Glucose catabolism in cancer cells: identification and characterization of a marked activation response of the type II hexokinase gene to hypoxic conditions

J Biol Chem. 2001 Nov 16;276(46):43407-12. doi: 10.1074/jbc.M108181200. Epub 2001 Sep 13.


One of the most common signatures of highly malignant tumors is their capacity to metabolize more glucose to lactic acid than their tissues of origin. Hepatomas exhibiting this phenotype are dependent on the high expression of type II hexokinase, which supplies such tumors with abundant amounts of glucose 6-phosphate, a significant carbon and energy source especially under hypoxic conditions. Here we report that the distal region of the hepatoma type II hexokinase promoter displays consensus motifs for hypoxia-inducible factor (HIF-1) that overlap E-box sequences known to be related in other gene promoters to glucose response. Moreover, we show that subjecting transfected hepatoma cells to hypoxic conditions activates the type II hexokinase promoter almost 3-fold, a value that approaches 7-fold in the presence of glucose. Consistent with these findings is the induction under hypoxic conditions of the HIF-1 protein. Reporter gene analyses with a series of nested deletion mutants of the hepatoma type II hexokinase promoter show that a significant fraction of the total activation observed under hypoxic conditions localizes to the distal region where the overlapping HIF-1/E-box sequences are located. Finally, DNase I footprint analysis with a segment of the promoter containing these elements reveals the binding of several nuclear proteins. In summary, these novel studies identify and characterize a marked glucose-modulated activation response of the type II hexokinase gene to hypoxic conditions within highly glycolytic hepatoma cells, a property that may help assure that such cells exhibit a growth and survival advantage over their parental cells of origin.

Publication types

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

MeSH terms

  • Amino Acid Motifs
  • Animals
  • Base Sequence
  • Blotting, Western
  • Carcinoma, Hepatocellular / metabolism*
  • Cell Nucleus / metabolism
  • Cell Survival
  • Conserved Sequence
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Deoxyribonuclease I / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Activation
  • Exons
  • Female
  • Gene Deletion
  • Genes, Reporter
  • Glucose / metabolism*
  • Glucose-6-Phosphate / metabolism
  • Hexokinase / genetics*
  • Humans
  • Hypoxia*
  • Hypoxia-Inducible Factor 1
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Luciferases / metabolism
  • Models, Genetic
  • Molecular Sequence Data
  • Neoplasm Transplantation
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Oxygen / metabolism
  • Phenotype
  • Plasmids / metabolism
  • Promoter Regions, Genetic
  • Protein Binding
  • Rats
  • Transcription Factors*
  • Transfection
  • Tumor Cells, Cultured
  • beta-Galactosidase / metabolism


  • DNA-Binding Proteins
  • HIF1A protein, human
  • Hif1a protein, rat
  • Hypoxia-Inducible Factor 1
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Nuclear Proteins
  • Transcription Factors
  • Glucose-6-Phosphate
  • Luciferases
  • Hexokinase
  • Deoxyribonuclease I
  • beta-Galactosidase
  • Glucose
  • Oxygen

Associated data

  • GENBANK/U19605