摘要
目的 :探讨沉默核因子-κB(nuclear factor kappa B,NF-κB)p65对鼻咽癌CNE-2干细胞增殖、凋亡和迁移能力的影响。方法 :将沉默NF-κB p65基因的重组慢病毒感染CNE-2干细胞,应用蛋白质印迹法检测CNE-2干细胞中NF-κB p65、Bcl-2和Bax蛋白的表达,免疫荧光实验观察NF-κB p65在细胞核和细胞质中的表达变化,CCK-8法检测细胞的增殖能力,FCM法检测细胞的凋亡,Transwell小室法检测细胞的迁移能力。结果 :沉默NF-κB p65基因的重组慢病毒感染后,CNE-2干细胞中NF-κB p65和Bcl-2蛋白的表达水平低于空白对照组(CNE-2干细胞未进行任何转染)和阴性对照组(CNE-2干细胞转染阴性对照慢病毒)(P值均<0.01),Bax蛋白的表达水平高于空白对照组和阴性对照组(P值均<0.01);而空白对照组和阴性对照组细胞中NF-κB p65、Bcl-2和Bax蛋白表达水平间的差异无统计学意义(P值均>0.05)。沉默NF-κB p65基因的重组慢病毒感染后,CNE-2干细胞的增殖能力和迁移能力下降(P<0.05和P<0.01),而凋亡率升高(P<0.01)。结论 :NF-κB p65可介导鼻咽癌CNE-2干细胞的增殖、凋亡及迁移能力,其作用可能与调节Bcl-2和Bax的表达有关。
Objective： To investigate the effects of nuclear factor kappa B（NF-κB） p65 gene-silencing on proliferation, apoptosis and migration of human nasopharyngeal carcinoma CNE-2 stem cells.Methods： The recombination lentivirus to silence NF-κB p65 gene was infected into CNE-2 stem cells, then the expression levels of NF-κB p65, Bcl-2 and Bax proteins were detected by Western blotting. The expression of NF-κB p65 in nucleus and cytoplasm was observed by immunofluorescence assay. The abilities of proliferation, apoptosis and migration of CNE-2 stem cells after infection with recombination lentivirus were determined by CCK-8, FCM and Transwell methods, respectively.Results： The expression levels of NF-κB p65 and Bcl-2 proteins in CNE-2 stem cells after infection with recombination lentivirus were lower than those in CNE-2 stem cells without any infection（as a blank control） or after infection with negative control lentivirus（as a negative control）（all P 0.01）, but the expression of Bax protein was higher（both P 0.01）. The expression levels of NF-κB p65, Bcl-2 and Bax proteins between blank control and negative control groups were not significantly different（all P 0.05）. The abilities of proliferation and migration of CNE-2 stem cells after infection with recombination lentivirus were decreased（P 0.05 and P 0.01, respectively）, and the apoptotic rate was increased（P 0.01）.Conclusion： NF-κB p65 can mediate the proliferation, apoptosis and migration of nasopharyngeal carcinoma CNE-2 stem cells by regulating the expressions of Bcl-2 and Bax.
出处
《肿瘤》
CAS
CSCD
北大核心
2016年第2期158-165,共8页
Tumor
基金
贵州省卫生计生委科学技术基金立项课题(编号:gzwjkj2014-2-167)
贵州省科技计划项目[编号:黔科合SY字(2015)3046]~~
关键词
鼻咽肿瘤
肿瘤干细胞
NF-ΚB
细胞增殖
细胞凋亡
细胞迁移分析
Nasopharyngeal neoplasms
Neoplastic stem cells
Nuclear factor kappa B
Cell proliferation
Apoptosis
Cell migration assays
