title: Expression of matrix metalloproteinase-2 and -9 and membrane-type 1 matrix metalloproteinase in melanocytic tumors of dogs and canine melanoma cell lines.
abstract: OBJECTIVE
  To evaluate expression of matrix metalloproteinase (MMP)-2 and -9 and membrane-type 1 MMP (MT1-MMP) in melanocytomas and malignant melanomas of dogs, analyze in vitro production of MMPs by canine melanoma cell lines and primary dermal fibroblasts, and investigate mutual communication between tumor cells and fibroblasts and the influence of collagen on MMP regulation.
  
  
  METHODS
  35 biopsy specimens from melanocytic tumors and primary dermal fibroblasts of dogs and 3 canine melanoma cell lines (CML-1, CML-10c2, and CML-6M).
  
  
  METHODS
  MMP-2, MMP-9, and MT1-MMP were detected in tumor samples by use of immunohistochemical analysis. In vitro production was analyzed via reverse transcriptase-PCR assay, immunocytochemical analysis, zymography, and immunoblotting.
  
  
  RESULTS
  MMP-9 was overexpressed in malignant melanomas, compared with expression in melanocytomas, whereas no significant differences in MMP-2 and MT1-MMP immunostaining were detected. Stromal cells also often had positive staining results. In vitro, all 3 melanoma cell lines and dermal fibroblasts had evidence of MMP-2 and MT1-MMP, but only melanoma cells had evidence of MMP-9. Coculture of CML-1 or CML-10c2 cells and dermal fibroblasts induced an increase in expression of the active form of MMP-2. Culture of melanoma cells on type I collagen increased the activation state of MT1-MMP.
  
  
  CONCLUSIONS
  MMP-9 expression was increased in malignant melanomas of dogs. Stromal cells were a source for MMPs. Stromal cells, in combination with matrix components such as type I collagen, can interact with tumor cells to regulate MMP production. Information about MMP production and regulation could help in the development of new treatments.
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Facultat de Veterinària, Universitat Autònoma de Barcelona, 08193 Cerdanyola del Vallès, Barcelona, Spain.
authors.forename: María-José
authors.initials: MJ
authors.lastname: Docampo
authors.forename: Jennifer
authors.initials: J
authors.lastname: Cabrera
authors.forename: Rosa M
authors.initials: RM
authors.lastname: Rabanal
authors.forename: Anna
authors.initials: A
authors.lastname: Bassols
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D004283:Dog Diseases / Q000201:enzymology* / Q000473:pathology; D004285:Dogs; D005347:Fibroblasts / Q000201:enzymology / Q000473:pathology; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D018960:Hyaluronan Receptors / Q000032:analysis / Q000378:metabolism; D006820:Hyaluronic Acid / Q000032:analysis / Q000378:metabolism; D007124:Immunoenzyme Techniques / Q000662:veterinary; D053511:Matrix Metalloproteinase 14 / Q000032:analysis / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000032:analysis / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000032:analysis / Q000378:metabolism*; D020782:Matrix Metalloproteinases / Q000032:analysis / Q000378:metabolism*; D008545:Melanoma / Q000201:enzymology / Q000473:pathology / Q000662:veterinary*; D012867:Skin / Q000201:enzymology / Q000473:pathology; D012878:Skin Neoplasms / Q000201:enzymology / Q000473:pathology / Q000662:veterinary*
chemical_list: D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid; D020782:Matrix Metalloproteinases; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D053511:Matrix Metalloproteinase 14