Interleukin 10 blocks matrix metalloproteinase-2 and membrane type 1-matrix metalloproteinase synthesis in primary human prostate tumor lines

Clin Cancer Res. 2003 Mar;9(3):1191-9.

Abstract

Insulin-like growth factor (IGF) I has been shown previously to up-regulate matrix metalloproteinase-2 (MMP-2) production, whereas the interleukin (IL) 10/IL-10 receptor axis has been found to down-regulate MMP-2 synthesis in tumor cells. In this paper, we showed that IL-10 activation of the IL-10 receptor blocked MMP-2 and membrane type 1 (MT1) -MMP transcription and protein synthesis in nonimmortalized primary human prostate cell strains (i.e., HPCA-10a and HPCA-10c) derived from high-grade cancer. Northern blots, Western blots, and ELISAs showed that IL-10 suppressed IGF-I induction of MMP-2 and MT1-MMP mRNA synthesis in these cell strains (P < 0.001). Inhibition studies with IL-10 and IGF-I receptor antibodies plus transfections experiments with IL-10 sense, and IGF-I receptor antisense constructs confirmed these results. Finally, transient transfection experiments and chloramphenicol acetyltransferase assays with different regions of the 5' promoter region of the MMP-2 gene (-1659 to -555 bp) additionally showed that IGF-I stimulated p53-dependent plasmid catecholamine acetyltransferase activity and that IL-10 blocked IGF-I-induced plasmid catecholamine acetyltransferase activity. Electrophoretic mobility shift assays revealed that IL-10 induced protein(s) binding to a putative "silencer element" (-1309 to -555 fragment) downstream of the p53 binding site (-1649 to -1640). The data show that IL-10 blocks IGF-I activation of MMP-2 and MT1-MMP mRNA expression and protein synthesis in primary prostate cell strains.

MeSH terms

  • Binding Sites
  • Blotting, Northern
  • Blotting, Western
  • Chloramphenicol O-Acetyltransferase / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Insulin-Like Growth Factor I / metabolism
  • Interleukin-10 / metabolism
  • Interleukin-10 / pharmacology*
  • Male
  • Matrix Metalloproteinase Inhibitors*
  • Matrix Metalloproteinases, Membrane-Associated
  • Metalloendopeptidases / antagonists & inhibitors*
  • Metalloendopeptidases / metabolism
  • Oligonucleotides, Antisense / pharmacology
  • Plasmids / metabolism
  • Promoter Regions, Genetic
  • Prostatic Neoplasms
  • RNA, Messenger / metabolism
  • Receptors, Interleukin / metabolism
  • Receptors, Interleukin-10
  • Transcription, Genetic
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Matrix Metalloproteinase Inhibitors
  • Oligonucleotides, Antisense
  • RNA, Messenger
  • Receptors, Interleukin
  • Receptors, Interleukin-10
  • Interleukin-10
  • Insulin-Like Growth Factor I
  • Chloramphenicol O-Acetyltransferase
  • Matrix Metalloproteinases, Membrane-Associated
  • Metalloendopeptidases