Involvement of RhoA/Rho kinase signaling in VEGF-induced endothelial cell migration and angiogenesis in vitro
Involvement of RhoA/Rho kinase signaling in VEGF-induced endothelial cell migration and angiogenesis in vitro
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DOI：
10.1161/01.atv.0000054198.68894.88
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发表时间：
2003-02-01
影响因子：
8.7
通讯作者：
van Hinsbergh, VWM
中科院分区：
文献类型：
--
作者：
Amerongen, GPV;Koolwijk, P;van Hinsbergh, VWM
目的生长因子诱导的血管生成涉及内皮细胞 (EC) 迁移到血管周围区域，并且需要主动重塑内皮 F-肌动蛋白细胞骨架。小GTP酶RhoA先前被认为参与血管内皮生长因子(VEGF)诱导的信号通路，但其作用尚未阐明。方法和结果-VEGF诱导RhoA的激活并将RhoA募集到人EC的细胞膜上。 RhoA 活性的增加对于 VEGF 诱导的 F-肌动蛋白细胞骨架重组是必要的，如显性失活 RhoA 的腺病毒转染所证明的。 Rho 激酶介导 RhoA 的这种作用，正如使用 Rho 激酶特异性抑制剂 Y-27632 所证明的那样。抑制 Rho 激酶可阻止 VEGF 增强的 EC 迁移以响应机械损伤，但对基础 EC 迁移没有影响。此外，在体外血管生成模型中，抑制 RhoA 或 Rho 激酶可减弱 VEGF 介导的 EC 在 3 维纤维蛋白基质中的向内生长。 结论：VEGF 诱导的 EC 细胞骨架变化需要 RhoA 和 Rho 激酶，并且 RhoA/Rho 激酶信号传导的激活参与 VEGF 诱导的体外 EC 迁移和血管生成。
Objective-Growth factor-induced angiogenesis involves migration of endothelial cells (ECs) into perivascular areas and requires active remodeling of the endothelial F-actin cytoskeleton. The small GTPase RhoA previously has been implicated in vascular endothelial growth factor (VEGF)-induced signaling pathways, but its role has not been clarified.Methods and Results-VEGF induced the activation of RhoA and recruited RhoA to the cell membrane of human ECs. This increase in RhoA activity is necessary for the VEGF-induced reorganization of the F-actin cytoskeleton, as demonstrated by adenoviral transfection of dominant-negative RhoA. Rho kinase mediated this effect of RhoA, as was demonstrated by the use of Y-27632, a specific inhibitor of Rho kinase. Inhibition of Rho kinase prevented the VEGF-enhanced EC migration in response to mechanical wounding but had no effect on basal EC migration. Furthermore, in an in vitro model for angiogenesis, inhibition of either RhoA or Rho kinase attenuated the VEGF-mediated ingrowth of ECs in a 3-dimensional fibrin matrix.Conclusions-VEGF-induced cytoskeletal changes in ECs require RhoA and Rho kinase, and activation of RhoA/Rho kinase signaling is involved in the VEGF-induced in vitro EC migration and angiogenesis.