Thrombin Receptor Activating Peptide (TRAP) Stimulates Mitogenesis, c-fos and PDGF-A Gene Expression in Human Vascular Smooth Muscle Cells

Chryso Kanthou; Omar Benzakour; Geeta Patel; John Deadman; Vijay Vir Kakkar; Florea Lupu

doi:10.1055/s-0038-1649937

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1995; 74(05): 1340-1347
DOI: 10.1055/s-0038-1649937

Original Article

Vessel Wall

Schattauer GmbH Stuttgart

Thrombin Receptor Activating Peptide (TRAP) Stimulates Mitogenesis, c-fos and PDGF-A Gene Expression in Human Vascular Smooth Muscle Cells

Authors

Chryso Kanthou

The Vascular Biology Laboratory, Thrombosis Research Institute, Chelsea, London, UK
Omar Benzakour

The Vascular Biology Laboratory, Thrombosis Research Institute, Chelsea, London, UK
Geeta Patel

The Vascular Biology Laboratory, Thrombosis Research Institute, Chelsea, London, UK
John Deadman

The Vascular Biology Laboratory, Thrombosis Research Institute, Chelsea, London, UK
Vijay Vir Kakkar

The Vascular Biology Laboratory, Thrombosis Research Institute, Chelsea, London, UK
Florea Lupu

The Vascular Biology Laboratory, Thrombosis Research Institute, Chelsea, London, UK

Abstract

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Summary

The synthetic peptide SFLLRNPNDKYEPF, identical in sequence to the new amino-terminus of the thrombin receptor generated following cleavage by thrombin, acts as a thrombin receptor agonist/ activating peptide (TRAP). In this study, Northern blot analysis showed that cultured human vascular smooth muscle cells (HVSMC) express a thrombin receptor transcript. TRAP, in contrast to thrombin was shown to be a weak mitogen for HVSMC. A combination of TRAP and enzymatically-inactivated thrombin (PPACK-thrombin) which provides receptor occupancy, did not potentiate TRAP-induced mitogenesis, indicating that TRAP and PPACK-thrombin do not reproduce the mitogenic effect of enzymatically-active thrombin. Both thrombin and TRAP, induced the expression of c-fos and the PDGF-A gene in a pertussis toxin (PTX)-insensitive manner. Examination of thrombin and TRAP-treated cells by immunofluorescence staining followed by computer assisted image analysis revealed that thrombin and to a lesser extent TRAP induced PDGF-AA protein expression. Antibodies to PDGF-AA partially inhibited thrombin but not TRAP-induced mitogenesis in HVSMC. This study indicates that in addition to the common signalling pathways utilised by thrombin and TRAP, enzymatically-active thrombin activates other signalling pathways and hence TRAP does not mimic fully the biological effect of thrombin on HVSMC.

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References

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