MRP-1/CD9 gene transduction regulates the actin cytoskeleton through the downregulation of WAVE2
MRP-1/CD9 gene transduction regulates the actin cytoskeleton through the downregulation of WAVE2
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DOI：
10.1038/sj.onc.1209654
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发表时间：
2006-10-19
期刊：
影响因子：
8
通讯作者：
Miyake, M.
中科院分区：
文献类型：
--
作者：
Huang, C-L;Ueno, M.;Miyake, M.
运动相关蛋白-1（MRP-1/CD 9）参与细胞运动。我们研究了MRP-1/CD 9基因转染HT 1080细胞后肌动蛋白细胞骨架的变化，以及肌动蛋白相关蛋白（阿普）2和Arp 3以及Wiskott-Aldrich综合征蛋白（WASP）家族的表达。MRP-1/CD 9转染的HT 1080细胞中板状伪足细胞的出现率明显低于对照组（P < 0.0001）。MRP-1/CD 9基因转导影响Arp 2和Arp 3蛋白的亚细胞定位。MRP-1/CD 9基因转导后WAVE 2表达下调（P < 0.0001）。而Arp 2、Arp 3及其他WASP的表达无明显差异。抗MRP-1/CD 9单克隆抗体可抑制MRP-1/CD 9转染的HT 1080细胞WAVE 2表达下调（P < 0.0001），逆转MRP-1/CD 9基因转导的形态学效应。此外，通过转染WAVE 2特异性小干扰RNA（siRNA）下调WAVE 2模拟MRP-1/CD 9基因转导的形态学效应并抑制细胞运动。然而，转染Wnt 1、Wnt 2b 1或Wnt 5a的每种siRNA均不影响WAVE 2的表达。转染WAVE 2特异性siRNA也不影响这些Wnt的表达。这些结果表明，MRP-1/CD 9通过Wnt非依赖性信号通路下调WAVE 2来调节肌动蛋白细胞骨架。
Motility-related protein-1 (MRP-1/CD9) is involved in cell motility. We studied the change in the actin cytoskeleton, and the expression of actin-related protein (Arp) 2 and Arp3 and the Wiskott-Aldrich syndrome protein ( WASP) family according to MRP-1/CD9 gene transduction into HT1080 cells. The frequency of cells with lamellipodia was significantly lower in MRP-1/CD9-transfected HT1080 cells than in control HT1080 cells (P < 0.0001). MRP-1/CD9 gene transduction affected the subcellular localization of Arp2 and Arp3 proteins. Furthermore, MRP-1/CD9 gene transduction induced a downregulation of WAVE2 expression (P < 0.0001). However, no difference was observed in the expression of Arp2, Arp3 or other WASPs. A neutralizing anti-MRP-1/CD9 monoclonal antibody inhibited downregulation of WAVE2 in MRP-1/CD9-transfected HT1080 cells (P < 0.0001 ), and reversed the morphological effects of MRP-1/CD9 gene transduction. Furthermore, downregulation of WAVE2 by transfection of WAVE2-specific small interfering RNA ( siRNA) mimicked the morphological effects of MRP-1/CD9 gene transduction and suppressed cell motility. However, transfection of each siRNA for Wnt1, Wnt2b1 or Wnt5a did not affect WAVE2 expression. Transfection of WAVE2-specific siRNA also did not affect expressions of these Wnts. These results indicate that MRP-1/CD9 regulates the actin cytoskeleton by downregulating of the WAVE2, through the Wnt-independent signal pathway.