Netrin-4对胶质母细胞瘤细胞ERK/MAPK信号通路的影响

目的:通过免疫印迹方法检测不同浓度的Netrin-4(NTN4)重组蛋白对GBM细胞中磷酸化ERK水平的影响,初步探讨NTN4对GBM细胞ERK/MAPK信号通路的调控作用.方法:体外培养野生型GBM细胞:U251 MG细胞及U87MG细胞,加入两种不同浓度的NTN4重组蛋白(5ng/ml,50ng/ml),裂解细胞提取目的蛋白,在不同时间点(0,10min/20min,30min,1h,2h,3h)通过免疫印迹方法检测磷酸化ERK水平变化情况.结果:加入NTN4后,U251 MG细胞和U87MG细胞中的磷酸化ERK水平都有所增加;在U251 MG细胞中,5ng/ml的NTN4作用后1h,磷酸化ERK水平最高,50ng/ml的NTN4作用后10分钟,磷酸化ERK水平最高;在U87MG细胞中,不同浓度的NTN4都在作用10分钟时,磷酸化ERK水平最高.结论:Netrin-4激活胶质母细胞瘤细胞内ERK/MAPK信号通路,是其抑制GBM细胞衰老的可能机制.

Influence from Netrin-4 to ERK/MAPK signal pathway in GBM cell lines

Objective:To investigate the effect of different concentrations of Netrin-4 on the phosphorylation of ERK in GBM cells were detected by Western blot,and the effect of NTN4 on ERK/MAPK signaling pathway in GBM cells were discussed.Methods:U251MG cells and U87MG cells were cultured in vitro.The target protein was extracted from the cells,and the levels of phosphorylated ERK were detected by Western blot (50ng/ml-0,10min/20min,30min,1 h,2h,3h;5ng/ml-0,10min/20min,30min,1 h,2h,3h).Results:After the addition of U251MG,the phosphorylation of ERK in NTN4 cells and U87MG cells increased,and in U251 MG cells,the level of phosphorylated at 1 h was the highest after added 5ng/ml NTN4.The level of phosphorylated ERK was highest at 10min after added 50ng/ml NTN4 in U251MG cells.Meanwhile,the level of phosphorylated ERK was the highest at 10min after added either concentration in U87MG cells.Conclusion:Netrin-4 activates the ERK/MAPK signaling pathway in glioblastoma cell lines,probably which protects GBM cell senescence clarifies the mechanism.