Suppression of matrix metalloproteinase-9 transcription by transforming growth factor-beta is mediated by a nuclear factor-kappaB site

Biochem J. 2004 Jul 15;381(Pt 2):413-22. doi: 10.1042/BJ20040058.


TGF-beta (transforming growth factor-beta) plays a critical role in modulating the inflammatory response and other biological processes through its regulation of the production of MMPs (matrix metalloproteinases). In both Mono-Mac-6 and RAW264.7 monocyte/macrophage cells, TGF-beta abrogated lipopolysaccharide-induced increases in the enzymic activity and mRNA level of MMP-9. A fragment of the human MMP-9 promoter was used to characterize its regulation by TGF-beta signalling. In RAW264.7 cells, TGF-beta or its downstream signalling protein, Smad3 (Sma- and Mad-related protein 3), inhibited lipopolysaccharide-stimulated promoter activity. The suppressive activity of TGF-beta on the MMP-9 promoter was abrogated by an inhibitory Smad, Smad7. The MMP-9 promoter contains a putative TIE (TGF-beta inhibitory element). However, neither mutation nor deletion of the TIE had any effect on the inhibitory activity of TGF-beta on MMP-9 transcription, indicating that the consensus TIE is not required for this effect of TGF-beta. Analysis using a series of deletion mutants of the MMP-9 promoter revealed that a region containing a consensus NF-kappaB (nuclear factor-kappaB) site is required for the basal activity and TGF-beta-mediated suppression of the promoter. Mutation of the putative NF-kappaB site not only markedly reduced the basal transcriptional activity of the promoter, but also abrogated the responsiveness of the promoter to TGF-beta. In addition, a minimal promoter containing one copy of the NF-kappaB sequence was responsive to TGF-beta treatment. Furthermore, an electrophoretic mobility shift assay was performed with the nuclear extracts from RAW264.7 cells, and it was found that TGF-beta treatment did not disrupt the binding of NF-kappaB p50 and p65 proteins to the NF-kappaB sequence. Taken together, these studies indicate that the NF-kappaB site is indispensable for the suppressive activity of TGF-beta in the regulation of MMP-9 transcription.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Binding Sites
  • Cell Line
  • Cell Line, Tumor
  • Gene Expression Regulation / physiology*
  • Humans
  • Leukemia, Monocytic, Acute / genetics
  • Leukemia, Monocytic, Acute / metabolism
  • Leukemia, Monocytic, Acute / pathology
  • Macrophages / chemistry
  • Macrophages / metabolism
  • Matrix Metalloproteinase 9 / genetics*
  • Matrix Metalloproteinase 9 / metabolism
  • Monocytes / chemistry
  • Monocytes / metabolism
  • NF-kappa B / metabolism
  • NF-kappa B / physiology*
  • NF-kappa B p50 Subunit
  • Promoter Regions, Genetic / physiology
  • Protein Binding
  • Signal Transduction / physiology
  • Transcription Factor RelA
  • Transcription, Genetic / physiology
  • Transforming Growth Factor beta / metabolism
  • Transforming Growth Factor beta / physiology*


  • NF-kappa B
  • NF-kappa B p50 Subunit
  • Transcription Factor RelA
  • Transforming Growth Factor beta
  • Matrix Metalloproteinase 9