Abundant expression of Dec1/stra13/sharp2 in colon carcinoma: its antagonizing role in serum deprivation-induced apoptosis and selective inhibition of procaspase activation

Biochem J. 2002 Oct 15;367(Pt 2):413-22. doi: 10.1042/BJ20020514.

Abstract

The basic helix-loop-helix (bHLH) proteins are intimately associated with developmental events such as cell differentiation and lineage commitment. The HLH domain in the bHLH motif is responsible for dimerization, whereas the basic region mediates DNA binding. Based on sequence alignment and domain analysis, differentially expressed in chondrocytes/stimulated with retinoic acid/split and hairy-related proteins (DEC/STRA/SHARPs) represent a new class of bHLH proteins. The present study describes the functional characterization of DEC1. Subtractive experiments and blotting analyses demonstrated that DEC1 was highly expressed in colon carcinomas, but not in the adjacent normal tissues. Several cell cycle blockers markedly induced DEC1 expression. Stable transfectants with a tetracycline-inducible construct demonstrated that DEC1 caused proliferation inhibition, antagonized serum deprivation-induced apoptosis and selectively inhibited the activation of procaspases. These activities were highly correlated with the abundance of tetracycline-induced DEC1. Stable transfectants expressing a mutant DEC1 (lacking the DNA-binding domain) exhibited neither proliferation inhibition nor apoptotic antagonism, which suggests that DNA binding is required for these actions. Enzymic assays and immunoblotting analyses demonstrated that induction of DEC1 by tetracycline significantly decreased the activation of procaspases 3, 7 and 9 but not procaspase 8. The selective suppression on the activation of procaspases 3, 7 and 9 over procaspase 8 suggests that DEC1-mediated anti-apoptosis is achieved by blocking apoptotic pathways initiated via the mitochondria. The results functionally distinguish DEC1 from other bHLH proteins and directly link this factor to oncogenesis.

Publication types

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

MeSH terms

  • Adenocarcinoma / genetics*
  • Adenocarcinoma / pathology
  • Adult
  • Amino Acid Sequence
  • Apoptosis / drug effects
  • Apoptosis / physiology
  • Basic Helix-Loop-Helix Transcription Factors
  • Caspase 3
  • Caspase 7
  • Caspase 8
  • Caspase 9
  • Caspases / drug effects
  • Caspases / genetics
  • Caspases / metabolism*
  • Cell Cycle / drug effects
  • Cell Division / genetics
  • Colonic Neoplasms / genetics*
  • Colonic Neoplasms / pathology
  • Culture Media, Serum-Free / pharmacology
  • Enzyme Activation / drug effects
  • Enzyme Precursors / drug effects
  • Enzyme Precursors / genetics
  • Enzyme Precursors / metabolism*
  • Female
  • Gene Expression Regulation, Neoplastic
  • Helix-Loop-Helix Motifs
  • Homeodomain Proteins / genetics*
  • Homeodomain Proteins / metabolism
  • Humans
  • Hydroxyurea / pharmacology
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Mutation

Substances

  • BHLHE40 protein, human
  • Basic Helix-Loop-Helix Transcription Factors
  • Culture Media, Serum-Free
  • Enzyme Precursors
  • Homeodomain Proteins
  • CASP3 protein, human
  • CASP7 protein, human
  • CASP8 protein, human
  • CASP9 protein, human
  • Caspase 3
  • Caspase 7
  • Caspase 8
  • Caspase 9
  • Caspases
  • Hydroxyurea