title: TGF-beta1 + EGF-initiated invasive potential in transformed human keratinocytes is coupled to a plasmin/MMP-10/MMP-1-dependent collagen remodeling axis: role for PAI-1.
abstract: The phenotypic switching called epithelial-to-mesenchymal transition is frequently associated with epithelial tumor cell progression from a comparatively benign to an aggressive, invasive malignancy. Coincident with the emergence of such cellular plasticity is an altered response to transforming growth factor-beta (TGF-beta) as well as epidermal growth factor (EGF) receptor amplification. TGF-beta in the tumor microenvironment promotes invasive traits largely through reprogramming gene expression, which paradoxically supports matrix-disruptive as well as stabilizing processes. ras-transformed HaCaT II-4 keratinocytes undergo phenotypic changes typical of epithelial-to-mesenchymal transition, acquire a collagenolytic phenotype, and effectively invade collagen type 1 gels as a consequence of TGF-beta1 + EGF stimulation in a three-dimensional physiologically relevant model system that monitors collagen remodeling. Enhanced collagen degradation was coupled to a significant increase in matrix metalloproteinase (MMP)-10 expression and involved a proteolytic axis composed of plasmin, MMP-10, and MMP-1. Neutralization of any one component in this cascade inhibited collagen gel lysis. Similarly, addition of plasminogen activator inhibitor type 1 (SERPINE1) blocked collagen degradation as well as the conversion of both proMMP-10 and proMMP-1 to their catalytically active forms. This study therefore identifies an important mechanism in TGF-beta1 + EGF-initiated collagen remodeling by transformed human keratinocytes and proposes a crucial upstream role for plasminogen activator inhibitor type 1-dependent regulation in this event.
authors.affiliation: Center for Cell Biology and Cancer Research, Albany Medical College, Albany, New York 12208, USA.
authors.forename: Cynthia E
authors.initials: CE
authors.lastname: Wilkins-Port
authors.forename: Qunhui
authors.initials: Q
authors.lastname: Ye
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Mazurkiewicz
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Higgins
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology; D002461:Cell Line, Transformed; D024042:Collagen Type I / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005341:Fibrinolysin / Q000378:metabolism*; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism*; D053506:Matrix Metalloproteinase 10 / Q000378:metabolism*; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism*; D012878:Skin Neoplasms / Q000378:metabolism / Q000473:pathology; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*
chemical_list: D024042:Collagen Type I; D017395:Plasminogen Activator Inhibitor 1; C051321:SERPINE1 protein, human; D053773:Transforming Growth Factor beta1; D004815:Epidermal Growth Factor; D005341:Fibrinolysin; D053506:Matrix Metalloproteinase 10; D020781:Matrix Metalloproteinase 1