The use of suppression subtractive hybridization for the study of SDF-1alpha induced gene-expression in human endothelial cells.
Stromal cell-derived factor-1 (SDF-1), the only ligand of the CXCR4 receptor, is mainly known as a chemotactic factor for hematopoietic progenitor cells. However, studies of knock-out mice have shown malformation of different organ-systems suggesting that SDF-1 may have a role in angiogenesis and cardiac and cerebral development. However, the underlying mechanisms of its action are largely unknown. Therefore, we performed suppression subtractive hybridization (SSH) in order to identify genes that are differentially expressed after stimulation of human arterial endothelial cells (HUAEC) with SDF-1. Using SSH we found ten genes, with varied functions, whose mRNA expression is induced by SDF-1alpha in HUAEC. We show that SSH is a reliable method for identifying differentially expressed genes and that SDF-1alpha may have more functions than previously reported.
Medizinische Universitäts-Poliklinik Bonn, Wilhelmstr 35-37, Bonn 53111, Germany.
Thomas
T
Neuhaus
Christoph
C
Lutz
Sebastian
S
Stier
Gudrun
G
Totzke
Elisabeth
E
Gruenewald
Stefan
S
Fronhoffs
Agapios
A
Sachinidis
Hans
H
Vetter
Yon D
YD
Ko
D000818:Animals; D002478:Cells, Cultured; D054377:Chemokine CXCL12; D019743:Chemokines, CXC / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000187:drug effects*; D020869:Gene Expression Profiling; D006801:Humans; D051379:Mice; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000235:genetics; D019718:Receptors, CXCR4 / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; C515256:Cxcl12 protein, mouse; D012333:RNA, Messenger; D019718:Receptors, CXCR4