Phorbol 12-myristate 13-acetate (PMA)-induced migration of glioblastoma cells is mediated via p38MAPK/Hsp27 pathway
Phorbol 12-myristate 13-acetate (PMA)-induced migration of glioblastoma cells is mediated via p38MAPK/Hsp27 pathway
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DOI：
10.1016/j.bcp.2007.06.018
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发表时间：
2007-09-01
影响因子：
5.8
通讯作者：
Hamada, Jun-Ichiro
中科院分区：
文献类型：
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作者：
Nomura, Naoko;Nomura, Motohiro;Hamada, Jun-Ichiro
我们从p38丝裂原活化蛋白激酶(MAPK)/热休克蛋白27(Hsp27)通路入手，研究佛波酯(PMA)诱导胶质母细胞瘤细胞迁移的机制。PMA诱导A172胶质母细胞瘤细胞迁移及p38MAPK活化。用SB203580或小干扰RNA(SiRNA)抑制p38MAPK，可阻断PMA诱导的片状脂膜和斑状复合体的形成。此外，p38MAPK的激活导致F-肌动蛋白聚合调节因子Hsp27的磷酸化。免疫组织化学分析显示，在PMA刺激下，未磷酸化和磷酸化的HSP27均移位到片层脂膜。SB203580或p38MAPK siRNA阻断了这些现象，表明Hsp27的磷酸化和胞浆转位是由p38MAPK介导的。为了解决内源性Hsp27是否参与PMA诱导的迁移的问题，我们使用Hsp27 siRNA抑制Hsp27的表达。虽然siRNA敲除Hsp27对p38MAPK的激活影响不大，但片状脂膜和局灶性复合体的形成明显受到抑制。在Hsp27 siRNA转基因细胞中，迁移也被取消。总之，PMA诱导的迁移需要p38MAPK的激活和Hsp27的磷酸化。此外，Hsp27本身在PMA诱导的迁移中发挥了关键作用。我们的数据为一个模型提供了大量的证据，该模型阐明了PMA激活的蛋白激酶C在胶质母细胞瘤中调节肌动蛋白动态和迁移的分子机制。细胞。(C)2007 Elsevier Inc.保留所有权利。
We investigated the mechanism of phorbol 12-myristate 13-acetate (PMA)-induced migration of glioblastoma cells focusing on the p38 mitogen-activated protein kinase (MAPK)/heat shock protein 27 (Hsp27) pathway. PMA-induced cell migration and activation of p38MAPK in A172 glioblastoma cells. PMA-induced formation of lamellipodia and focal complexes was blocked by inhibiting p38MAPK with SB203580 or small interfering RNA (siRNA). Furthermore, activation of p38MAPK resulted in phosphorylation of an F-actin polymerization regulator, Hsp27. Immunohistochemical analysis showed that upon PMA stimulation, both unphosphorylated and phosphorylated Hsp27 were translocated to the lamellipodia. SB203580 or p38MAPK siRNA blocked these phenomena, indicating that Hsp27 phosphorylation and translocation from cytosol to membrane were mediated by p38MAPK. To address the question of whether endogenous Hsp27 participates in PMA-induced migration, we inhibited the expression of Hsp27 using Hsp27 siRNA. Although knockdown of Hsp27 by siRNA had little effect on p38MAPK activation, lamellipodia and focal complex formation was markedly inhibited. Migration was also abolished in Hsp27 siRNA-transfected cells. In conclusion, p38MAPK activation followed by Hsp27 phosphorylation was required for PMA-induced migration. Furthermore, Hsp27 itself played critical roles in PMA-induced migration. our data provide substantial evidence for a model elucidating the molecular mechanisms of regulation of actin dynamics and migration by PMA-activated protein kinase C in glioblastoma. cells. (C) 2007 Elsevier Inc. All rights reserved.