Antiangiogenic property of human thrombin.
Using protein chromatography, we purified and identified human prothrombin from human plasma as antiangiogenic. Prothrombin significantly inhibited endothelial cell tube formation in vitro at 10 microg/ml. Importantly, it also inhibited bFGF-induced angiogenesis in Matrigel-plug assays performed in mice. The proteolytic activity of thrombin appeared to be critical for the antiangiogenic activity of prothrombin. For example, thrombin exhibited inhibitory effects on endothelial cell tube formation in vitro at 10 U/ml. Addition of lepirudin, a specific inhibitor of thrombin, completely blocked prothrombin's and thrombin's antiangiogenic effects in vitro. We also assessed the importance of thrombin receptors in angiogenesis. Using small peptides that activate different protease-activated receptors (PARs), we showed that activation of PAR-1 led to inhibition of endothelial cell tube formation in vitro and bFGF-induced angiogenesis in vivo. Collectively, our data suggest that thrombin's proteolytic activity can be antiangiogenic.
Chan
Barden
B
Division of Nephrology, Center for Study of the Tumor Microenvironment, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA. vsukhatm@caregroup.harvard.com
Merchan
Jaime R
JR
Kale
Sujata
S
Sukhatme
Vikas P
VP
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D002455:Cell Division; D002478:Cells, Cultured; D002845:Chromatography; D002852:Chromatography, Ion Exchange; D003094:Collagen / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D009389:Neovascularization, Pathologic; D010455:Peptides / Q000737:chemistry; D010703:Phorbol Esters / Q000378:metabolism; D011509:Proteoglycans / Q000494:pharmacology; D011516:Prothrombin / Q000378:metabolism; D044463:Receptor, PAR-1 / Q000378:metabolism; D044462:Receptors, Proteinase-Activated / Q000378:metabolism; D013917:Thrombin / Q000378:metabolism / Q000494:pharmacology* / Q000502:physiology*; D014471:Umbilical Veins / Q000166:cytology
D020533:Angiogenesis Inhibitors; D004338:Drug Combinations; D007797:Laminin; D010455:Peptides; D010703:Phorbol Esters; D011509:Proteoglycans; D044463:Receptor, PAR-1; D044462:Receptors, Proteinase-Activated; C059485:matrigel; D011516:Prothrombin; D003094:Collagen; D010450:Endopeptidases; D013917:Thrombin