aFGF和Genistein对AGZY-83A细胞内PKC活性及[Ca^(2+)]i的影响

Effects of aFGF and Genistein on the PKC Activity and [Ca^(2+)]i in AGZY-83A Cells

目的 :探讨 a FGF及 TPK抑制剂 Genistein对 AGZY- 83A细胞内 PKC活性及游离 Ca2 +浓度的影响。方法 :用不同浓度的 a FGF和 Genistein诱导 AGZY- 83A细胞 ,(γ- P32 ) - ATP标记的 Histone 为底物 ,液体闪烁测定PKC活性 ;Fura- 2 / AM负载荧光光度法测定细胞内游离 Ca2 +浓度。结果 :a FGF诱导后 ,细胞内 PKC活性及 Ca2 +浓度升高 ,且与 a FGF呈剂量依赖效应。Genistein抑制细胞内 PKC活性及 Ca2 +浓度 ,也呈剂量依赖效应。Genistein对 a FGF诱导的 PKC活性抑制更显著。结论 :进一步证明 PKC和 Ca2 +确是 TPK的下游信号分子。

Objective: This paper was to investigate the effects of aFGF and TPK inhibitor Genistein on intracellular PKC activity and [Ca 2+ ]i in AGZY-83A cells. Methods: The activity of PKC in the cells were detected by the incorporation of (γ-P 32 )-ATP into exogenous substrate;[Ca 2+ ]i was measured with Fura-2/AM as fluorecence indicator. Results: The intracellular PKC activity increased with aFGF in a dose dependent manner. Genistein suppressed the intracellular PKC activity in a dose dependent manner. The highest percentage of the suppression on the cells which were treated by aFGF was 43%. After induced by aFGF, the [Ca 2+ ]i was increased in a dose-dependent manner. But Genistein decrease [Ca 2+ ]i in the cells, especially in the cells treated with aFGF. The highest percentage of inhibition on the cells treated with aFGF was 28%. Conclusion: aFGF might initiate a cascade of biochemical events, which ultimately cell proliferation by TPK receptor,PKC and Ca 2+ were located downstream of TPK in AGZY-83A Cells. Genistein can suppress the signal transduction pathway by inhibition of TPK.