Inhibition of angiogenic tubule formation and induction of apoptosis in human endothelial cells by the selective cyclooxygenase-2 inhibitor 5-bromo-2-(4-fluorophenyl)-3-(methylsulfonyl) thiophene (DuP-697).
There are indications that inhibitors of the cyclooxygenase-2 (COX-2) enzyme may cause inhibition of angiogenesis, proliferation of endothelial cells and induce apoptosis in cell systems. The concentrations of inhibitors required for such effects are however much higher than those needed to inhibit COX-2, suggesting that the latter may not be involved in these actions of the drugs. We have however generated data that strongly indicates a critical role for COX-2 suppression in the inhibition of angiogenesis and induction of apoptosis in human cultured umbilical vein endothelial cells (HUVECs) by the selective cyclooxygenase-2 (COX-2) inhibitor 5-bromo-2-(4-fluorophenyl)-3-(methylsulfonyl) thiophene (DuP-697). DuP-697 concentration-dependently inhibited prostaglandin E(2) (PGE(2)) production by HUVECs and at its known IC(50) for COX-2 inhibition of 10 nM inhibited basal and vascular endothelial cell growth factor (VEGF)-induced PGE(2) production by 80% and 85% respectively. DuP-697 also induced apoptosis as shown by FACs analysis, an increase in chromatin condensation and DNA laddering in HUVECS treated with the drug. Moreover, these effects were reversed by PGE(2) and by VEGF. In parallel studies, DuP-697 induced caspases 3, 8 and 9, with the caspase-3 specific inhibitor N-Acetyl-Asp-Glu-Val-Asp-al (DEVD-CHO) blocking the induction of apoptosis. Capillary-like tubule formation by HUVECs cultured on Matrigel was inhibited by DuP-697 and this inhibition was prevented by PGE(2) but not by DEVD-CHO. These results indicate that the induction of apoptosis and inhibition of tubule formation by DuP-697 involves the inhibition of COX-2 and that whereas the induction of apoptosis is caspase-dependent, the inhibition of tubule formation occurs through a caspase-independent mechanism.
Churchman
Adrian
A
School of Life Sciences, University of Hertfordshire, College Lane, Hatfield, AL10 9AB, UK.
Baydoun
Anwar R
AR
Hoffman
Richard
R
D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D061945:Caspase Inhibitors; D002478:Cells, Cultured; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects; D003094:Collagen / Q000737:chemistry; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D051546:Cyclooxygenase 2 / Q000378:metabolism; D052246:Cyclooxygenase 2 Inhibitors / Q000494:pharmacology*; D053938:DNA Fragmentation / Q000187:drug effects; D015237:Dinoprost / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology; D007797:Laminin / Q000737:chemistry; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D009842:Oligopeptides / Q000494:pharmacology; D011509:Proteoglycans / Q000737:chemistry; D013876:Thiophenes / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology
C078129:6-keto-prostaglandin F2alpha; D061945:Caspase Inhibitors; D016895:Culture Media, Serum-Free; D052246:Cyclooxygenase 2 Inhibitors; D004338:Drug Combinations; D007797:Laminin; D009842:Oligopeptides; D011509:Proteoglycans; D013876:Thiophenes; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C108637:aspartyl-glutamyl-valyl-aspartal; C059485:matrigel; C064874:DuP 697; D003094:Collagen; D015237:Dinoprost; D051546:Cyclooxygenase 2; D053148:Caspase 3; D053181:Caspase 8; D015232:Dinoprostone; D007213:Indomethacin