Post-transcriptional control of Amblyomin-X on secretion of vascular endothelial growth factor and expression of adhesion molecules in endothelial cells.
Angiogenesis is a pivotal process of homeostasis and tissue repair, but it also favours neovascularisation syndromes and cancer nutrition. The chemical mediation of angiogenesis is complex, involving a balance between serine proteases and their inhibitors. We addressed the mechanisms of action of a Kunitz serine protease inhibitor (KPI) on spontaneous angiogenesis, using Amblyomin-X, a KPI designed from the cDNA library of the Amblyomma cajennense tick. Amblyomin-X treatment (10-1000 ng/10 μL; each 48 h; 3 times) reduced the number of vessels in the subcutaneous dorsal tissue of male Swiss mice, as measured by intravital microscopy, haematoxylin-eosin staining, and PECAM-1 immunofluorescence labeling. Incubation of Amblyomin-X with t-End endothelial cells, a murine endothelial microvascular lineage, did not alter cell proliferation, cell-cycle phases, necrosis and apoptosis, and the production of nitric oxide and prostaglandin E2. Nevertheless, Amblyomin-X treatment reduced t-End migration and adhesion to Matrigel(®), and inhibited the VEGF-A secretion and VCAM-1 and β3 integrin expressions by posttranscriptional pathways. Together, data herein outline novel posttranscriptional mechanisms of KPIs on endothelial cells during angiogenesis and point out the possible application of Amblyomin-X as a local inhibitor to undesired neovascularisation process.
Laboratory of Experimental Toxicology, Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, Ave. Prof. Lineu Prestes 580, SP, 05503-900, São Paulo, Brazil.
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Drewes
Laboratory of Experimental Toxicology, Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, Ave. Prof. Lineu Prestes 580, SP, 05503-900, São Paulo, Brazil.
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Dias
Biochemistry and Biophysics Laboratory, Butantan Institute, Ave. Vital Brasil 1500, SP, 05503-900, São Paulo, Brazil.
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Branco
Laboratory of Biochemistry, Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, Ave. Prof. Lineu Prestes 580, SP, 05503-900, São Paulo, Brazil.
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Cavalcante
Biochemistry and Biophysics Laboratory, Butantan Institute, Ave. Vital Brasil 1500, SP, 05503-900, São Paulo, Brazil.
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Souza
Laboratory of Biochemistry, Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, Ave. Prof. Lineu Prestes 580, SP, 05503-900, São Paulo, Brazil.
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Abdalla
Biochemistry and Biophysics Laboratory, Butantan Institute, Ave. Vital Brasil 1500, SP, 05503-900, São Paulo, Brazil.
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Chudzinski-Tavassi
Laboratory of Experimental Toxicology, Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, Ave. Prof. Lineu Prestes 580, SP, 05503-900, São Paulo, Brazil. Electronic address: sfarsky@usp.br.
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Farsky
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D060829:Arthropod Proteins; D002448:Cell Adhesion / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015232:Dinoprostone / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation; D015723:Gene Library; D008297:Male; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects; D009569:Nitric Oxide / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012471:Salivary Proteins and Peptides / Q000235:genetics / Q000494:pharmacology*; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
C559369:Amblyomin-X protein, Amblyomma cajennense; D060829:Arthropod Proteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D012333:RNA, Messenger; D012471:Salivary Proteins and Peptides; D019010:Vascular Cell Adhesion Molecule-1; D042461:Vascular Endothelial Growth Factor A; D009569:Nitric Oxide; D015232:Dinoprostone
Amblyomma cajanense; Angiogenesis; Dorsal chamber; Post-transcriptional control; β(3) integrin and VCAM-1