Serum-Activated K and Cl Currents Underlay U87-MG Glioblastoma Cell Migration
Serum-Activated K and Cl Currents Underlay U87-MG Glioblastoma Cell Migration
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DOI：
10.1002/jcp.22523
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发表时间：
2011-07-01
影响因子：
5.6
通讯作者：
Franciolini, Fabio
中科院分区：
文献类型：
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作者：
Catacuzzeno, Luigi;Aiello, Francesco;Franciolini, Fabio
胶质母细胞瘤细胞在体内暴露于多种促迁移信号，包括由于血脑屏障破坏而渗入高级别胶质瘤的未定义血清成分。胶质母细胞瘤细胞迁移已进一步证明严重依赖于离子通道的活性。然后，我们研究了胎牛血清（FCS）对离子通道的调节作用，以及它们在U87-MG细胞迁移中的作用。使用穿孔膜片钳技术，我们发现，在一个细胞亚群（42%）中，FCS诱导：(1)TRAM-34敏感的、中等电导的钙活化K （IKCa）通道的振荡活性，由钙振荡介导，之前在该细胞系中被证明是由FCS诱导的；(2)对DIDS和nppb敏感的Cl电流稳定激活，显示出向外整流的瞬时电流-电压关系和缓慢的电压依赖性失活。相比之下，在另一个细胞亚群（32%）中，FCS诱导了单一的、短暂的IKCa电流激活，总是伴随着Cl电流的稳定激活。其余细胞对FCS无反应。为了了解fcs诱导的离子通道活性是否有助于促进细胞迁移，我们使用transwell迁移实验测试了TRAM-34和DIDS对fcs诱导的U87-MG细胞迁移的影响。我们发现这些抑制剂能够在FCS存在的情况下显著减少U87-MG细胞的迁移，并且它们的共同应用几乎完全阻止了迁移。结果表明，在fcs诱导的胶质母细胞瘤细胞迁移过程中，钾离子和氯离子通量的调节是必不可少的。j .细胞。物理学报，26(6):926- 933,2011。(C) 2010 Wiley-Liss, Inc。
Glioblastoma cells in vivo are exposed to a variety of promigratory signals, including undefined serum components that infiltrate into high grade gliomas as result of blood-brain barrier breakdown. Glioblastoma cell migration has been further shown to depend heavily on ion channels activity. We have then investigated the modulatory effects of fetal calf serum (FCS) on ion channels, and their involvement in U87-MG cells migration. Using the perforated patch-clamp technique we have found that, in a subpopulation of cells (42%), FCS induced: (1) an oscillatory activity of TRAM-34 sensitive, intermediate-conductance calcium-activated K (IKCa) channels, mediated by calcium oscillations previously shown to be induced by FCS in this cell line; (2) a stable activation of a DIDS- and NPPB-sensitive Cl current displaying an outward rectifying instantaneous current-voltage relationship and a slow, voltage-dependent inactivation. By contrast, in another subpopulation of cells (32%) FCS induced a single, transient IKCa current activation, always accompanied by a stable activation of the Cl current. The remaining cells did not respond to FCS. In order to understand whether the FCS-induced ion channel activities are instrumental to promoting cell migration, we tested the effects of TRAM-34 and DIDS on the FCS-induced U87-MG cell migration using transwell migration assays. We found that these inhibitors were able to markedly reduce U87-MG cell migration in the presence of FCS, and that their co-application resulted in an almost complete arrest of migration. It is concluded that the modulation of K and Cl ion fluxes is essential for the FCS-induced glioblastoma cell migration. J. Cell. Physiol. 226: 1926-1933, 2011. (C) 2010 Wiley-Liss, Inc.