Neuropilin-1 mediates PDGF stimulation of vascular smooth muscle cell migration and signalling via p130Cas.
Neuropilin-1 mediates PDGF stimulation of vascular smooth muscle cell migration and signalling via p130Cas.
复制标题
DOI：
10.1042/bj20100580
复制
发表时间：
2011-05-01
期刊：
影响因子：
--
通讯作者：
Zachary IC
中科院分区：
文献类型：
--
作者：
Pellet-Many C;Frankel P;Evans IM;Herzog B;Jünemann-Ramírez M;Zachary IC
NRP1（neuropilin-1）是内皮细胞中 VEGF（血管内皮生长因子）家族成员的共同受体，但越来越多地参与其他生长因子诱导的信号传导。 NRP1 在 VSMC（血管平滑肌细胞）中表达，但其功能和相关机制尚不清楚。本研究旨在确定 NRP1 在 HCASMC（人冠状动脉平滑肌细胞）对 PDGF（血小板源性生长因子）的迁移反应中的作用，并确定所涉及的信号传导机制。 NRP1 在 HAoSMC（人主动脉平滑肌细胞）和 HCASMC 中高表达，并在 VSMC 中通过富含 CS（硫酸软骨素）的 Ser612 位点 O 联糖基化进行修饰。由 PDGF-BB 和 PDGF-AA 诱导的 HCASMC 迁移受到 NRP1 siRNA（小干扰 RNA）和缺乏胞内结构域的 NRP1 突变体 (Ad.NRP1ΔC) 的腺病毒过表达的抑制。 NRP1 与 PDGFRα（PDGF 受体 α）进行免疫共沉淀，免疫荧光染色表明 NRP1 和 PDGFRα 共定位于 VSMC。 NRP1 siRNA 还抑制 PDGF 诱导的 PDGFRα 激活。 NRP1 特异性 siRNA、Ad.NRP1ΔC 和使用软骨素酶去除 CS 聚糖均抑制 PDGF-BB 和 -AA 对接头蛋白 p130Cas（Cas 是 Crk 相关底物）酪氨酸磷酸化的刺激，对其他主要信号传导途径几乎没有影响，并且 p130Cas 敲低抑制 HCASMC 迁移。 PDGF 诱导的趋化性和 p130Cas 磷酸化被软骨素酶抑制，此外，不可糖基化的 NRP1S612A 突变体的腺病毒表达抑制趋化性，但不抑制 p130Cas 磷酸化。这些结果表明 NRP1 和 NRP1 糖基化在介导 PDGF 诱导的 VSMC 迁移中的作用，可能是通过充当 PDGFRα 的共受体并通过选择性动员新型 p130Cas 酪氨酸磷酸化途径。
NRP1 (neuropilin-1) is a co-receptor for members of the VEGF (vascular endothelial growth factor) family in endothelial cells, but is increasingly implicated in signalling induced by other growth factors. NRP1 is expressed in VSMCs (vascular smooth muscle cells), but its function and the mechanisms involved are poorly understood. The present study aimed to determine the role of NRP1 in the migratory response of HCASMCs (human coronary artery smooth muscle cells) to PDGF (platelet-derived growth factor), and to identify the signalling mechanisms involved. NRP1 is highly expressed in HAoSMCs (human aortic smooth muscle cells) and HCASMCs, and modified in VSMCs by CS (chondroitin sulfate)-rich O-linked glycosylation at Ser612. HCASMC migration induced by PDGF-BB and PDGF-AA was inhibited by NRP1 siRNA (small interfering RNA), and by adenoviral overexpression of an NRP1 mutant lacking the intracellular domain (Ad.NRP1ΔC). NRP1 co-immunoprecipitated with PDGFRα (PDGF receptor α), and immunofluorescent staining indicated that NRP1 and PDGFRα co-localized in VSMCs. NRP1 siRNA also inhibited PDGF-induced PDGFRα activation. NRP1-specific siRNA, Ad.NRP1ΔC and removal of CS glycans using chondroitinase all inhibited PDGF-BB and -AA stimulation of tyrosine phosphorylation of the adapter protein, p130Cas (Cas is Crk-associated substrate), with little effect on other major signalling pathways, and p130Cas knockdown inhibited HCASMC migration. Chemotaxis and p130Cas phosphorylation induced by PDGF were inhibited by chondroitinase, and, additionally, adenoviral expression of a non-glycosylatable NRP1S612A mutant inhibited chemotaxis, but not p130Cas phosphorylation. These results indicate a role for NRP1 and NRP1 glycosylation in mediating PDGF-induced VSMC migration, possibly by acting as a co-receptor for PDGFRα and via selective mobilization of a novel p130Cas tyrosine phosphorylation pathway.