Netrin-4对胶质母细胞瘤细胞ERK/MAPK信号通路的影响

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

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:U251MG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,U251MG细胞和U87MG细胞中的磷酸化ERK水平都有所增加;在U251MG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。

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： U251 MG cells and U87 MG cells were cultured in vitro. The target protein was extracted from the cells,and the levels of phosphorylated ERK were detected by Western blot（ 50 ng / ml-0,10 min /20 min,30min,1h,2h,3h; 5ng / ml-0,10 min /20 min,30min,1h,2h,3h）. Results： After the addition of U251 MG,the phosphorylation of ERK in NTN4 cells and U87 MG cells increased,and in U251 MG cells,the level of phosphorylated at 1h was the highest after added 5ng / ml NTN4. The level of phosphorylated ERK was highest at 10 min after added 50 ng /ml NTN4 in U251 MG cells. Meanwhile,the level of phosphorylated ERK was the highest at 10 min after added either concentration in U87 MG cells. Conclusion： Netrin-4 activates the ERK / MAPK signaling pathway in glioblastoma cell lines,probably which protects GBM cell senescence clarifies the mechanism.