慢病毒介导shRNA靶向ZNF217抑制胶质瘤细胞生长、迁移和侵袭  

Lentivirus-mediated sh RNA targeting ZNF217 suppresses cell growth, migration, and invasion of glioma cells in vitro

作　　者：罗起胜[1] 黄海能[1] 邓元央[1] 黄华东[1] 符黄德[1] 罗琨祥[1] 李传玉[1] 覃成箭[1] 韦展亮[1] 栗学玉[1] 

机构地区：[1]右江民族医学院附属医院神经外科,广西百色533000

出　　处：《南方医科大学学报》2015年第7期1024-1027,1033,共5页Journal of Southern Medical University

基　　金：广西自然科学基金(2014GXNSFBA118143);广西高校科学技术研究项目(YB2014305)

摘　　要：目的探讨癌基因ZNF217对胶质瘤U251细胞迁移、侵袭以及增值的影响以及相应的分子机制。方法构建shRNAZNF217慢病毒干扰载体,在将其包装成成熟的慢病毒后感染胶质瘤U251细胞。Western blot检测ZNF217干扰效率。利用transwell、Boyden、MTT和流式细胞仪分别检测细胞迁移、侵袭、增殖以及细胞周期能力的改变。western blot检测相应基因表达改变。结果与对照细胞相比,ZNF217基因在慢病毒shRNA-ZNF217作用下,其表达水平在胶质瘤U251细胞中显著下调。Transwell和Boyden结果显示,在抑制ZNF217表达后细胞迁移和侵袭能力也明显下降。此外,MTT和流式细胞仪分析结果表明,细胞的增值和周期转化能力也明显降低。机制分析显示,在抑制ZNF271表达后,磷酸化的PI3K/AKT以及癌基因C-Myc和间质标志物N-Cadherin活性减低,而上皮标志物E-Cadherin活性增高。结论 ZNF217在胶质瘤中通过激活PI3K/AKT上调CMyc基因以及调控上皮向间质转化相关基因(Epithelial–mesenchymal transition EMT)从而促进细胞生长、迁移和侵袭。Objective To explore the role of ZNF217 in regulating cell proliferation, migration and invasion in glioma cells. Methods A lentivirus-mediated shRNA-ZNF217 vector was infected into glioma U251 cells, and the interference efficiency was examined by Western blotting. MTT assay, flow cytometry, Transwell assay, and Boyden chamber assay were used to analyze the changes in cell proliferation, migration and invasion. Western blotting was used to detect the changes in ZNF217-related genes in the cells. Results shRNA-ZNF217 transfection significantly inhibited the expression of ZNF217 in U251 cells and suppressed the cell migration, invasion, growth, and cell cycle transition. ZNF217 knockdown downregulated the expression of pPI3, pAKT, C-Myc, and the mesenchyme biomarker N-cadherin, and stimulated the expression of the epithelium biomarker E-cadherin. Conclusion ZNF217 promotes cell migration, invasion, and growth by activating PI3K/AKT signal to upregulate C-Myc and by modulating the genes associated with epithelial-mesenchymal transition in glioma cells.

关 键 词：ZNF217 胶质瘤 SHRNA 

分 类 号：R739.41[医药卫生—肿瘤]