Arresten, a Collagen-Derived Angiogenesis Inhibitor, Suppresses Invasion of Squamous Cell Carcinoma

Abstract

The turnover of extracellular matrix liberates various cryptic molecules with novel biological activity. Among these are the collagen-derived anti-angiogenic fragments, some of which are suggested to affect carcinoma cells also directly. Arresten is an endogenous angiogenesis inhibitor that is derived from the non-collagenous domain of the basement membrane collagen IV α1 chain. As the mere prevention of tumor angiogenesis leads to hypoxia that can result in selection of more aggressive cell types and reduces the efficacy of chemotherapy, we aimed here to elucidate how arresten influences the aggressive human carcinoma cells. Arresten efficiently inhibited migration and invasion of HSC-3 tongue carcinoma cells in culture and in an organotypic model. Subcutaneous Arr-HSC xenografts grew markedly more slowly in nude mice and showed reduced tumor cell proliferation, vessel density and local invasiveness. In the organotypic assay, HSC-3 cells overproducing arresten (Arr-HSC) showed induction of cell death. In monolayer culture the Arr-HSC cells grew in aggregated cobblestone-like clusters and, relative to the control cells, showed increased expression and localization of epithelial marker E-cadherin in cell-cell contacts. Application of electric cell-substrate impedance sensing (ECIS) further supported our observations on altered morphology and motility of the Arr-HSC cells. Administration of a function-blocking α1 integrin antibody abolished the impedance difference between the Arr-HSC and control cells suggesting that the effect of arresten on promotion of HSC-3 cell-cell contacts and cell spreading is at least partly mediated by α1β1 integrin. Collectively, our data suggest novel roles for arresten in the regulation of oral squamous carcinoma cell proliferation, survival, motility and invasion through the modulation of cell differentiation state and integrin signaling.

The APC tumor suppressor is required for epithelial cell polarization and three-dimensional morphogenesis

“…Interestingly, down-regulation of Lipocalin2 (LCN2), a marker for kidney injury (Viau et al, 2010), implicated in kidney epithelial cell morphogenesis (Gwira et al, 2005) and previously shown to be dysregulated in Apc Min/+ intestinal adenomas (Reichling et al, 2005), was observed in the APC shRNA MDCK cells. Other dysregulated genes including the alpha 1 subunit of type IV collagen (COL4A1, a basement membrane component), C-X-C chemokine receptor type 7 (CXCR7), and ADAM metallopeptidase with thrombospondin type 1, motif 6 (ADAMTS6), are implicated in controlling cell-cell or cell-matrix interactions (Kuhn, 1995; Bevitt et al, 2005; Hou et al, 2010; Aikio et al, 2012). These data collectively support a model in which APC loss-of-function in epithelial cells (through mutation and deletion of its c-terminus or gene silencing) leads to loss of polarity and tissue architecture, and subsequent tumor initiation, via altered communication between neighboring cells and the substratum.…”

Tumour–stroma crosstalk in the development of squamous cell carcinoma

“…The non-collagenous domain of type IV collagen α1 also contains a 26-kDa fragment known as arresten that, once cleaved becomes an endogenous angiogenesis inhibitor shown to also efficiently inhibit proliferation, migration and invasion of a highly metastatic human tongue SCC cell line (Aikio, Alahuhta et al 2012). Arresten is also known to inhibit the proliferation, migration and tube formation of different types of endothelial cells through α1β1 integrin receptors (Nyberg, Xie et al 2008).…”

Investigation of Cell Migration and Invasion Using Real-time Cell Analysis, as well as the Association with Matrix Metalloproteinase-9 in Oral Squamous Cell Carcinomas

“…The binding affinity state of many integrins can be regulated at the cell surface by several stimuli, allowing regulation of the intensity of adhesion during cellular interactions and migration 39 . Several reports have shown that integrin 1 1 in particular is associated with migration and invasion of cancer cells 40,41 . In this study, SCC-4 cells in which integrin 1 mRNA was strongly expressed showed high migration and invasion abilities.…”