PEA3和EPHA2在脑胶质母细胞瘤中的表达及在Wnt/β-catenin通路的作用

Expression of PEA3 and EPHA2 in glioblastoma and role in the Wnt/β-catenin pathway

目的探讨多瘤病毒增强活化子3(PEA3)及红细胞生成素产生肝细胞受体2(EPHA2)在脑胶质母细胞瘤中的表达及在Wnt/β-catenin信号通路的作用。方法构建脑胶质瘤U87细胞基因转染模型,将细胞系分为5组:空白组、PEA3干扰组、PEA3干扰空载组、EPHA2干扰组、EPHA2干扰空载组,Western blotting实验检测细胞中EPHA2、PEA3的蛋白表达水平,细胞活力检测(CCK-8)实验检测PEA3、EPHA2对细胞增殖能力的影响,定量反转录聚合酶连锁反应(qRT-PCR)实验检测Wnt/β-catenin通路下游基因转录因子4(TCF-4)、淋巴细胞增强结合因子1(LEF1)表达水平。结果Western blotting实验结果显示:与空白组相比,EPHA2在EPHA2干扰组中表达量降低,PEA3在PEA3干扰组和EPHA2干扰组中表达量降低,差异均具有统计学意义(P<0.05);5组细胞Wnt1、β-catenin蛋白表达量比较,差异无统计学意义(P>0.05)。CCK-8实验结果显示,与空白组相比,PEA3干扰组和EPHA2干扰组的细胞增殖率明显下降,差异均有统计学意义(P<0.05)。qRT-PCR结果显示,TCF-4、LEF1基因在PEA3干扰组、EPHA2干扰组中表达量下降,差异均有统计学意义(P<0.05)。结论通过干扰PEA3和EPHA2基因可降低胶质母细胞瘤的增殖能力,但PEA3和EPHA2是否通过Wnt/β-catenin通路促进胶质母细胞瘤的增殖能力值得进一步研究证实。

Objective To explore the expression of polyomavirus enhancer activator 3(PEA3)and erythropoietin-producting hepatocellular receptor 2(EPHA2)in glioblastoma and their expression in Wnt/β-catenin pathway.Methods A gene transfection model of U87 glioma cells was constructed,and the cell lines were divided into 5 groups:blank group,PEA3 interference group,PEA3 interference empty group,EPHA2 interference group,and EPHA2 interference empty group.The protein expression levels of EPHA2 and PEA3 in U87 cells were detected through Western blotting experiments;The proliferation rate was detected by CCK-8 test,and the the expression levels of T cytokines factor-4(TCF-4)and lymph enhancer factor 1(LEF1)genes,the downstream genes of Wnt/β-catenin pathway,were detected by quantitative real time polymerase chain reaction(qRT-PCR)assay.Results The Western blotting experiment showed that compared with the blank group,the expression level of PEA3 was reduced in the PEA3 interference group and EPHA2 interference group,while the expression level of EPHA2 was reduced in the EPHA2 interference group,and the differences were statistically significant(P<0.05);There was no statistically significant difference in the expression levels of Wnt1 andβ-catenin proteins among 5 groups(P>0.05).The CCK-8 experiment showed that compared with the blank group,the cell proliferation rates of the PEA3 interference group and the EPHA2 interference group were significantly reduced,and the differences were statistically significant(P<0.05).The qRT-PCR results showed that the expression levels of TCF-4 and LEF1 genes decreased in the PEA3 interference group and EPHA2 interference group,and the differences were statistically significant(P<0.05).Conclusion Interference with PEA3 and EPHA2 genes can reduce the proliferation ability of glioblastoma;whether PEA3 and EPHA2 promote the proliferative capacity of glioblastoma through the Wnt/β-catenin pathway deserves further investigation.