HGF potentiates extracellular matrix-driven migration of human myoblasts: involvement of matrix metalloproteinases and MAPK/ERK pathway

Abstract

BackgroundThe hepatocyte growth factor (HGF) is required for the activation of muscle progenitor cells called satellite cells (SC), plays a role in the migration of proliferating SC (myoblasts), and is present as a soluble factor during muscle regeneration, along with extracellular matrix (ECM) molecules. In this study, we aimed at determining whether HGF is able to interact with ECM proteins, particularly laminin 111 and fibronectin, and to modulate human myoblast migration.MethodsWe evaluated the expression of the HGF-receptor c-Met, laminin, and fibronectin receptors by immunoblotting, flow cytometry, or immunofluorescence and used Transwell assays to analyze myoblast migration on laminin 111 and fibronectin in the absence or presence of HGF. Zymography was used to check whether HGF could modulate the production of matrix metalloproteinases by human myoblasts, and the activation of MAPK/ERK pathways was evaluated by immunoblotting.ResultsWe demonstrated that human myoblasts express c-Met, together with laminin and fibronectin receptors. We observed that human laminin 111 and fibronectin have a chemotactic effect on myoblast migration, and this was synergistically increased when low doses of HGF were added. We detected an increase in MMP-2 activity in myoblasts treated with HGF. Conversely, MMP-2 inhibition decreased the HGF-associated stimulation of cell migration triggered by laminin or fibronectin. HGF treatment also induced in human myoblasts activation of MAPK/ERK pathways, whose specific inhibition decreased the HGF-associated stimulus of cell migration triggered by laminin 111 or fibronectin.ConclusionsWe demonstrate that HGF induces ERK phosphorylation and MMP production, thus stimulating human myoblast migration on ECM molecules. Conceptually, these data state that the mechanisms involved in the migration of human myoblasts comprise both soluble and insoluble moieties. This should be taken into account to optimize the design of therapeutic cell transplantation strategies by improving the migration of donor cells within the host tissue, a main issue regarding this approach.Electronic supplementary materialThe online version of this article (10.1186/s13395-017-0138-6) contains supplementary material, which is available to authorized users.

Extracellular Matrix Influencing HGF/c-MET Signaling Pathway: Impact on Cancer Progression

“…scatter factor) is a pleiotropic cytokine initially synthesized and secreted as a biologically inactive single-chain precursor (pro-HGF) [ 36 , 37 ]. However, when extracellular proteases (e.g., serine proteases and MMPs) cleave the Arg494-Val495 peptide bond of pro-HGF, this biologically inactive precursor is converted into its bioactive form [ 38 , 39 , 40 ]. Mature HGF is a disulfide-linked heterodimer composed of an α- and β-chain.…”

Formin-like 3 regulates RhoC/FAK pathway and actin assembly to promote cell invasion in colorectal carcinoma

“…The levels of MMP-2 and MMP-9 were regulated by phosphorylated MAPK and AKT[ 29 , 30 ], which were activated by RhoC[ 31 - 33 ] or in sequence activated by FAK, Pyk2 and RhoC[ 25 ]. Moreover, the expression of VEGF and MMP-9 were inhibited by the down-regulation of RhoC[ 28 ].…”

“…Both MMP-2 and MMP-9 were shown to be activated by phosphorylated MAPK and AKT[ 29 , 30 ]. In addition, MAPK and AKT signaling pathways were activated by RhoC[ 31 - 33 ] or in sequence by FAK, Pyk2 and RhoC[ 25 ].…”

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