Elastin-derived peptides enhance angiogenesis by promoting endothelial cell migration and tubulogenesis through upregulation of MT1-MMP
Open Access
- 15 January 2005
- journal article
- research article
- Published by The Company of Biologists in Journal of Cell Science
- Vol. 118 (2), 343-356
- https://doi.org/10.1242/jcs.01613
Abstract
Elastin-derived peptides display a wide range of biological activities in a number of normal and transformed cells but their involvement in angiogenesis has not been reported. In the present study, we show that κ-elastin and VGVAPG hexapeptide elastin motif accelerated angiogenesis in the chick chorio-allantoic membrane in an in vivo model. They also stimulated pseudotube formation from human vascular and microvascular endothelial cells in the matrigel and collagen models as well as cell migration in an in vitro wound healing assay. Confocal and scanning electron microscopy analyses revealed the main reorganization of actin filaments mediated by elastin-derived peptides and changes in cell shape that correlated with a decrease of the cell form factor determined by computerized image analysis. Such elastin-derived peptide effects were attributed to upregulation of proMT1-MMP and proMMP-2 expression and activation at both the mRNA and protein levels. Batimastat, an inhibitor of furin convertase and TIMP-2, but not TIMP-1, totally abolished the influence of elastin-derived peptides (EDPs) on cell migration and tubulogenesis, thus favoring the involvement of MT1-MMP in such processes. To assess its contribution to EDP-mediated angiogenesis further, we used a small interfering RNA (siRNA) approach for specifically silencing MT1-MMP in human microvascular endothelial cells. Four sets of 21 bp siRNA duplexes targeting MT1-MMP mRNA were synthesized by in vitro transcription. Two of them proved to inhibit MT1-MMP expression efficiently but did not affect MT2-, MT3- and MT5-MMP expression. Seventy-two hours after transfection with 25 nM siRNAs EDP-induced MT1-MMP expression at the mRNA and protein levels was decreased fourfold. In parallel, proMMP-2 activation was inhibited. A scrambled siRNA, used as a negative control, had no effect. Finally, the effect of elastin peptides on pseudotube formation in MT1-MMP-siRNA transfected cells was totally abolished. These data emphasise the crucial role of MT1-MMP in the elastin-induced angiogenic phenotype of endothelial cells.Keywords
This publication has 79 references indexed in Scilit:
- Matrix metalloproteinases and matrikines in angiogenesisCritical Reviews in Oncology/Hematology, 2004
- Membrane type-1 matrix metalloproteinase (MT1-MMP) processing of pro-αv integrin regulates cross-talk between αvβ3 and α2β1 integrinsin breast carcinoma cellsExperimental Cell Research, 2003
- Matrix-dependent Proteolysis of Surface Transglutaminase by Membrane-type Metalloproteinase Regulates Cancer Cell Adhesion and LocomotionOnline Journal of Public Health Informatics, 2001
- Conformational Dependence of Collagenase (Matrix Metalloproteinase-1) Up-regulation by Elastin Peptides in Cultured FibroblastsOnline Journal of Public Health Informatics, 2001
- Angiogenesis in cancer and other diseasesNature, 2000
- Mechanisms of angiogenesis and arteriogenesisNature Medicine, 2000
- Regulation of matrix metalloproteinase-2(gelatinase A, MMP-2), membrane-type matrixmetalloproteinase-1 (MT1-MMP) and tissue inhibitorof metalloproteinases-2 (TIMP-2) expression byelastin-derived peptides in human HT-1080 fibrosarcoma cell lineClinical & Experimental Metastasis, 1998
- HMEC-1: Establishment of an Immortalized Human Microvascular Endothelial Cell LineJournal of Investigative Dermatology, 1992
- Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extractionAnalytical Biochemistry, 1987
- Practical MorphometryPublished by Springer Science and Business Media LLC ,1983