HER-2/neu represses the metastasis suppressor RECK via ERK and Sp transcription factors to promote cell invasion

J Biol Chem. 2006 Feb 24;281(8):4718-25. doi: 10.1074/jbc.M510937200. Epub 2005 Dec 23.

Abstract

Matrix metalloproteinase (MMP) inhibitory proteins may negatively regulate MMP activity to suppress tumor metastasis. In this study, we demonstrate that the HER-2/neu oncogene inhibits the expression of the MMP inhibitor RECK to promote cell invasion. RECK was inhibited via transcriptional repression in B104-1-1 cells, which express constitutively active HER-2/neu. Overexpression of HER-2/neu in NIH/3T3 or HaCaT cells also suppressed RECK expression. Deletion and mutation assays showed that HER-2/neu repressed RECK via the Sp1-binding site localized in the -82/-71 region from the translational start site. DNA affinity precipitation and chromatin immunoprecipitation assays indicated that binding of Sp1 and Sp3 to this consensus site was increased in B104-1-1 cells. We also found that HER-2/neu inhibited RECK via the ERK signaling pathway. Sp1 proteins phosphorylated at Thr453 and Thr739 by ERK bound preferentially to the RECK promoter, and this binding was reversed by HER-2/neu and ERK inhibitors. Furthermore, our data indicate that HER-2/neu obviously increased HDAC1 binding to the Sp1-binding site localized in the -82/-71 region of the RECK promoter. The histone deacetylase inhibitor trichostatin A reversed HER-2/neu-induced inhibition of RECK. HER-2/neu activation was associated with increased MMP-9 secretion and activation. Re-expression of RECK in HER-2/neu-overexpressing cells inhibited MMP-9 secretion and cell invasion. Taken together, our results suggest that HER-2/neu induces the binding of Sp proteins and HDAC1 to the RECK promoter to inhibit RECK expression and to promote cell invasion. Restoration of RECK provides a novel strategy for the inhibition of HER-2/neu-induced metastasis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Blotting, Western
  • Cell Line, Tumor
  • Cell Nucleus / metabolism
  • Chromatin Immunoprecipitation
  • DNA / metabolism
  • DNA, Complementary / metabolism
  • Down-Regulation
  • Extracellular Signal-Regulated MAP Kinases / metabolism*
  • GPI-Linked Proteins
  • Gene Deletion
  • Gene Expression Regulation, Neoplastic*
  • Genetic Vectors
  • Histone Deacetylase Inhibitors
  • Humans
  • Hydroxamic Acids / pharmacology
  • Immunoprecipitation
  • Matrix Metalloproteinase 9 / metabolism
  • Membrane Glycoproteins / metabolism*
  • Mice
  • NIH 3T3 Cells
  • Neoplasm Invasiveness
  • Neoplasm Metastasis
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Biosynthesis
  • Receptor, ErbB-2 / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Sp1 Transcription Factor / metabolism*
  • Sp3 Transcription Factor / metabolism
  • Threonine / chemistry
  • Transcription, Genetic
  • Transfection
  • Up-Regulation

Substances

  • DNA, Complementary
  • GPI-Linked Proteins
  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • Membrane Glycoproteins
  • RECK protein, human
  • Reck protein, mouse
  • Sp1 Transcription Factor
  • Sp3 Transcription Factor
  • Threonine
  • trichostatin A
  • DNA
  • Receptor, ErbB-2
  • Extracellular Signal-Regulated MAP Kinases
  • Matrix Metalloproteinase 9