干扰内皮素-1可抑制鼻咽癌细胞增殖、迁移和侵袭  

Silencing of endothelin-1 suppresses growth,migration,and invasion of nasopharyngeal carcinoma cells in vitro

作　　者：林少雄[1] 张永[2] 鲁娟[3] 刘雄[3] 李湘平[3] 

机构地区：[1]汕头大学医学院第一附属医院耳鼻咽喉科,广州汕头515041 [2]南方医科大学南方医院医务处,广东广州510515 [3]南方医科大学南方医院耳鼻咽喉头颈外科,广东广州510515

出　　处：《南方医科大学学报》2016年第7期915-920,共6页Journal of Southern Medical University

基　　金：国家自然科学基金(81301849)~~

摘　　要：目的探讨干扰内皮素-1(ET-1)基因对鼻咽癌细胞增殖、迁移及侵袭的影响以及相应的分子机制。方法构建ET-1sh RNA慢病毒干扰载体,成功包装慢病毒后感染鼻咽癌5-8F细胞。Western blotting检测ET-1干扰效率。利用MTT、细胞周期检测、平板克隆形成实验、Transwell小室、Boyden小室以及裸鼠皮下成瘤实验分别检测细胞增殖、迁移、侵袭能力的改变。Western blotting检测上皮间质转化(EMT)相应基因表达改变。结果与对照组细胞相比,鼻咽癌细胞5-8F感染慢病毒sh RNAET-1后ET-1的表达水平显著降低。MTT实验结果显示,在抑制ET-1表达后细胞增殖能力显著下降。流式细胞仪检测细胞周期显示干扰ET-1表达,G1细胞数明显增多。平板克隆形成实验结果显示干扰ET-1表达,癌细胞克隆形成能力显著下降。皮下裸鼠成瘤实验结果同样显示,ET-1表达抑制后癌细胞成瘤能力显著下降。Transwell和Boyden小室结果表明,ET-1表达沉默后可显著降低细胞的迁移和侵袭能力。通过检测ET-1沉默后EMT相关分子的表达改变,结果显示上皮标志物E-cadherin和CK18表达增高,而间质标志物Vimentin和N-cadherin表达降低。结论 ET-1在鼻咽癌中可通过调控上皮间质转化相关基因促进细胞增殖、迁移和侵袭能力。Objective To explore the role of endothelin-1（ET-1） gene in regulating the proliferation,migration and invasion of nasopharyngeal carcinoma cells.Methods A lentivirus-mediated sh RNA-ET-1 vector was infected into 5-8F cells,and the interference efficiency was examined with Western blotting.MTT assay,cell cycle analysis,plate colony formation assay,Transwell assay,Boyden chamber assay and tumor growth assay were carried out to analyze the changes in cell proliferation,migration and invasion.The expressions of genes related with epithelial-mesenchymal transition（EMT） were examined using Western blotting.Results sh RNA-ET-1 transfection significantly inhibited the expression of ET-1,and suppressed the growth,migration and invasion of 5-8F cells.ET-1 knockdown enhanced the expression of E-cadherin and CK18 and inhibited the expression of N-cadherin and vimentin.Conclusion ET-1 promotes cell growth,migration and invasion by modulating the genes associated with epithelial-mesenchymal transition in nasopharyngeal carcinoma cells.

关 键 词：内皮素-1 鼻咽癌 迁移 侵袭 

分 类 号：R739.63[医药卫生—肿瘤]