title: Actions of the Kunitz-type serine protease inhibitor Amblyomin-X on VEGF-A-induced angiogenesis.
abstract: Amblyomin-X is a Kunitz-type serine protease inhibitor (Kunitz-type SPI) designed from the cDNA library of the Amblyomma cajennense tick, which displays in vivo anti-tumor activities. Here, the mechanisms of actions of Amblyomin-X in vascular endothelial growth factor A (VEGF-A)-induced angiogenesis were characterized. Topical application of Amblyomin-X (10 or 100 ng/10 μl; each 48 h) inhibited VEGF-A-induced (10 ng/10 μl; each 48 h) angiogenesis in the dorsal subcutaneous tissue in male Swiss mice. Moreover, similar effect was observed in the VEGF-A-induced angiogenesis in the chicken chorioallantoic membrane (CAM). Additional in vitro assays in t-End cells showed that Amblyomin-X treatment delayed the cell cycle, by maintaining them in G0/G1 phase, and inhibited cell proliferation and adhesion, tube formation and membrane expression of the adhesion molecule platelet-endothelial cell adhesion molecule-1 (PECAM-1), regardless of mRNA synthesis. Together, results herein reveal the role of Kunitz-type SPI on in vivo VEGF-A-induced angiogenesis, by exerting modulatory actions on endothelial cell proliferation and adhesion, especially on membrane expression of PECAM-1. These data provide further mechanisms of actions of Kunitz-type SPI, corroborating their relevance as scientific tools in the design of therapeutic molecules.
authors.affiliation: Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo, SP, Brazil.
authors.forename: C C
authors.initials: CC
authors.lastname: Drewes
authors.forename: R Y S
authors.initials: RY
authors.lastname: Dias
authors.forename: C B
authors.initials: CB
authors.lastname: Hebeda
authors.forename: S M
authors.initials: SM
authors.lastname: Simons
authors.forename: S A
authors.initials: SA
authors.lastname: Barreto
authors.forename: J M
authors.initials: JM
authors.lastname: Ferreira
authors.forename: A M
authors.initials: AM
authors.lastname: Chudzinski-Tavassi
authors.forename: S H P
authors.initials: SH
authors.lastname: Farsky
mesh_terms: D020533:Angiogenesis Inhibitors / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D060829:Arthropod Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002461:Cell Line, Transformed; D049109:Cell Proliferation / Q000187:drug effects; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000187:drug effects / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D065427:Factor Xa Inhibitors; D026863:Ixodidae / Q000378:metabolism; D008297:Male; D051379:Mice; D009389:Neovascularization, Pathologic / Q000188:drug therapy*; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D016192:Resting Phase, Cell Cycle / Q000187:drug effects; D012471:Salivary Proteins and Peptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D040521:Subcutaneous Tissue / Q000187:drug effects / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: C559369:Amblyomin-X protein, Amblyomma cajennense; D020533:Angiogenesis Inhibitors; D060829:Arthropod Proteins; D065427:Factor Xa Inhibitors; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides; D015842:Serine Proteinase Inhibitors; D042461:Vascular Endothelial Growth Factor A