Dominant-negative CREB inhibits heparanase functionality and melanoma cell invasion

J Cell Biochem. 2004 Oct 1;93(2):215-23. doi: 10.1002/jcb.20231.


Heparanase (HPSE-1) is an endo-beta-D-glucuronidase involved in the degradation of cell-surface/extracellular matrix heparan sulfate (HS) in normal and neoplastic tissues. HPSE-1 represents the first example of purification and cloning of a mammalian HS-degradative enzyme. Elevated HPSE-1 levels are known to be associated with metastatic cancers, directly implicating HPSE-1 in metastatic events. The purpose of this study was to determine the role of cAMP response element-binding protein (CREB) in modulating HPSE-1-mediated effects on human melanoma cell invasion. Highly invasive, brain-metastatic melanoma cells (70W) were transfected with the dominant-negative CREB (KCREB) and subsequently analyzed for changes in their HPSE-1 content, functionality, and cell invasive properties. KCREB-transfected cells showed a decrease in HPSE-1 mRNA expression and activity. This correlated with a significantly decreased invasion of these cells through Matrigel-coated filters. Furthermore, adenoviral vectors containing the full-length human HPSE-1 cDNA in sense orientation (Ad-S/hep) were constructed to investigate CREB effects on HPSE-1. Restoration of HPSE-1 expression and functionality following Ad-S/hep infection of KCREB-transfected 70W cells recovered melanoma cell invasiveness. These results demonstrate that KCREB inhibits HPSE-1 and suggest that one of the roles CREB plays in the acquisition of melanoma cells metastatic phenotype is affecting HPSE-1 activity.

Publication types

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

MeSH terms

  • Adenoviridae / genetics
  • Cell Line, Tumor
  • Collagen
  • Cyclic AMP / metabolism
  • Cyclic AMP Response Element-Binding Protein / genetics*
  • Cyclic AMP Response Element-Binding Protein / metabolism*
  • DNA, Complementary / genetics
  • Drug Combinations
  • Gene Expression Regulation, Neoplastic
  • Genes, Dominant / genetics
  • Glucuronidase / antagonists & inhibitors*
  • Glucuronidase / genetics
  • Glucuronidase / metabolism*
  • Humans
  • Laminin
  • Melanoma / enzymology
  • Melanoma / genetics
  • Melanoma / metabolism*
  • Melanoma / pathology*
  • Mutation / genetics
  • Neoplasm Invasiveness / genetics
  • Neoplasm Invasiveness / pathology
  • Proteoglycans
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Transfection


  • Cyclic AMP Response Element-Binding Protein
  • DNA, Complementary
  • Drug Combinations
  • Laminin
  • Proteoglycans
  • RNA, Messenger
  • matrigel
  • Collagen
  • Cyclic AMP
  • heparanase
  • Glucuronidase