title: The role of the cytoskeleton in cellular adhesion molecule expression in tumor necrosis factor-stimulated endothelial cells.
abstract: Leukocyte infiltration is a hallmark of the atherosclerotic lesion. These cells are captured by cellular adhesion molecules (CAMs), including vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), platelet-endothelial cell adhesion molecule (PECAM), and E-selectin, on endothelial cells (EC). We examined the role of the actin cytoskeleton in tumor necrosis factor-alpha (TNF-alpha)-induced translocation of CAMs to the cell surface. Human aortic EC were grown on 96-well plates and an ELISA was used to assess surface expression of the CAMs. TNF-alpha increased VCAM-1, ICAM-1, and E-selectin by 4 h but had no affect on the expression of PECAM. A functioning actin cytoskeleton was important for VCAM-1 and ICAM-1 expression as both cytochalasin D, an actin filament disruptor, and jasplakinolide, an actin filament stabilizer, attenuated the expression of these CAMs. These compounds were ineffective in altering E-selectin surface expression. Myosin light chains are phosphorylated in response to TNF-alpha and this appears to be regulated by Rho kinase instead of myosin light chain kinase. However, the Rho kinase inhibitor, Y27632, had no affect on TNF-alpha-induced CAM expression. ML-7, a myosin light chain kinase inhibitor, had a modest inhibitory effect on the translocation of VCAM-1 but not on ICAM-1 or E-selectin. These data suggest that the surface expression of VCAM-1 and ICAM-1 is dependent on cycling of the actin cytoskeleton. Nevertheless, modulation of actin filaments via myosin light chain phosphorylation is not necessary. The regulation of E-selectin surface expression differs from that of the other CAMs.
authors.affiliation: Department of Surgery, Division of Vascular Surgery, University of Cincinnati Medical Center, 231 Albert Sabin Way, Cincinnati, Ohio 45267-0558, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: VandenBerg
authors.forename: Maria D
authors.initials: MD
authors.lastname: Reid
authors.forename: John D
authors.initials: JD
authors.lastname: Edwards
authors.forename: Harold W
authors.initials: HW
authors.lastname: Davis
mesh_terms: D000199:Actins / Q000378:metabolism; D000577:Amides / Q000494:pharmacology; D001381:Azepines / Q000494:pharmacology; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000378:metabolism*; D002460:Cell Line; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000502:physiology*; D047630:Depsipeptides / Q000494:pharmacology; D019040:E-Selectin / Q000032:analysis / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D018799:Intercellular Adhesion Molecule-1 / Q000032:analysis / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008856:Microscopy, Fluorescence; D018994:Myosin Light Chains / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism; D009281:Naphthalenes / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D019010:Vascular Cell Adhesion Molecule-1 / Q000032:analysis / Q000378:metabolism; D054460:rho-Associated Kinases
chemical_list: D000199:Actins; D000577:Amides; D001381:Azepines; D015815:Cell Adhesion Molecules; D047630:Depsipeptides; D019040:E-Selectin; D047908:Intracellular Signaling Peptides and Proteins; D018994:Myosin Light Chains; D009281:Naphthalenes; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011725:Pyridines; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; C057531:jasplakinolide; C070571:ML 7; D018799:Intercellular Adhesion Molecule-1; C108830:Y 27632; D015638:Cytochalasin D; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; D009219:Myosin-Light-Chain Kinase