title: Hypoxia induces PECAM-1 phosphorylation and transendothelial migration of monocytes.
abstract: Polymorphonuclear neutrophils (PMN) adhere to the vascular endothelium under hypoxic conditions, causing microvascular injury. The molecular mechanism of hypoxia-induced adhesion of PMN to and diapedesis through the vascular endothelium is poorly understood. We examined the effects of hypoxia on the transendothelial migration of monocytes. Exposure of human umbilical vein endothelial cells (HUVEC) cultured in Transwell chambers under low oxygen tension (3% O2 compared with 21% O2) resulted in an increased rate of migration of both monocyte-like HL-60 cells and human peripheral blood monocytes. Migration was inhibited by addition of an antibody to platelet endothelial cell adhesion molecule-1 (PECAM-1), a protein kinase C (PKC) inhibitor, or a platelet-activating factor (PAF)-receptor antagonist. In HUVEC, hypoxic conditions (1, 3, 5, and 14% O2) increased the phosphorylation of PECAM-1. The extent of phosphorylation of PECAM-1 was inversely related to the concentration of oxygen to which HUVEC were exposed. Hypoxia-induced phosphorylation of PECAM-1 was inhibited by either a PKC inhibitor or a PAF-receptor antagonist, indicating the involvement of hypoxia-induced release of PAF in both PKC activation and the concomitant phosphorylation of PECAM-1. These results were substantiated by the findings that treatment of HUVEC with 100 nM PAF under normoxic conditions augmented 11.8-fold the phosphorylation of PECAM-1 and twofold increase in the transendothelial migration of monocyte-like HL-60 cells. We conclude that PAF, produced by cultured endothelial cells in response to hypoxia, acts in an autocrine fashion to activate PKC, causing PECAM-1 phosphorylation and thus the transendothelial migration of monocytes.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern California School of Medicine, Los Angeles 90033, USA.
authors.forename: V K
authors.initials: VK
authors.lastname: Kalra
authors.forename: Y
authors.initials: Y
authors.lastname: Shen
authors.forename: C
authors.initials: C
authors.lastname: Sultana
authors.forename: V
authors.initials: V
authors.lastname: Rattan
mesh_terms: D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D066246:ErbB Receptors / Q000037:antagonists & inhibitors; D018922:HL-60 Cells; D006801:Humans; D000860:Hypoxia / Q000378:metabolism*; D009000:Monocytes / Q000502:physiology*; D010100:Oxygen / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010972:Platelet Activating Factor / Q000494:pharmacology; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism* / Q000502:physiology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D010972:Platelet Activating Factor; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D066246:ErbB Receptors; D011493:Protein Kinase C; D010100:Oxygen