Alternagin-C binding to α2β1 integrin controls matrix metalloprotease-9 and matrix metalloprotease-2 in breast tumor cells and endothelial cells

Abstract

BackgroundMatrix metalloproteinases (MMPs) are key players in tumor progression, helping tumor cells to modify their microenvironment, which allows cell migration to secondary sites. The role of integrins, adhesion receptors that connect cells to the extracellular matrix, in MMP expression and activity has been previously suggested. However, the mechanisms by which integrins control MMP expression are not completely understood. Particularly, the role of α2β1 integrin, one of the major collagen I receptors, in MMP activity and expression has not been studied. Alternagin-C (ALT-C), a glutamate-cysteine-aspartate-disintegrin from Bothrops alternatus venom, has high affinity for an α2β1 integrin. Herein, we used ALT-C as a α2β1 integrin ligand to study the effect of ALT-C on MMP-9 and MMP-2 expression as well as on tumor cells, fibroblats and endothelial cell migration.MethodsALT-C was purified by two steps of gel filtration followed by anion exchange chromatography. The α2β1 integrin binding properties of ALT-C, its dissociation constant (Kd) relative to this integrin and to collagen I (Col I) were determined by surface plasmon resonance. The effects of ALT-C (10, 40, 100 and 1000 nM) in migration assays were studied using three human cell lines: human fibroblasts, breast tumor cell line MDA-MB-231, and microvascular endothelial cells HMEC-1, considering cells found in the tumor microenvironment. ALT-C effects on MMP-9 and MMP-2 expression and activity were analyzed by quantitative PCR and gelatin zymography, respectively. Focal adhesion kinase activation was determined by western blotting.ResultsOur data demonstrate that ALT-C, after binding to α2β1 integrin, acts by two distinct mechanisms against tumor progression, depending on the cell type: in tumor cells, ALT-C decreases MMP-9 and MMP-2 contents and activity, but increases focal adhesion kinase phosphorylation and transmigration; and in endothelial cells, ALT-C inhibits MMP-2, which is necessary for tumor angiogenesis. ALT-C also upregulates c-Myc mRNA level, which is related to tumor suppression.ConclusionThese results demonstrate that α2β1 integrin controls MMP expression and reveal this integrin as a target for the development of antiangiogenic and antimetastatic therapies.Electronic supplementary materialThe online version of this article (10.1186/s40409-018-0150-2) contains supplementary material, which is available to authorized users.

Cell migration inhibition activity of a non-RGD disintegrin from Crotalus durissus collilineatus venom

“…As to the primary sequence obtained through Edman degradation and mass spectrometry procedures, Cdc non-RGD disintegrin isolated in this study is absent from the classical RGD motif or any other motif previously observed in this protein family or glutamate-cysteine-aspartate (ECD) motif which characterize disintegrin-like proteins that may inhibit tumor progression [ 47 ]. In the same position of this domain, this toxin presents a triad of amino acids LVN, which was found in disintegrins of C. d. collilineatus by transcriptome techniques [ 48 ], as well as in the disintegrin domains of P-II SVMP of C. d. collilineatus (C0L2T8) and C. atrox (Q2QA03).…”

Breast Cancer Tumor Stroma: Cellular Components, Phenotypic Heterogeneity, Intercellular Communication, Prognostic Implications and Therapeutic Opportunities

“…In addition, integrins, such as αVβ3 cooperate with MMPs, such as MMP-2 [191], MMP-9 [192] and MMP-14 [193], in regulating migration of metastatic breast cancer cells toward specific substrates in an activation-dependent pathway. Indeed, it has been demonstrated that integrins control MMP-2 and MMP-9 expression regulating angiogenesis in breast tumor cells and endothelial cells [194]. In fact, the inhibition of MMP-9 and α V β 5 -integrin interaction results in a reduced angiogenesis and tumor invasion [195].…”

An update on angiogenesis modulators with a potential therapeutic role

“…The authors suggest that MDM2 might promote MMP9 driven angiogenesis [41]. Other experiments have indicated that the level of MMPs is modulated by α2β1 integrin in breast cancer and endothelial cells [42]. Webb et al (2017) have indicated that MMP9 and MMP2 promote the process of vascular metastasis and tumour growth in retinoblastoma, an ocular neoplasm described in children.…”