缺氧诱导因子通过FOXC1促进脑胶质瘤细胞侵袭  

Hypoxia-inducible factor enhances invasion ability of glioblastoma by regulating FOXC1

作　　者：陈威 彭灿 励勇[1] 章建飞 王新东[1] CHEN Wei;PENG Can;LI Yong;ZHANG Jian-fei;WANG Xin-dong(Department of Neurosurgery,The Affiliated Hospital of Medical School of Ningbo University,Ningbo315211,China;Ningbo Diagnostic Pathology Center,Ningbo 315021,China)

机构地区：[1]宁波大学医学院附属医院神经外科,浙江宁波315211 [2]宁波市临床病理诊断中心,浙江宁波315021

出　　处：《中国病理生理杂志》2021年第7期1219-1226,共8页Chinese Journal of Pathophysiology

基　　金：宁波市科技计划项目(No.2018A610307)。

摘　　要：目的:探究在缺氧情况下,缺氧诱导因子(HIF)能否改变胶质瘤细胞内叉头框蛋白C1(FOXC1)的表达水平,并对胶质瘤细胞的侵袭产生影响。方法:通过RT-qPCR和Western blot实验检测正常氧条件及缺氧条件下胶质瘤细胞系U87和U251中FOXC1的mRNA和蛋白表达水平以及HIF-1α和HIF-2α的蛋白水平变化。缺氧情况下,在胶质瘤U87和U251细胞系中分别转染HIF-1αsiRNA和HIF-2αsiRNA,通过Western blot检测转染HIF-1αsiRNA和HIF-2αsiRNA后U87和U251细胞中FOXC1的蛋白水平变化。体外Transwell实验比较正常氧条件和缺氧情况下U87细胞和U251细胞的侵袭能力,检测转染FOXC1 siRNA后胶质瘤细胞系U87和U251侵袭能力改变及与侵袭能力相关标志蛋白(N-cadherin、E-cadherin和vimentin)的表达水平。结果:缺氧条件下,FOXC1的mRNA和蛋白表达明显升高,HIF-1α和HIF-2α的蛋白水平也明显升高,并随缺氧程度的增加而升高(P<0.05);缺氧情况下,将HIF-1αsiRNA转染至胶质瘤细胞系U87和U251后,发现细胞系中FOXC1的蛋白表达水平较阴性对照组无明显变化;转染HIF-2αsiRNA后,U87细胞系和U251细胞系中FOXC1的蛋白表达水平较阴性对照组明显降低(P<0.05);缺氧条件下,胶质瘤细胞系U87和U251的侵袭能力较正常氧条件下强(P<0.05)。但转染FOXC1 siRNA后,U87和U251细胞的侵袭能力较阴性对照组明显减弱,且侵袭相关标志蛋白N-cadherin和vimentin的表达降低,E-cadherin的表达升高(P<0.05)。结论:缺氧情况下,HIF-2α通过升高FOXC1的表达,进而增强恶性胶质瘤细胞的侵袭能力。AIM:To explore whether hypoxia-inducible factor(HIF)changes the expression level of forkhead box C1(FOXC1)in glioma cells and affects the invasiveness of glioma cells.METHODS:The mRNA and protein ex-pression of FOXC1 and the protein expression of HIF-1αand HIF-2αin glioma cell lines U87 and U251 under normoxic and hypoxic conditions were detected by RT-qPCR and Western blot,and the protein expression of FOXC1 in U87 and U251 cells were detected by Western blot after transfection of HIF-1αsiRNA and HIF-2αsiRNA under hypoxia.In vitro Transwell assay was used to compare the invasive ability of U87 cells and U251 cells under normoxic and hypoxic condi-tions.The changes of invasive ability and the expression of marker proteins(N-cadherin,E-cadherin and vimentin)relat-ed to invasive ability of U87 cells and U251 cells transfected with FOXC1 siRNA were also detected.RESULTS:The mRNA and protein expression of FOXC1 were significantly increased under hypoxia,as well as the protein expression of HIF-1αand HIF-2α,which was increased with the increase in hypoxia degree(P<0.05).After transfection of HIF-1αsiRNA into U87 cells and U251 cells under hypoxia,no significant change in the protein expression of FOXC1 was ob-served as compared with negative control group(P<0.05).After transfection of HIF-2αsiRNA,the protein level of FOXC1 in U87 cells and U251 cells was significantly lower than that in negative control cells,and the invasiveness of U87 cells and U251 cells under hypoxia was increased compare with that under normal oxygen condition(P<0.05).However,after FOXC1 siRNA transfection,the invasive ability of the 2 cell lines was significantly decreased compare with that of the negative control cells,and the expression of invasion-related marker proteins N-cadherin and vimentin was decresed,while the expression of E-cadherin was increased(P<0.05).CONCLUSION:Under hypoxia,HIF-2αincreases the ex-pression of FOXC1,and then enhances the invasion ability of malignant glioma cells.

关 键 词：缺氧诱导因子2α 叉头框蛋白C1 胶质瘤 细胞侵袭 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学] R739.4[医药卫生—基础医学]