GOLPH3通过WNT信号通路调控胶质瘤细胞增殖及其机制的研究

GOLPH3 regulates the proliferation of glioma cells by WNT signaling pathway and the underlying mechanism

目的探讨高尔基体磷蛋白3(GOLPH3)、WNT/β-catenin信号通路对胶质瘤细胞增殖的影响及机制。方法采用脂质体转染法将siRNA转染至胶质瘤U251细胞中,将转染negative control siRNA oligo和各siRNA oligo的细胞分别设为阴性对照组(siNC组)和下调组(siGOLPH3组、siWNT2组、siβ-catenin组)。采用蛋白质印迹法(Western blot)检测相关蛋白表达水平,EdU法检测对胶质瘤细胞增殖的影响。结果siGOLPH3组细胞中WNT2蛋白相对表达量低于siNC组,siWNT2组细胞中β-catenin蛋白相对表达量低于siNC组,差异均有统计学意义(P﹤0.05)。siWNT2组细胞中GOLPH3蛋白相对表达量与siNC组比较,差异无统计学意义(P﹥0.05)。与siNC组相比,siGOLPH3组、siWNT2组、siβ-catenin组细胞增殖率均降低,差异均有统计学意义(P﹤0.05)。结论下调GOLPH3后可以降低WNT2的蛋白表达水平,下调WNT2后可以降低其下游β-catenin蛋白的表达水平,下调GOLPH3、WNT2、β-catenin均能抑制胶质瘤细胞的增殖。GOLPH3可能通过WNT/β-catenin信号通路影响胶质瘤细胞的增殖。

Objective To investigate the effect of Golgi phosphoprotein 3(GOLPH3)and WNT/β-catenin signaling pathway on the cells proliferation in glioma,and to clarify the underlying mechanism.Method The siRNA oligos were transfected into glioma U251 cells,cells that were transfected with negative control siRNA oligo or siRNA oligo were set as negative control group(siNC group)and down-regulation group(siGOLPH3 group,siWNT2 group and siβ-catenin group),respectively.Protein expression level were explored by Western blot,and EdU assay was employed to determine the cell proliferation.Result In siGOLPH3 group,the WNT2 protein relative expression were lower compared to siNC group,besides,theβ-catenin protein relative expression in siWNT2 group were also lower than that in siNC group,the differences were statistically significant(P<0.05).No evident difference regarding the relative expression of GOLPH3 protein in siWNT2 group and siNC group were noted(P>0.05).Compared with the siNC group,the cell proliferation rate of siGOLPH3 group,siWNT2 group,and siβ-catenin group were all decreased significantly,demonstrating statistical significance(P<0.05).Conclusion Down-regulation of GOLPH3 could reduce the expression of WNT2 protein level in glioma cells,and then subsequently decrease the expression of downstreamβ-catenin,meanwhile,downregulation of GOLPH3,WNT2 andβ-catenin can inhibit the proliferation of glioma cells.GOLPH3 regulates glioma cells proliferation probably through the WNT/β-catenin signaling pathways.