Sphingosine 1-phosphate stimulates cell migration through a Gi-coupled cell surface receptor -: Potential involvement in angiogenesis
Sphingosine 1-phosphate stimulates cell migration through a Gi-coupled cell surface receptor -: Potential involvement in angiogenesis
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DOI：
10.1074/jbc.274.50.35343
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发表时间：
1999-12-10
影响因子：
4.8
通讯作者：
Spiegel, S
中科院分区：
文献类型：
--
作者：
Wang, F;Van Brocklyn, JR;Spiegel, S
Sphingosine l-phosphate （SPP）已被证明抑制多种细胞的趋化性，在某些情况下是通过细胞内作用，而在其他情况下是通过受体介导的作用。令人惊讶的是，我们发现低浓度的SPP （10-100 nm）增加了过表达G蛋白偶联SPP受体EDG-1的HEK293细胞的趋化性。与先前在人类乳腺癌细胞中的发现一致（Wang， F., Nohara， K., Olivera， O., Thompson， E. W., and Spiegel， S. (1999) Exp. Cell Res. 247, 17-28）， SPP在微摩尔浓度下，抑制了载体和edg -1过表达的HEK293细胞的趋化性。纳摩尔浓度的SPP对表达SPP受体EDG-1和EDG-3的人脐静脉内皮细胞（HUVEC）和牛主动脉内皮细胞（BAEC）的趋化性也有显著的促进作用，而较高浓度的SPP作用较弱。百日毒可以使adp核糖化并使G(i)偶联受体失活，从而阻断spp诱导的趋化作用。棋盘分析表明SPP刺激趋化和趋化作用。综上所述，这些数据表明SPP通过结合EDG-1刺激细胞迁移。与SPP类似，sph-inganine 1-phosphate（二氢-SPP）也与SPP受体家族结合，增强趋化性；然而，另一种结构相关的溶血磷脂，溶血磷脂酸，不与SPP竞争结合，也对内皮细胞的趋化性没有显著影响。此外，SPP以百日咳毒素敏感的方式增加了HUVEC和BAEC的增殖。SPP和二氢-SPP还能促进BAEC在胶原凝胶上的成管（体外血管生成），并增强碱性成纤维细胞生长因子诱导的成管。百日咳毒素处理可阻断SPP-，但不能阻断bfgf刺激的体外血管生成。我们的研究结果表明SPP可能通过与内皮细胞gi偶联SPP受体结合而在血管生成中发挥作用。
Sphingosine l-phosphate (SPP) has been shown to inhibit chemotaxis of a variety of cells, in some cases through intracellular actions, while in others through receptor-mediated effects. Surprisingly, we found that low concentrations of SPP (10-100 nm) increased chemotaxis of HEK293 cells overexpressing the G protein-coupled SPP receptor EDG-1. In agreement with previous findings in human breast cancer cells (Wang, F., Nohara, K., Olivera, O., Thompson, E. W., and Spiegel, S. (1999) Exp. Cell Res. 247, 17-28), SPP, at micromolar concentrations, inhibited chemotaxis of both vector- and EDG-1-overexpressing HEK293 cells. Nanomolar concentrations of SPP also induced a marked increase in chemotaxis of human umbilical vein endothelial cells (HUVEC) and bovine aortic endothelial cells (BAEC), which express the SPP receptors EDG-1 and EDG-3, while higher concentrations of SPP were less effective. Treatment with pertussis toxin, which ADP-ribosylates and inactivates G(i)-coupled receptors, blocked SPP-induced chemotaxis. Checkerboard analysis indicated that SPP stimulates both chemotaxis and chemokinesis. Taken together, these data suggest that SPP stimulates cell migration by binding to EDG-1. Similar to SPP, sph-inganine 1-phosphate (dihydro-SPP), which also binds to this family of SPP receptors, enhanced chemotaxis; whereas, another structurally related lysophospholipid, lysophosphatidic acid, did not compete with SPP for binding nor did it have significant effects on chemotaxis of endothelial cells. Furthermore, SPP increased proliferation of HUVEC and BAEC in a pertussis toxin-sensitive manner. SPP and dihydro-SPP also stimulated tube formation of BAEC grown on collagen gels (in vitro angiogenesis), and potentiated tube formation induced by basic fibroblast growth factor. Pertussis toxin treatment blocked SPP-, but not bFGF-stimulated in vitro angiogenesis. Our results suggest that SPP may play a role in angiogenesis through binding to endothelial cell Gi-coupled SPP receptors.