Regulation of Plasminogen Activator Inhibitor-1 mRNA in Human Endothelial Cells

E A van den Berg; E D Sprengers; M Jaye; W Burgess; T Maciag; V W M van Hinsbergh

doi:10.1055/s-0038-1647636

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1988; 60(01): 063-067
DOI: 10.1055/s-0038-1647636

Original Article

Schattauer GmbH Stuttgart

Regulation of Plasminogen Activator Inhibitor-1 mRNA in Human Endothelial Cells

E A van den Berg

¹The Gaubius Institute TNO, Leiden, The Netherlands

,

E D Sprengers

¹The Gaubius Institute TNO, Leiden, The Netherlands

,

M Jaye

²The Rorer Biotechnology Inc., Springfield, VA, USA

,

W Burgess

³The Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross, Rockville, MD, USA

,

T Maciag

³The Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross, Rockville, MD, USA

,

V W M van Hinsbergh

¹The Gaubius Institute TNO, Leiden, The Netherlands

› Author Affiliations

Abstract

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Summary

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.

Key words

Endothelial cells - Plasminogen-activator inhibitor - Messenger RNA - Tumor necrosis factor - Endotoxin

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References

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