Regulation of plasminogen activator inhibitor-1 mRNA in human endothelial cells
article
The plasminogen activator inhibitor (PAI-1) from endothelial cells is a potentially important regulator of plasminogen activator activity. Cultured human endothelial cells increase their PAI-1 production upon stimulation with LPS and TNF, agents that are known to cause an increase in PAI-1 levels in vivo. We isolated a PAI-cDNA probe, and by RNA hybridization analysis studied the regulation of PAI-1 mRNA synthesis in human endothelial artery cells. Freshly isolated endothelial cells do not contain detectable amounts of PAI-1 mRNA, but after adherence and incubation for 18 h in growth medium produce considerable amounts of PAI-1 activity and contain PAI-1 mRNA levels comparable to those found in subcultured cells. When subcultured endothelial cells are incubated for 6 h with LPS or TNF, both species of PAI-1 mRNA increase 10 to 20 fold, while PAI-1 activity in the growth medium increases only 1.5 to 2 fold. Stimulation of endothelial cells in the presence of cycloheximide (CHX) results in superinduction of mainly the 3.0 kb PAI-1 mRNA. The 3' end of this mRNA contains a 60 bp AT-rich sequence, that resembles 3' sequences present in a number of other genes superinducible with CHX. Chemicals/CAS: plasminogen activator inhibitor, 105844-41-5; RNA, 63231-63-0; Culture Media; Cycloheximide, 66-81-9; DNA Probes; Glycoproteins; Lipopolysaccharides; Plasminogen Inactivators; RNA, 63231-63-0; RNA, Messenger; Tumor Necrosis Factor
Topics
endotoxinmessenger rnaplasminogen activator inhibitorrnatumor necrosis factoradultbiological modelcytochemistrydna probeendothelium cellfemalehuman cellhybridizationmalemethodologyrna fingerprintingtissue cultureBase SequenceBlotting, NorthernCells, CulturedCloning, MolecularCulture MediaCycloheximideDNA ProbesElectrophoresis, Agar GelEndothelium, VascularGene Expression RegulationGlycoproteinsHumanLipopolysaccharidesNucleic Acid HybridizationPlasminogen InactivatorsRNARNA, MessengerSupport, Non-U.S. Gov'tTumor Necrosis Factor
TNO Identifier
230636
ISSN
03406245
Source
Thrombosis and Haemostasis, 60(1), pp. 63-67.
Pages
63-67
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