摘要
目的探讨FOXC2过表达对胶质瘤U87细胞增殖、侵袭、迁移的影响及其机制。方法体外培养人脑胶质瘤U87细胞和人脑胶质细胞HEB。U87细胞随机分为空白组(未转染任何质粒)、空载体组(转染pcDNA3.1空载体质粒)、FOXC2过表达组(转染pcDNA3.1-FOXC2过表达质粒)、FOXC2+抑制剂组[转染pcDNA3.1-FOXC2质粒+Wnt/β-catenin信号通路抑制剂FH535(20μmol/L)]。采用qRT-PCR和免疫印迹法检测mRNA和蛋白表达;CCK-8法检测细胞增殖活性,克隆形成实验分析细胞克隆形成能力;Transwell小室实验检测细胞侵袭和迁移能力。结果与HEB细胞比较,U87细胞FOXC2 mRNA表达水平显著升高(P<0.05)。FOXC2过表达,显著促进U87细胞增殖、侵袭、迁移(P<0.05),显著增加Vimentin、Wnt1、β-catenin蛋白表达水平以及细胞克隆形成率(P<0.05),显著降低E-cadherin蛋白表达水平(P<0.05)。抑制Wnt/β-catenin信号通路,显著抑制FOXC2过表达对U87细胞的作用(P<0.05)。结论FOXC2过表达可通过激活Wnt/β-catenin信号通路,促进U87细胞发生上皮间质转化,从而增加胶质瘤U87细胞增殖、侵袭、迁移能力。
Objective To investigate the effect of FOXC2 overexpression on the proliferation,invasion and migration of glioma U87 cells.Methods The human glial cells HEB and glioma U87 cells were cultured in vitro.The U87 cells were randomly divided into four groups,i.e.,blank group(without transfection of plasmid),FOXC2-NC group(transfection of pcDAN3.1-FOXC2 negative plasmid),FOXC2-overexpression group(transfection of pcDNA3.1-FOXC2 mimics),inhibition group(transfection of pcDNA3.1-FOXC2 mimics+inhibitor of Wnt/β-catenin signaling pathway).The mRNA and protein expression levels were detected by qRT-PCR and western blotting,respectively.CCK-8 method,clone formation experiment,and Transwell cell experiment were used to detect cell proliferation,invasion,and migration abilities.Results Compared with HEB cells,the mRNA expression level of FOXC2 in U87 cells significantly increased(P<0.05).FOXC2 overexpression significantly promoted the abilities of U87 cell proliferation,invasion and migration(P<0.05),significantly increased the protein expression level of Vimentin,Wnt1,andβ-catenin,and the cell clone formation rate(P<0.05),and significantly reduced E-cadherin protein expression levels(P<0.05).Inhibition of the Wnt/β-catenin signaling pathway significantly inhibited the effect of FOXC2 overexpression on the U87 cells(P<0.05).Conclusions Overexpression of FOXC2 can promote epithelialmesenchymal transition in the glioma U87 cells through the activation of Wnt/β-catenin signaling pathway,thereby increasing the proliferation,invasion and migration abilities of glioma U87 cells.
作者
潘轲
向春晖
周龙
汪逵
王国堰
PAN Ke;XIANG Chun-hui;ZHOU Long;WANG Wei;WANG Guo-zhen(Department of Neurosurgery,Enshi Tujia And Miao National Center Hospital,Enshi 445000,China)
出处
《中国临床神经外科杂志》
2021年第4期266-269,共4页
Chinese Journal of Clinical Neurosurgery
