aFGF对人卵巢癌细胞株(CAOV_3)TPK、PKC及ERK活性的影响

Effects of aFGF on the Activity of TPK, PKC and ERK in CAOV_3 Cell Line

目的探讨酸性成纤维细胞生长因子(aFGF)和细胞外信号调节激酶(ERK)丝裂原活化蛋白激酶激酶(MEK)抑制剂PD98059对人卵巢上皮性癌细胞株(CAOV_3)细胞内酪氨酸蛋白激酶(TPK)、蛋白激酶C(PKC)及ERK活性的影响。方法不同浓度aFGF和PD98059诱导CAOV_3细胞,[γ-^(32)P]ATP掺入外源性底物,液体闪烁测定TPK、PKC及ERK活性的变化;Western印迹方法检测ERK表达水平。结果随着aFGF浓度增加,CAOV_3细胞内TPK、PKC及ERK活性随之升高,与aFGF浓度呈剂量依赖效应;PD98059抑制细胞内PKC及ERK活性,与PD98059浓度亦呈剂量依赖效应,并且PD98059对aFGF诱导的PKC及ERK活性抑制更显著(P<0.05)。Western印迹方法检测ERK表达水平与上述结果基本一致。结论aFGF可能通过TPK途径激活PKC及ERK来调控CAOV_3细胞增殖,PKC及ERK是TPK下游信号分子,二者处于同一信号通路上不同环节。

Objective To explore the effects of the aFGF and PD98059 on the activity of TPK, PKC and ERK in CAOV_3 cell line. Methods The activity of TPK, PKC and ERK in cells induced by different concentration of aFGF and PD98059 was detected by incorporation of [γ-^(32)P]-ATP into exogenous substrate. The expression of ERK induced by aFGF and PD98059 was detected by Western blot. Results The intracellular TPK, PKC and ERK activity increased with aFGF, and PD98059 suppressed the activities of intracellular PKC and ERK, especially in the cells treated with aFGF (P<0.05). PD98059 had no effect on the activity of TPK. The Western blot yielded similar results. Conclusion aFGF might adjust the proliferation of CAOV_3 by activating PKC and ERK through TPK signal transduction pathway, and PKC and ERK are located in the downstream part of TPK in CAOV3 cell line.