慢病毒介导的CIB1过表达对U87胶质瘤细胞WNT信号通路的影响

Effect of CIB1 overexpression mediated by lentivirus on WNT signaling pathway in U87 glioma cells

目的:探讨慢病毒介导的钙整合素结合蛋白1(calcium integrin-binding protein 1,CIB1)在U87胶质瘤细胞中的表达情况及对WNT信号通路的影响。方法:体外培养U87胶质瘤细胞,将携带CIB1基因的LV-CIB1慢病毒转染至U87细胞,获得CIB1稳定过表达的U87脑胶质瘤细胞。根据感染病毒的不同,将细胞分为LV-CIB1过表达组、阴性对照病毒组和正常U87细胞对照组。采用蛋白免疫印迹法检测U87胶质瘤细胞中相关蛋白表达情况,Edu法检测CIB1过表达对U87细胞增殖的影响。结果:LV-CIB1感染U87胶质瘤细胞的最佳感染复数值为4。LV-CIB1过表达组CIB1蛋白表达水平显著高于正常U87细胞对照组和阴性对照病毒组。与正常U87细胞对照组及阴性对照病毒组相比,LV-CIB1过表达组WNT信号通路中关键蛋白GSK-3β表达量降低,β-catenin、cyclin D1及Phospho-GSK-3β(Ser9)的表达水平升高,Phospho-GSK-3β(Tyr216)无明显变化。结论:CIB1过表达促进U87胶质母细胞瘤的生长增殖。WNT信号通路可能是CIB1发挥作用的途径之一,提示可以通过干预WNT信号通路来调节CIB1介导的U87胶质瘤细胞增殖。

Objective:To investigate the effect of calcium integrin-binding protein 1(CIB1)overexpression mediated by lentivirus on WNT signaling pathway in U87 glioma cells.Methods:U87 glioma cells were cultured in vitro,and LV-CIB1 lentiviruses carrying CIB1 gene were transfected into U87 glioma cells,then CIB1 overexpression U87 glioma cells were obtained.According to the different infection viruses,the cells were divided into LV-CIB1 overexpression group,negative control virus group and normal U87 cell group.Western Blot was used to detect the expression of related proteins in U87 glioma cells,and Edu method was used to detect the effect of CIB1 overexpression on the proliferation of U87 glioma cells.Results:The best multiplicity of infection(MOI)was four in LV-CIB1 infected U87 glioma cells.The expression of CIB1 protein in LV-CIB1 overexpression group was significantly higher than that in normal U87 cell group and negative control virus group.Compared with normal U87 cell group and negative control virus group,the expression of GSK-3βin LV-CIB1 overexpression group was decreased,while the expression ofβ-catenin,cyclin D1 and Phospho-GSK-3β(Ser9)were increased,and the expression of Phospho-GSK-3β(Tyr216)did not change significantly.Conclusion:Overexpression of CIB1 promote the proliferation of glioblastoma.WNT signaling pathway may play a role in CIB1,suggesting that the proliferation of U87 glioma cells mediated by CIB1 can be regulated by interfering with WNT signaling pathway.