Sphingosine-1-phosphate induces VEGF-C expression through a MMP-2/FGF-1/FGFR1-dependent pathway in endothelial cells in vitro
Sphingosine-1-phosphate induces VEGF-C expression through a MMP-2/FGF-1/FGFR1-dependent pathway in endothelial cells in vitro
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DOI：
10.1038/aps.2012.186
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发表时间：
2013-03-01
影响因子：
8.2
通讯作者：
Lee, Hsinyu
中科院分区：
文献类型：
--
作者：
Chang, Chi-hao;Huang, Yuan-li;Lee, Hsinyu
目的：目的：探讨1-磷酸鞘氨醇（sphingosine-1-phosphate，SIP）能否诱导血管内皮细胞表达血管内皮生长因子-C（vascular endothelial growth factor-C，VEGF-C），并探讨其作用机制。使用实时PCR评估细胞中VEGF-C mRNA的表达。用ELISA法检测细胞中VEGF-C蛋白和FGFR-I磷酸化。结果：1、5、10 μ mol/L的SIP能显著上调HUVECs中VEGF-C的表达，并能显著下调MMP-2、FGF-1和FGFR-1的表达。MMP抑制剂GM 6001或FGFR抑制剂SU 5402预处理可阻断该作用，但EGFR抑制剂AGI 478不能阻断该作用。在用FGFR-1或MMP-2 siRNA转染的HUVEC中，该作用也被阻断。此外，与SIP（5 μ mol/L）孵育的HUVECs显着增加FGFR-I磷酸化，这是由GM 6001阻断。结论：SIP通过MMP-2/ FGF-1/FGFR-1依赖性途径诱导VEGF-C表达。
Aim: To investigate whether sphingosine-1-phosphate (SIP), a potent angiogenic factor, induced vascular endothelial growth factor-C (VEGF-C) expression in endothelial cells in vitro and to examine its underlying mechanisms.Methods: Human umbilical vein endothelial cells (HUVECs) were examined. VEGF-C mRNA expression in the cells was assessed using real-time PCR. VEGF-C protein and FGFR-I phosphorylation in the cells were measured with ELISA. RNA interference was used to downregulate the expression of matrix metalloproteinase-2 (MMP-2), fibroblast growth factor-1 (FGF-1) and FGF receptor-1 (FGFR-1).Results: Incubation of HUVECs with SIP (1, 5, and 10 mu mol/L) significantly increased VEGF-C expression. The effect was blocked by pretreatment with the MMP inhibitor GM6001 or the FGFR inhibitor SU5402, but not the EGFR inhibitor AGI478. The effect was also blocked in HUVECs that were transfected with FGFR-1 or MMP-2 siRNA. Furthermore, incubation of HUVECs with SIP (5 mu mol/L) significantly increased FGFR-I phosphorylation, which was blocked by GM6001. Moreover, knockdown of FGF-1, not FGF-2, in HUVECs with siRNAs, blocked SIP-induced VEGF-C expression.Conclusion: SIP induces VEGF-C expression through a MMP-2/ FGF-1/FGFR-1-dependent pathway in HUVECs.