Identification and application of sphingosine-1-phosphate receptor of human umbilical vein endothelial cells
10.11958/20160032
- VernacularTitle:人脐静脉内皮细胞1-磷酸鞘氨醇受体鉴定及其应用探讨
- Author:
Yun LIANG
;
Guanghua LUO
;
Hong LIU
;
Miaomei YU
;
Min WANG
;
Hongyao LIU
;
Bo HUANG
;
Ning XU
;
Xiaoying ZHANG
- Publication Type:Journal Article
- Keywords:
endothelial cells;
sphingosine-1-phosphate receptor;
apolipoprotein M;
coagulation factorⅧ;
proto-onco-gene proteins c-myc;
His tag protein
- From:
Tianjin Medical Journal
2016;44(6):662-664,655
- CountryChina
- Language:Chinese
-
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.