title: Src and phosphatidylinositol 3-kinase mediate soluble E-selectin-induced angiogenesis.
abstract: Angiogenesis plays an important role in a variety of pathophysiologic processes, including tumor growth and rheumatoid arthritis. We have previously shown that soluble E-selectin (sE-selectin) is an important angiogenic mediator. However, the mechanism by which sE-selectin mediates angiogenesis is still unknown. In this study, we show that sE-selectin is a potent mediator of human dermal microvascular endothelial cell (HMVEC) chemotaxis, which is predominantly mediated through the Src and the phosphatidylinositiol 3-kinase (PI3K) pathways. Further, sE-selectin induced a 2.2-fold increase in HMVEC tube formation in the Matrigel in vitro assay. HMVECs pretreated with the Src inhibitor (PP2) and the PI3K inhibitor (LY294002) or transfected with Src antisense oligonucleotides or Akt dominant-negative mutants significantly inhibited sE-selectin-mediated HMVEC tube formation. In contrast, HMVECs transfected with an extracellular signal-related kinase 1/2 (ERK1/2) mutant or pretreated with the mitogen-activated protein kinase (MAPK) inhibitor PD98059 failed to show sE-selectin-mediated HMVEC tube formation. Similarly, in the Matrigel-plug in vivo assay, sE-selectin induced a 2.2-fold increase in blood vessel formation, which was significantly inhibited by PP2 and LY294002 but not by PD98059. sE-selectin induced a marked increase in Src, ERK1/2, and PI3K phosphorylation. PI3K and ERK1/2 phosphorylation was significantly inhibited by PP2, thereby suggesting that both of these pathways may be activated via Src kinase. Even though the ERK1/2 pathway was activated by sE-selectin in HMVECs, it seems not to be essential for sE-selectin-mediated angiogenesis. Taken together, our data clearly show that sE-selectin-induced angiogenesis is predominantly mediated through the Src-PI3K pathway.
authors.affiliation: Veterans Administration, Lakeside Division, Chicago Health Care System, Chicago, IL, USA.
authors.forename: Pawan
authors.initials: P
authors.lastname: Kumar
authors.forename: Mohammad A
authors.initials: MA
authors.lastname: Amin
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Harlow
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Polverini
authors.forename: Alisa E
authors.initials: AE
authors.lastname: Koch
mesh_terms: D002478:Cells, Cultured; D002633:Chemotaxis / Q000187:drug effects / Q000502:physiology; D002867:Chromones / Q000494:pharmacology; D003094:Collagen; D004338:Drug Combinations; D019040:E-Selectin / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007797:Laminin; D008833:Microcirculation / Q000187:drug effects / Q000502:physiology; D009025:Morpholines / Q000494:pharmacology; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011509:Proteoglycans; D011994:Recombinant Proteins / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D012867:Skin / Q000098:blood supply; D014471:Umbilical Veins; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: D002867:Chromones; D004338:Drug Combinations; D019040:E-Selectin; D004791:Enzyme Inhibitors; D007797:Laminin; D009025:Morpholines; D011509:Proteoglycans; D011994:Recombinant Proteins; C059485:matrigel; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D003094:Collagen; D019869:Phosphatidylinositol 3-Kinases; D019061:src-Family Kinases