Neuropilin-1-mediated vascular permeability factor/vascular endothelial growth factor-dependent endothelial cell migration
Neuropilin-1-mediated vascular permeability factor/vascular endothelial growth factor-dependent endothelial cell migration
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DOI：
10.1074/jbc.m310047200
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发表时间：
2003-12-05
影响因子：
4.8
通讯作者：
Mukhopadhyay, D
中科院分区：
文献类型：
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作者：
Wang, L;Zeng, HY;Mukhopadhyay, D
Neuropilin-1 (NRP-1) 已被发现在内皮细胞和肿瘤细胞中表达，作为血管通透因子/血管内皮生长因子 (VEGF) 的亚型特异性受体。之前的研究主要集中在NRP-1的胞外结构域上，该结构域可以与VEGF（165）结合，从而使NRP-1能够充当VEGF（165）的共受体，从而增强其与VEGFR-2的结合及其生物活性。然而，NRP-1 在血管生成中的确切功能作用和相关信号机制尚不清楚。在本研究中，我们通过将表皮生长因子受体的胞外结构域与NRP-1的跨膜和胞内结构域融合，构建了嵌合受体EGNP-1，并用逆转录病毒表达载体将其转导到HUVEC中。我们观察到 NRP-1/EGNP-1 介导配体刺激的人脐静脉内皮细胞 (HUVEC) 迁移，但不介导增殖。我们的结果表明，NRP-1 单独可以通过其胞内结构域介导 HUVEC 迁移，并且其 C 端三个氨基酸 (SEA-COOH) 对于该过程至关重要。我们证明磷脂酰肌醇 3 激酶抑制剂 Ly294002 和 p85 显性失活突变体可以阻断 NRP-1 介导的 HUVEC 迁移。 RhoA 显性失活突变体 (RhoA-19N) 的过表达可显着减少 NRP-1 介导的迁移。此外，NRP-1 介导的 HUVEC 迁移也需要 G(q) 家族蛋白和 Gbetagamma 亚基。这些结果首次表明NRP-1可以独立促进内皮细胞中的细胞信号传导，并且还证明了NRP-1的最后三个氨基酸对其功能的重要性。
Neuropilin-1 (NRP-1) has been found to be expressed by endothelial cells and tumor cells as an isoform-specific receptor for vascular permeability factor/vascular endothelial growth factor (VEGF). Previous studies were mainly focused on the extracellular domain of NRP-1 that can bind to VEGF(165) and, thus, enables NRP-1 to act as a co-receptor for VEGF(165), which enhances its binding to VEGFR-2 and its bioactivity. However, the exact functional roles and related signaling mechanisms of NRP-1 in angiogenesis are not well understood. In this study we constructed a chimeric receptor, EGNP-1, by fusing the extracellular domain of epidermal growth factor receptor to the transmembrane and intracellular domains of NRP-1 and transduced it into HUVECs with a retroviral expression vector. We observed that NRP-1/EGNP-1 mediates ligand-stimulated migration of human umbilical vein endothelial cells (HUVECs) but not proliferation. Our results show that NRP-1 alone can mediate HUVEC migration through its intracellular domain, and its C-terminal three amino acids (SEA-COOH) are essential for the process. We demonstrate that phosphatidylinositol 3-kinase inhibitor Ly294002 and the p85 dominant negative mutant can block NRP-1-mediated HUVEC migration. NRP-1-mediated migration can be significantly reduced by overexpression of the dominant negative mutant of RhoA (RhoA-19N). In addition, G(q) family proteins and Gbetagamma subunits are also required for NRP-1-mediated HUVEC migration. These results show for the first time that NRP-1 can independently promote cell signaling in endothelial cells and also demonstrate the importance of last three amino acids of NRP-1 for its function.