Regulation of plasminogen activator inhibitor-1 mRNA in human endothelial cells.
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-1 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.
van den Berg
E A
EA
Gaubius Institute TNO, Leiden, The Netherlands.
Sprengers
E D
ED
Jaye
M
M
Burgess
W
W
Maciag
T
T
van Hinsbergh
V W
VW
D001483:Base Sequence; D015152:Blotting, Northern; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003470:Culture Media; D003513:Cycloheximide / Q000494:pharmacology; D015342:DNA Probes; D004587:Electrophoresis, Agar Gel; D004730:Endothelium, Vascular / Q000166:cytology*; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D009693:Nucleic Acid Hybridization; D015849:Plasminogen Inactivators; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D003470:Culture Media; D015342:DNA Probes; D006023:Glycoproteins; D008070:Lipopolysaccharides; D015849:Plasminogen Inactivators; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D012313:RNA; D003513:Cycloheximide