HAUSP表达水平对胶质瘤细胞增殖和凋亡的影响

摘要

Objective To investigate the function and possible mechanisms of HAUSP expression on the proliferation and apoptosis of glioma cells.Methods HAUSP overexpression and low expression plasmids were constructed and transfected U 87 and U251 cell lines by virus vector.Real-time reverse-transcription PCR and Western blotting were utilized to detect mRNA lev-els and protein expression of HAUSP and NANOG in theglioma cells,respectively.The abilities of cellular proliferation and ap-optosis were measured by mosmanntetrazolium test(MTT)assay and flow cytometry(FCM).Results HAUSP overexpression and low expression U87 and U251 cell lines were constructed successfully.In HAUSP overexpression group,we found an obvi-ous increase in the mRNA and protein expression levels of HAUSP compared to the control group.Simultaneously,the mRNA of NANOG was not significantly different,but NANOG protein increased obviously,which increased the proliferation of glioma cells and reduced the apoptosis of those cells.In HAUSP lowexpression group,we found an obvious decrease in the mRNA and protein expression level of HAUSP as compared to the control group.Simultaneously,the mRNA of NANOG was not signifi-cantly different,but NANOG protein decreased obviously,which decreased the proliferation of glioma cells and increased the ap-optosis of those cells.Conclusion HAUSP overexpression can promote the proliferation of glioma cells and increase NANOG protein level.Therefore,HAUSP may maintain the proliferation activity of glioma cells by maintaining deubiquitination of NANOG.%目的 探讨调控疱疹病毒相关泛素特异性蛋白酶(HAUSP)对胶质瘤细胞增殖和凋亡的影响.方法 构建HAUSP过表达及低表达质粒,转染胶质瘤 U87、U251细胞,上调或下调胶质瘤细胞中 HAUSP表达水平.荧光定量PCR、Western blot检测胶质瘤细胞中HAUSP mRNA和蛋白表达水平;利用四甲基偶氮唑盐比色法(MTT)检测细胞增殖水平;流式细胞术检测细胞凋亡变化.同时检测各组细胞NANOG mRNA 和蛋白表达情况.结果 成功构建稳定HAUSP过表达和低表达U87、U251细胞株;与空白对照组相比,HAUSP过表达组细胞中,HAUSP的mRNA、蛋白水平均明显增加,NANOG mRNA水平无明显变化,但NANOG蛋白水平增加,同时细胞增殖能力增强,细胞凋亡水平降低;与空白对照组相比,HAUSP干扰组细胞中,HAUSP的mRNA、蛋白水平表达均降低,NANOG mRNA水平无明显变化,但NANOG蛋白水平降低,同时细胞增殖能力减弱,细胞凋亡水平增加.结论 HAUSP过表达可促进胶质瘤细胞的增殖,这一过程中伴随着NANOG蛋白水平的增高.因此,HAUSP可能通过维持NANOG去泛素化状态而维持胶质瘤细胞的增殖活性.