cell line was detected by fluorescein enzyme reporter gene and Western blot assay. The miR-133 b mimics was transfected into SUNE-1 cells to structure miR-133 b overexpression model,and EGFR siRNA was transfected in SUNE-1 cells to structure EGFR low expression model. Cell proliferation was detected by MTT assay,and cell migration and invasion was detected by Transwell assay.Results: Low expression of miR-133 b in CNE-1,SUNE-1 and C666-1 cell lines was compared with NP69 cell line( P < 0. 05),and the lowest expression was found in SUNE-1 cell lines. In high expression of EGFR in CNE-1,SUNE-1,C666-1( P < 0. 05),the highest expression was found in SUNE-1cell lines. The results of double luciferase reporter gene and Western blotting showed that the expression of EGFR in SUNE-1 cells was down-regulated after the expression of miR-133 b was enhanced( P< 0. 05). Enhancing the expression of miR-133 b or inhibiting the expression of EGFR,the proliferation,invasion and migration of nasopharyngeal carcinoma cells were significantly decreased( P <0. 05). Conclusion: MiR-133 b may inhibit the proliferation,migration and invasion of nasopharyngeal carcinoma cells by negatively regulating the expression of EGFR.

关键词(KeyWords): miR-133b;鼻咽癌;表皮生长因子受体;增殖;迁移;侵袭
miR-133b;nasopharyngeal carcinoma;epidermal growth factor receptor;proliferation;migration;invasion

Abstract:

Keywords:

基金项目(Foundation): 上海市卫生和计划生育委员会科研课题(16PJ057)

作者(Author): 闫勇;吴曙辉;朱静静;姜婷婷;
YAN Yong;WU Shuhui;ZHU Jingjing;JIANG Tingting;Department of Otolaryngology,Shanghai Baoshan Integrated Hospital of Traditional Chinese and Western Medicine;

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DOI: 10.19367/j.cnki.1000-2707.2018.12.011

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