bFGF对NIH3T3细胞TPK,PKC,ERK活性的影响

Effect of bFGF on activities of TPK, PKC, and ERK of NIH3T3 cell line

目的 :观察碱性成纤维细胞生长因子 (bFGF)对NIH3T3细胞酪氨酸蛋白激酶 (TPK)、蛋白激酶C(PKC)及胞外调节蛋白激酶 (ERK)活性的影响 ,探讨其信号转导机制。方法 :分别加入bFGF及PKC抑制剂H 7、MEK1抑制剂PD980 5 9孵育细胞 ,利用 [γ 3 2 P]ATP掺入外源性底物的方法测定活性。结果 :bFGF可使细胞膜TPK、细胞质PKC及ERK活性明显升高。H 7+bFGF、PD980 5 9+bFGF组与只加bFGF组比较TPK活性保持不变 ,而PKC及ERK活性均下降。结论 :TPK、PKC、ERK介导bFGF的信号转导。ERK与PKC是TPK受体下游的两个分支 。

Objective: To examine the effect of basic fibroblast growth factor (bFGF) of different concentrations on activitites of tyrosine protein kinase (TPK), protein kinase C(PKC) and extracellular signal regulated protein kinase (ERK) in NIH3T3 cell line and to elucidate the signal pathway in cell proliferation and differentiation. Methods:The bFGF, H 7 and PD98059 were added separately to incubate cell. Isotope was used to check activity. Results: The activities of membrane TPK, cytosolic PKC, and ERK increased obviously after bFGF treatment compared with the control group. In H 7 and PD98059 groups, the activity of TPK was the same as bFGF group, while the activities of PKC and ERK decreased. Conclusion:The TPK, PKC, and ERK mediate bFGF signal transduction in NIH3T3 cell line. The PKC and ERK are two different downstream pathways of TPK receptor. Two pathways activate each other.