Identification And Application Of Sphingosine-1-phosphate Receptor Of Human Umbilical Vein Endothelial Cells

Objective: To identify the expression of sphingosine-1-phosphate receptor(S1PR) subtypes, c-myc and his tag proteins of human umbilical vein endothelial fusion cell line, EA.hy926 and human umbilical vein endothelial cells(HUVEC), CRL-1730 for studying the function of apolipoprotein M(ApoM)-S1 P axis.Methods: Two kinds of cells were cultured to reach the density of 60%-70% in vitro.Immunofluorescence technique was employed to investigate the expressions of coagulation factor Ⅷ(FⅧ), ApoM, S1PR1~S1PR5, c-myc and his tag proteins.Results:(1) Two kinds of cells both expressed FⅧ and ApoM. FⅧ presented scattered particle distribution in CRL-1730, while uniform distribution in EA.hy926. However, ApoM was strongly expressed and widely distributed in cytoplasm of two kinds of cells.(2) S1PR1-3 could be detected on their membrane other than S1PR4 and S1PR5. S1PR1 was highly expressed but S1PR2 and S1PR3 were in a low level expression.(3) Two kinds of cells both expressed c-myc and his tag proteins in cytoplasm.Conclusion: Two kinds of cells have the properties of endothelial cells and can express FⅧ, ApoM, c-myc and his tag proteins; It is not suitable for choosing c-myc and/or his tag–conjugated recombinant ApoM to research the fuction of ApoM-S1 P axis with these two kinds of cells.