DOI: 10.11958/20160032
Identification and application of sphingosine-1-phosphate receptor of human umbilical vein endothelial cells
LIANG Yun
LUO Guanghua
LIU Hong
YU Miaomei
WANG Min
LIU Hongyao
HUANG Bo
XU Ning
ZHANG Xiaoying
Abstract：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)-S1P axis. Methods Two kinds of cells (EA. hy926 and CRL-1730) were cultured to reach the density of 60%-70% in vitro. Immunofluorescence technique was em?ployed to investigate the expressions of coagulation factorⅧ(FⅧ), ApoM, S1PR1-S1PR5, C-myc and His tag proteins. Re?sults (1) Two kinds of cells both expressed FⅧand ApoM. FⅧpresented 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 can 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 study the fuction of ApoM-S1P axis with these two kinds of cells.
Keywords：endothelial cellssphingosine-1-phosphate receptorapolipoprotein Mcoagulation factorⅧproto-onco-gene proteins c-mycHis tag protein
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 662-664,655 )