FMLP-induced formation of F-actin in HL60 cells is dependent on PI3-K but not on intracellular Ca2+, PKC, ERK or p38 MAPK
FMLP-induced formation of F-actin in HL60 cells is dependent on PI3-K but not on intracellular Ca2+, PKC, ERK or p38 MAPK
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DOI：
10.1007/s000110050647
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发表时间：
2000-12-01
影响因子：
6.7
通讯作者：
Kuwabara, M
中科院分区：
文献类型：
--
作者：
Cui, YD;Inanami, O;Kuwabara, M
目标与设计：进一步了解F-actin形成的信号转导途径的机制本文研究了吞噬细胞内肌动蛋白（actin）的聚合和NADPH氧化酶的激活，以及各种抑制剂对它们的影响。研究分化的HL 60细胞以检查它们的N-甲酰基-甲硫氨酰基-亮氨酰基-苯丙氨酸（fMLP）刺激的F-肌动蛋白和NADPH氧化酶的激活后，用各种抑制剂处理。其中包括蛋白激酶C（PKC）抑制剂（GF 109203 X），磷脂酰肌醇3激酶（PI 3-K）抑制剂（渥曼青霉素），一种细胞外反应激酶（ERK）抑制剂（PD 98059），一种p38丝裂原活化蛋白激酶（MAPK）抑制剂结果：Wortmannin处理后，F-actin的形成受到抑制，而NADPH氧化酶的激活受到的抑制较小。BAPTA-AM和GF 109203 X不减弱F-肌动蛋白的形成，但完全抑制NADPH氧化酶的活化。PD 98059和SE 203580部分抑制NADPH氧化酶的活化，但不影响F-肌动蛋白的形成。此外，wortmannin而不是BAPTA-AM和GF 109203 X抑制了fMLP诱导的Akt活化，已知Akt调节NADPH氧化酶。这些结果表明，F-actin的形成依赖于PI 3-K，而不依赖于PKC、ERK和p38 MAPK，也不依赖于细胞内Ca ~（2+）的增加，而NADPH氧化酶的激活则部分依赖于ERK、p38 MAPK，Akt受PI 3-K的调节，并强烈依赖于PKC的激活和细胞内Ca ~（2+）的增加。
Objective and Design: To further understand the mechanisms of signal transduction pathways for the formation of F-actin (polymerization of actin) and the activation of NADPH oxidase in phagocytic cells, the effects of various inhibitors on them were studied.Materials and Methods: Differentiated HL60 cells were studied to examine their N-formyl-methionyl-leucyl-phenyl-alanine (fMLP)-stimulated formation of F-actin and activation of NADPH oxidase following treatment with various inhibitors. These included a protein kinase C (PKC) inhibitor (GF 109203X), a phosphatidylinositide 3 kinase (PI3-K) inhibitor (wortmannin), an extracellular response kinase (ERK) inhibitor (PD 98059), a p38 mitogen-activated protein kinase (MAPK) inhibitor (SB 203580) and an intracellular Ca2+-chelator (BAPTA-AM).Results: The treatment with wortmannin suppressed the formation of F-actin, with less suppression of the activation of NADPH oxidase. BAPTA-AM and GF 109203X did not attenuate the formation of F-actin but completely inhibited the activation of NADPH oxidase. PD 98059 and SE 203580 partially inhibited the activation of NADPH oxidase without influence on the formation of F-actin. Furthermore, wortmannin but not BAPTA-AM and GF 109203X inhibited the fMLP-induced activation of Akt, which is known to regulate NADPH oxidase.Conclusions: These results suggest that the formation of F-actin is dependent on PI3-K and independent of PKC, ERK and p38 MAPK as well as the increase in intracellular Ca2+, whereas the activation of NADPH oxidase is partly dependent on ERK, p38 MAPK, Akt regulated by PI3-K, and strongly dependent on the activation of PKC and the increase in intracellular Ca2+.