Lamininα3 LG4 Module Induces Keratinocyte Migration: Involvement of Matrix Metalloproteinase-9

Abstract

Laminin alpha3 chain, a functionally key subunit of laminin-5, contains a large globular module (G module) which consists of a tandem repeat of five homologous LG modules (LG1-5). We previously demonstrated that the LG4 module of laminin alpha3 chain (alpha3 LG4) induces a matrix metalloproteinase-1 (MMP-1) expression through the interaction with syndecans leading to MAPK activation/IL-1beta expression signaling loop (Utani et al., J. Biol. Chem. 278, 34483-34490, 2003). Here, we show that a recombinant alpha3 LG4 and synthetic peptides containing syndecan binding motif induced a cell motility and a MMP-9 expression in ketarinocytes. The synthetic peptide (A3G756)-induced cell migration and MMP-9 upregulation were inhibited by each application of a heparin and an IL-1 receptor antagonist (IL-1RA), suggesting the involvement of syndecans and IL-1beta autocrine. Furthermore, the A3G756-induced cell motility was inhibited by an MMP-9 inhibitor and a neutralizing antibody of MMP-9, indicating induced cell motility was dependent on an MMP-9 activity. Taken these together, laminin-5 alpha3 LG4 module may play an important role in re-epithelialization at tissue remodeling.

Targeting a Tumor-Specific Laminin Domain Critical for Human Carcinogenesis

“…Our observation extends earlier findings of the role of G45 domain in up-regulating MMP-1 (46) and MMP-9 in keratinocytes to squamous carcinoma cells (47). These results are consistent with other studies showing the role of extracellular matrix molecules changing MMP expression and activities (48).…”

Reduced migration, altered matrix and enhanced TGFβ1 signaling are signatures of mouse keratinocytes lacking Sdc1

“…(Hayashida et al, 2006). In response to injury in vivo, genes for TGF␤1 and Sdc1 are upregulated in both epithelial and mesenchymal cells, and the Sdc1 ectodomain is shed at wound sites where the soluble molecule can regulate chemokine function (Gotte and Echtermeyer, 2003;Tkachenko et al, 2004) and modulate the activation of various MMPs (Kelly et al, 2000;Steffensen et al, 2001;Momota et al, 2005). The role Sdc1 plays in cancer is complex and tissue specific, but recent studies using Sdc1-null mice show that these mice are resistant to mammary carcinogenesis (Alexander et al, 2002;McDermott et al, 2007).…”

Cell Surface Proteoglycans Syndecan-1 and -4 Bind Overlapping but Distinct Sites in Laminin α3 LG45 Protein Domain

“…We previously showed that syndecan-4 is not involved in this mechanism (30), and we confirmed here that residues specific for the syndecan-4 interaction (Lys-1440/ Lys-1442) were not involved in syndecan-1-mediated cell adhesion. Other in vitro studies reported that adding either recombinant ␣3LG45 or LG4 to the culture medium could induce migration of NHK (26). It was further suggested that this migration depended on a specific sequence within LG4 ( 1412 NSFMALYL-SKGR; see Ref.…”

“…LG45 on Keratinocyte Migration-A study reported that LG45 could stimulate NHK migration when added to the culture through a mechanism involving syndecan-4 (26,27). Therefore, we tested whether mutation in the syndecan-4-binding site could affect this phenomenon.…”

“…These HBDs conferred heparin-dependent cell adhesion properties, which suggested that this region in LN332 could interact with an HSPG cellular receptor (20 -22). Later, a motif in the LG45 domain that included residues 1412 NSFMALYLSKGR was shown to induce syndecan-2-and -4-mediated cell adhesion, neurite outgrowth, and matrix metalloproteinase-1 and -9 secretion (23)(24)(25)(26). Further work suggested that this peptide motif also induced keratinocyte migration by triggering syndecan-4 clustering and subsequent ␤1 integrin activation (27).…”