Increased focal adhesion kinase- and urokinase-type plasminogen activator receptor-associated cell signaling in endothelial cells exposed to asbestos.
Exposure of low-passage endothelial cells in culture to nonlethal amounts of asbestos, but not refractory ceramic fiber-1, increases cell motility and gene expression. These changes may be initiated by the fibers mimicking matrix proteins as ligands for receptors on the cell surface. In the present study, 1- to 3-hr exposures of endothelial cells to 5 mg/cm2 of chrysotile asbestos caused marked cell elongation and motility. However, little morphological change was seen when chrysotile was added to cells pretreated with either mannosamine to prevent assembly of glycophosphatidylinositol (GPI)-anchored receptors or with herbimycin A to inhibit tyrosine kinase activity. Affinity purification of GPI-anchored urokinase-type plasminogen activator receptor (uPAR) from chrysotile-exposed cells demonstrated that asbestos altered the profile of proteins and phosphoproteins complexed with this receptor. Tyrosine kinase activities in the complexes were also increased by asbestos. Immunoprecipitations with selective monoclonal antibodies demonstrated that both chrysotile and crocidolite asbestos increase kinase activities associated with p60 Src or p120 focal adhesion kinase (FAK). Further, chrysotile also changed the profile of proteins and phosphoproteins associated with FAK in intact cells. These data suggest that asbestos initiates endothelial cell phenotypic change through interactions with uPAR-containing complexes and that this change is mediated through tyrosine kinase cascades.
Barchowsky
A
A
Department of Pharmacology, Dartmouth Medical School, Hanover, NH 03755, USA. barchowsky@dartmouth.edu
Lannon
B M
BM
Elmore
L C
LC
Treadwell
M D
MD
D000818:Animals; D001194:Asbestos / Q000633:toxicity*; D002273:Carcinogens / Q000633:toxicity*; D002448:Cell Adhesion / Q000187:drug effects; D004727:Endothelium / Q000166:cytology / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D017395:Plasminogen Activator Inhibitor 1 / Q000502:physiology*; D017396:Plasminogen Activator Inhibitor 2 / Q000502:physiology*; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013552:Swine; D019061:src-Family Kinases / Q000096:biosynthesis / Q000235:genetics
D002273:Carcinogens; D017395:Plasminogen Activator Inhibitor 1; D017396:Plasminogen Activator Inhibitor 2; D001194:Asbestos; D011505:Protein-Tyrosine Kinases; D019061:src-Family Kinases