Effect of phorbol ester and cytokines on matrix metalloproteinase and tissue inhibitor of metalloproteinase expression in tumor and normal cell lines

Invasion Metastasis. 1992;12(3-4):168-84.


The effect of the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) and the cytokines interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) on matrix metalloproteinases (MMP) and metalloproteinase inhibitors was studied in a variety of human cell lines. Expression of the mammalian collagenase (MMP-1), 72-kD gelatinase/type IV collagenase (MMP-2), stromelysin (MMP-3), 92-kD gelatinase/type IV collagenase (MMP-9), and tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) was assessed by zymography and Northern blot analysis. MMP-2 and TIMP-2 activities were refractory to TPA, IL-1 and TNF-alpha treatment in most of the cell lines. In contrast, MMP-3, MMP-9 and TIMP-1 activities were markedly stimulated by TPA in most of the tumor cell lines and human umbilical vein endothelial cells (HUVEC), whereas the fibroblast lines were minimally stimulated or unresponsive to TPA. The MMP-3, MMP-9 and TIMP-1 stimulation in response to IL-1 and TNF-alpha treatment was detected in some of the tumor cell lines and HUVEC. The increase in activity was less marked than in TPA. A breast carcinoma cell line, MDA-MB-231, which did not express MMP-2, had high expression of MMP-3 and MMP-9 which were unaffected by TPA and cytokine treatment. Northern blot analysis of MMP and TIMP mRNA expression reflected the zymogram findings for most of the cell lines. TPA-mediated stimulation of MMP-1 was similar to that of MMP-3 and MMP-9. Exceptions were the fibroblast cell lines which showed either a much more marked mRNA response of MMP-9 to TPA than observed at protein level, or a high constitutive MMP-9 mRNA when MMP-9 activity was not detectable by zymography. TPA-mediated stimulation of MMP-9 and TIMP-1 activity was blocked by staurosporine, an inhibitor of protein kinase C (PKC). A non-PKC-activating phorbol ester, 4 alpha-phorbol-12,13-didecanoate, did not stimulate MMP-9 and TIMP-1 activity. TPA treatment caused the increased expression of c-fos containing AP-1-specific binding activity in selected tumor cell lines. This activity was maximal at 6 h. An association was observed between AP-1 binding activity and increased expression of MMP-1, MMP-3 and MMP-9, which possess TPA-responsive elements (TRE). TPA-sensitive MMPs and TIMP-1 were variably stimulated by biologically relevant cytokines, such as IL-1 and TNF-alpha.(ABSTRACT TRUNCATED AT 400 WORDS)

Publication types

  • Comparative Study

MeSH terms

  • Base Sequence
  • Blotting, Northern
  • Collagenases / drug effects
  • Collagenases / metabolism
  • Cycloheximide / pharmacology
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / enzymology
  • Fibroblasts / drug effects
  • Fibroblasts / enzymology
  • Glycoproteins / drug effects*
  • Glycoproteins / metabolism
  • Humans
  • Interleukin-1 / pharmacology*
  • Matrix Metalloproteinase 1
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 3
  • Matrix Metalloproteinase 9
  • Metalloendopeptidases / drug effects*
  • Metalloendopeptidases / metabolism
  • Molecular Sequence Data
  • Neoplasm Proteins / drug effects*
  • Neoplasm Proteins / metabolism
  • Neoplasms / enzymology
  • Protein Kinase C / antagonists & inhibitors
  • Proto-Oncogene Proteins c-jun / metabolism
  • RNA / metabolism
  • Sensitivity and Specificity
  • Tetradecanoylphorbol Acetate / pharmacology*
  • Tissue Inhibitor of Metalloproteinase-2
  • Tissue Inhibitor of Metalloproteinases
  • Transcription, Genetic
  • Tumor Cells, Cultured / drug effects
  • Tumor Necrosis Factor-alpha / pharmacology*


  • Glycoproteins
  • Interleukin-1
  • Neoplasm Proteins
  • Proto-Oncogene Proteins c-jun
  • Tissue Inhibitor of Metalloproteinases
  • Tumor Necrosis Factor-alpha
  • Tissue Inhibitor of Metalloproteinase-2
  • RNA
  • Cycloheximide
  • Protein Kinase C
  • Collagenases
  • Metalloendopeptidases
  • Matrix Metalloproteinase 3
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9
  • Matrix Metalloproteinase 1
  • Tetradecanoylphorbol Acetate