Fibroblast-directed expression and localization of 92-kDa type IV collagenase along the tumor-stroma interface in an in vitro three-dimensional model of human squamous cell carcinoma

Abstract

The malignant dissemination of tumors has been shown to require expression of one or more members of the matrix metalloprotease (MMP) enzyme family, whose function is to catalyze degradation of extracellular matrix proteins. In human squamous cell carcinoma (SCC) of the skin, expression of the MMP 92-kDa type IV collagenase (MMP-9), was previously shown to localize to malignant keratinocytes residing along the tumor/stromal interface. The purpose of the study presented here was to determine whether this localized expression pattern is due to interactions between SCC cells and adjacent stromal fibroblasts. To examine this question, SCC cells were grown as organotypic skin cultures, an in vitro three-dimensional model of reconstructed human epidermis in which keratinocytes are grown on a type 1 collagen gel embedded with human dermal fibroblasts. In this study, MMP-9 expression was compared in organotypic cultures (constructed with SCC cells or the non-tumorigenic keratinocyte cell line HaCaT), in which human dermal fibroblasts were either included or excluded from the underlying stromal layer. In the absence of fibroblasts, expression of MMP-9 was slightly higher in SCC than HaCaT cultures. In cultures constructed with fibroblasts, however, induction of MMP-9 mRNA was observed in SCC but not HaCaT cultures. This induction of MMP-9 mRNA was accompanied by high levels of MMP-9 protein expression along the SCC/stromal interface. These data provide strong evidence that interactions between malignant keratinocytes and adjacent stromal fibroblasts are critical in directing expression of MMP-9 to the tumor-stroma interface in human SCC tumors.