Plasmin-clipped beta(2)-glycoprotein-I inhibits endothelial cell growth by down-regulating cyclin A, B and D1 and up-regulating p21 and p27.
beta(2)-Glycoprotein-I (beta(2)gpI), an abundant plasma glycoprotein, functions as a regulator of thrombosis. Previously, we demonstrated that plasmin-clipped beta(2)gpI (cbeta(2)gpI) exerts an anti-angiogenic effect on human umbilical vein endothelial cells (HUVEC). The present study was focused on the molecular background responsible for this phenomenon. cbeta(2)gpI strongly reduced HUVEC growth and proliferation as evidenced by the MTT and BrdU assay and delayed cell cycle progression arresting HUVEC in the S-and G2/M-phase. Western blot analysis indicated that cbeta(2)gpI inhibited cyclin A, B and D1, and enhanced p21 and p27 expression. Activity of p38 was down-regulated independently from the cbeta(2)gpI incubation time. Phosphorylation of ERK1/2 was not changed early (30 and 60 min) but became enhanced later (90 min, 4h). JNK activity was reduced rapidly after cbeta(2)gpI treatment but compared to controls, increased thereafter. Annexin II blockade prevented growth inhibition and cell cycle delay evoked by cbeta(2)gpI. We assume that cbeta(2)gpI's effects on HUVEC growth is mediated via cyclin A, B and D1 suppression, up-regulation of p21 and p27 and coupled to modifications of the mitogen-activated protein (MAP) kinase signalling pathway. cbeta(2)gpI may represent a potential endogenous angiogenesis-targeted compound, opening the possibility of a novel tool to treat cancer.
Beecken
Wolf-Dietrich C
WD
Department of Urology, J.W. Goethe-University, Frankfurt am Main, Germany.
Ringel
Eva Maria
EM
Babica
Jan
J
Oppermann
Elsie
E
Jonas
Dietger
D
Blaheta
Roman A
RA
D017306:Annexin A2 / Q000494:pharmacology; D002453:Cell Cycle; D002455:Cell Division / Q000502:physiology; D019925:Cyclin A / Q000235:genetics*; D019926:Cyclin B / Q000235:genetics*; D019938:Cyclin D1 / Q000235:genetics*; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics*; D015536:Down-Regulation; D004730:Endothelium, Vascular / Q000502:physiology*; D005341:Fibrinolysin / Q000502:physiology; D006801:Humans; D018919:Neovascularization, Physiologic / Q000502:physiology; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics*; D014471:Umbilical Veins / Q000166:cytology / Q000502:physiology; D015854:Up-Regulation; D053482:beta 2-Glycoprotein I / Q000302:isolation & purification / Q000378:metabolism* / Q000502:physiology
D017306:Annexin A2; C493956:CDKN1A protein, human; D019925:Cyclin A; D019926:Cyclin B; D050759:Cyclin-Dependent Kinase Inhibitor p21; D018809:Proliferating Cell Nuclear Antigen; D053482:beta 2-Glycoprotein I; C413374:p27 antigen; D019938:Cyclin D1; D005341:Fibrinolysin