PD98059 抑制aFGF诱导的人膀胱癌细胞株EJ细胞增殖

目的探讨细胞外信号调节激酶(ERK)激酶MEK抑制剂PD98059对酸性成纤维细胞生长因子(aFGF)诱导的人膀胱癌细胞株EJ细胞增殖的抑制作用。方法以不同浓度的aFGF和PD98059作用EJ细胞,通过细胞计数法、MTT比色法观察PD98059对细胞增殖的抑制作用;流式细胞仪测定细胞各周期细胞百分数的变化;利用γ3-2P]ATP掺入外源性底物的方法,液体闪烁测定ERK活性的变化。结果aFGF使EJ细胞株增殖比明显增加,而PD98059使EJ细胞株增殖比明显下降,在一定浓度范围内,其程度均随浓度增高而增强。EJ细胞受到aFGF刺激后,由G0+G1期进入S和G2+M期的细胞明显增多,在PD98059的作用下,由G0+G1期进入S和G2+M期的细胞明显减少,并在一定浓度范围内成剂量依赖性,与对照组比较差异显著(P<0.05);随着aFGF浓度的增加,EJ细胞ERK活性增高,PD98059抑制细胞内ERK活性,与PD98059浓度呈剂量依赖效应。结论PD98059对aFGF引起的EJ细胞增殖具有抑制作用,aFGF可能通过激活Ras-Raf-ERK信号转导途径来调控EJ细胞增殖,PD98059可有效阻滞此传导途径。

Inhibitory Effect of PD98059 on the Proliferation of EJ Cells Induced by aFGF

Objective To explore the inhibitory effect of extracellular regulated protein kinase(ERK) inhibitor PD98059 on the proliferation of EJ cells induced by acidic fibroblast growth factor(aFGF).Methods Cytometry and MTT assay were used to determine the suppressive effect of PD98059 on EJ cells induced by different concentration of aFGF.The percentage of different cell cycles was measured by Flow cytometry.The activity of ERK in cells induced by different concentration of aFGF and PD98059 was detected by incorporation of _ATP into exogenous substrate.Results aFGF promoted the proliferation of EJ cells significantly,and PD98059 suppressed the proliferation of EJ cells significantly.The percentage of cell cycles from phase G_(0) G_(1) to S and G_(2) M increased by aFGF significantly.The percentage of cell cycles from phase G_(0) G_(1) to S and G_(2) M decreased by PD98059 significantly.The intracellular ERK activity increased by aFGF,PD98059 suppressed the intracellular ERK activity,especially in the cells treated with aFGF.Conclusion PD98059 suppressed the proliferation of EJ cells induced by aFGF by blocking the activation of Ras-Raf-ERK signal transductive pathway.