FRK通过抑制ERK信号通路对脑胶质瘤细胞增殖作用的研究

摘要

Objective To study the mechanism of FRK regulating glioma cells proliferation through inhibiting ERK signaling pathway .Methods FRK plasmid was transfected into U 251 cells by PolyJetTM.Western blot(WB) was applied to detect the efficiency of FRK over-expression and P-ERK, ERK protein levels, and EDU incorporation assay was used to explore the effect of FRK over-expression on glioma U251 cells proliferation.After treatment with ERK inhibitor PD98059, the protein level of FRK , P-ERK and ERK was tested by WB , and the proliferation ability of glioma U251 cells was examined by EDU incorporation assay .Results WB results showed that the FRK plasmid was transfected into glioma U 251 cells successfully , FRK over-expression decreased the proliferation ability of glioma U251 cells.And FRK over-expression decreased the protein level of P-ERK, but had no effect on the protein level of ERK . Compared with control group , the phosphorylation of ERK decreased significantly in PD 98059 treatment group , but the protein level of FRK had no change.The proliferation ability of U251 cells was significantly decreased after treatment with ERK inhibitor PD98059.Conclusion FRK may regulate the proliferation of glioma U251 cells via inhibiting ERK pathway .%目的 研究FRK是否通过抑制ERK信号通路进而影响脑胶质瘤细胞的增殖.方法 应用PolyJetTM将FRK质粒转染入脑胶质瘤U251细胞中,Western blot(WB)检测转染效率及P-ERK、ERK蛋白水平的变化,EDU实验观察脑胶质瘤细胞增殖能力的变化;用ERK抑制剂PD98059处理U251细胞,WB检测细胞中FRK、P-ERK、ERK的蛋白水平,EDU实验检测脑胶质瘤细胞增殖能力的变化.结果 WB检测显示FRK质粒转染成功,过表达FRK使U251细胞增殖能力降低.过表达FRK降低了P-ERK的蛋白水平,但对ERK总蛋白水平无影响.与对照组相比,ERK抑制剂PD98059组P-ERK的蛋白水平明显降低,但对FRK的蛋白水平无明显影响.ERK抑制剂PD98059处理后,脑胶质瘤U251细胞增殖能力明显降低.结论 FRK可以通过抑制ERK的活性,从而降低脑胶质瘤细胞的增殖.