肝细胞生长因子通过鞘氨醇激酶途径诱导内皮细胞迁移作用的研究

目的：阐明鞘氨醇激酶（SPK）在肝细胞生长因子（HGF）诱导的内皮细胞迁移中的调节作用。方法：构建携带野生型SPK（SPK^WT）及负显性SPK（SPKDN）基因的重组腺病毒载体并包装获得重组腺病毒;用重组腺病毒感染ECV304细胞,检测感染效率及目的基因的表达;以^32P标记产物S1P测定细胞内SPK酶活性;用扩散盒技术观察高表达SPK^WT及SPK^ND对HGF诱导的内皮细胞迁移的影响。结果：野生型SPK基因表达可明显增强细胞内SPK的活性,并促进HGF诱导的内皮细胞迁移;而SPK负显性基因则显著抑制HGF诱导的内皮细胞迁移。结论：HGF通过SPK调控内皮细胞的迁移。

Sphingosine kinase regulates hepatocyte growth factor-induced migration of endothelial cells

Aim: To elucidate the effect of sphingosine kinase(SPK) on the hepatocye growth factor(HGF)-induced migration of endothelial cells.Methods: We constructed recombinant adenoviral vectors,which contain SPK gene and its mutant respectively.These adenoviral vectors were packaged and amplified in 293 cells.And intracellular SPK activity was assayed via measurement of ~()P radioisotope labeled S1P;the effect of SPK activation on HGF-induced migration of endothelial cell was observed by Transwell technique.Results: Adenoviral mediated expression of SPK gene increased in ECV 304 cells intracellular SPK activity,which in turn enhanced the HGF-induced migration.Whereas these activities were blocked by the dominant negative SPK gene. Conclusion: These findin show that SPK activation plays important roles in the regulation of HGF-induced migration of endothelial cells.