Comparative studies on the secretion and activation of MMPs in two reconstructed human skin models using HaCaT- and HaCaT-ras-transfected cell lines
| Autor: | Claude Vincent, Mireille Serres, D. Schmitt, Marie-Jeanne Staquet, D. Nova, C. Le Griel, E. Gentilhomme, Sébastien Holvöet |
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| Rok vydání: | 2004 |
| Předmět: |
Keratinocytes
Cancer Research Cell Culture Techniques Human skin Matrix metalloproteinase Models Biological Cell Line Dermis medicine Tumor Cells Cultured Skin equivalent Humans Cell Line Transformed Skin integumentary system Chemistry General Medicine Fibroblasts Matrix Metalloproteinases Cell biology Enzyme Activation HaCaT medicine.anatomical_structure Genes ras Oncology Cell culture Immunology Collagenase Collagen Wound healing medicine.drug |
| Zdroj: | Clinicalexperimental metastasis. 20(8) |
| ISSN: | 0262-0898 |
| Popis: | Matrix metalloproteinases play an important role in tissue regeneration, wound healing and tumor invasion. Our previous studies have shown a higher motility of HaCaT-ras-transfected cells compared with HaCaT or normal human keratinocytes (NHK) in correlation with a higher secretion of MMP-2 (72 kDa) or MMP-9 (92 kDa), according to the medium used for cell cultures. Presently, the expression and activity of MMPs were investigated in two reconstructed skin models, using a dead de-epidermized dermis (DED) or a dermal substitute including living fibroblasts. In all experiments, MMP-9 was essentially secreted by NHK and to a greater extent by HaCaT cells. Its active form (86 kDa) was only detected in both reconstructed skin models according to keratinocyte differentiation. MMP-2 was mainly secreted by living fibroblasts included in the dermal substitute skin model. In this case, its activation was up-regulated when HaCaT cell lines were seeded onto the dermal substitute according to their culture at air/liquid interface as shown for MMP-9. The collagenase MMP-1 and stromelysin-1 (MMP-3), susceptible to activate pro-MMP-2 and -9, respectively, were detected in their inactive form by ELISA. MMP-1 was expressed in both models but MMP-3 required the presence of living fibroblasts. Their activities were not detected using specific fluorogenic substrates. In the skin equivalent model using HaCaT, the extensive secretion and activation of MMP-2 and MMP-9 could explain the defect observed in basal membrane reconstruction, suggesting a direct interaction of HaCaT with fibroblasts. |
| Databáze: | OpenAIRE |
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