Abstract
Previous studies from our laboratory have shown that during angiogenesis in vitro, rmMCP-7 (recombinant mouse mast cell protease-7) stimulates endothelial cell spreading and induces their penetration into the matrix. The ability of rmMCP-7 to induce angiogenesis in vivo was assessed in the present study using a directed in vivo angiogenesis assay (DIVAA™). Vessel invasion of the angioreactor was observed in the presence of rmMCP-7 but was not seen in the control. Since integrins are involved in endothelial cell migration, the relationship between rmMCP-7 and integrins during angiogenesis was investigated. Incubation with rmMCP-7 resulted in a reduction in the levels of integrin subunits αv and β1 on SVEC4-10 endothelial cells during angiogenesis in vitro. Furthermore, the degradation of integrin subunits occurs both through the direct action of rmMCP-7 and indirectly via the ubiquitin/proteasome system. Even in the presence of a proteasome inhibitor, incubation of endothelial cells with rmMCP-7 induced cell migration and tube formation as well as the beginning of loop formation. These data indicate that the direct degradation of the integrin subunits by rmMCP-7 is sufficient to initiate angiogenesis. The results demonstrate, for the first time, that mMCP-7 acts in angiogenesis through integrin degradation.
Highlights
Angiogenesis is a dynamic process that consists of the formation of new blood vessels from pre-existing ones which begins in the embryo and continues throughout life [1,2]
3.1. rmMCP-7 Protease Induces Endothelial cell Invasion of SVEC4-10 cells into Geltrex® increased in the presence of rmMCP-7
Since rmMCP-7 accelerates in vitro angiogenesis, it was of interest to determine whether the incubated in the presence of rmMCP-7 using a Secure-SealTM Hybridization Chamber Gasket attached to invasion of SVEC4-10 cells into Geltrex® increased in the presence of rmMCP-7
Summary
Angiogenesis is a dynamic process that consists of the formation of new blood vessels from pre-existing ones which begins in the embryo and continues throughout life [1,2]. Angiogenesis consists of three basic stages: the first, selection of some endothelial cells inside the capillary to begin angiogenic expansion. These cells react to the angiogenic factor VEGF-A (vascular endothelial growth factor) and initiate invasion and migration. Proliferation and migration of endothelial cells is inhibited, and the newly formed vessels fuse with other vessels, both new vessels and pre-existing vessels. This fusion stabilizes the newly formed vessels and forms a functional vascular network [6,7,8,9]
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