Expression and activity of urokinase and its receptor in endothelial and pulmonary epithelial cells exposed to asbestos.
An elongated endothelial cell phenotype, which demonstrated increased ICAM-1-dependent neutrophil adherence, was induced when these cells were exposed to noncytotoxic concentrations of asbestos (Treadwell et al., Toxicol. Appl. Pharmacol. 139, 62-70, 1996). The present study examined mechanisms underlying this phenotypic change by investigating the effects of asbestos on transcription factor activation and expression of urokinase-type plasminogen activator (uPA) and its receptor uPAR. In situ zymography was used to compare the effects of these fibers on the activity of uPA. Cultures incubated with chrysotile or crocidolite asbestos, but not refractory ceramic fiber 1 (RCF-1), demonstrate localized cleavage of plasminogen, which was inhibited by amiloride. Immunocytochemistry showed that chrysotile-stimulated uPA activity was associated with a time-dependent augmentation of uPAR protein levels. RT-PCR analysis was used to investigate molecular mechanisms for these increases. Chrysotile asbestos, but not RCF-1, increased endothelial cell uPA message, relative to changes in beta-actin mRNA. This response to asbestos was not limited to endothelial cells, since both uPA and uPAR mRNA levels increase in human bronchial epithelial BEAS-2B cells exposed to chrysotile fibers. Finally, both types of asbestos, but not RCF-1, increased nuclear levels of nuclear factor-kappaB (NF-kappa B), a transcription factor common to increased expression of ICAM-1 and uPA. These data demonstrate that asbestos caused fiber-specific activation of endothelial and pulmonary epithelial cells, resulting in phenotypes capable of facilitating tissue remodeling.
Barchowsky
A
A
Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, New Hampshire. barchowsky@dartmouth.edu
Roussel
R R
RR
Krieser
R J
RJ
Mossman
B T
BT
Treadwell
M D
MD
D000818:Animals; D001194:Asbestos / Q000633:toxicity*; D017632:Asbestos, Serpentine / Q000633:toxicity; D001980:Bronchi / Q000166:cytology / Q000187:drug effects; D002273:Carcinogens / Q000633:toxicity*; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000201:enzymology; D004847:Epithelial Cells / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008168:Lung / Q000166:cytology / Q000187:drug effects; D016328:NF-kappa B / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000096:biosynthesis*; D055293:Receptors, Urokinase Plasminogen Activator; D013552:Swine; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis*
D017632:Asbestos, Serpentine; D002273:Carcinogens; D016328:NF-kappa B; C524898:PLAUR protein, human; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D001194:Asbestos; D010447:Peptide Hydrolases; D014568:Urokinase-Type Plasminogen Activator