Cloning and characterisation of a 1.1 kb fragment of the carcinoma-associated epithelial cell adhesion molecule promoter

Anticancer Res. 2003 Jul-Aug;23(4):3255-61.

Abstract

The epithelial cell adhesion molecule (EpCAM) is a transmembrane protein associated with a variety of carcinomas, where EpCAM is often strongly up-regulated or, as in the case of squamous cell carcinomas, de novo expressed. The molecular mechanisms underlying the transcriptional regulation of EpCAM are poorly understood. So far, a 570 bp fragment has been cloned and shown to have specific transcriptional activity, which was negatively-regulated upon the induction of the transcription factor NF-kappa B. In the present study we have cloned a 1100 bp fragment of the EpCAM promoter containing the 570 bp fragment and additional 550 bp upstream. We demonstrate that both fragments have strong synergistic effects with respect to transcriptional activity in EpCAM-positive cells. Furthermore, the 1100 bp fragment was likewise negatively-regulated upon TNF alpha and IFN alpha treatment, thus retaining silencer sequences.

MeSH terms

  • Antigens, Neoplasm / genetics*
  • Carcinoma / genetics*
  • Carcinoma / metabolism
  • Cell Adhesion Molecules / genetics*
  • Cloning, Molecular
  • Epithelial Cell Adhesion Molecule
  • Flow Cytometry
  • Humans
  • Interferon-alpha / pharmacology
  • Interleukin-4 / pharmacology
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic / drug effects
  • Transcriptional Activation / drug effects
  • Transfection
  • Tumor Cells, Cultured
  • Tumor Necrosis Factor-alpha / pharmacology

Substances

  • Antigens, Neoplasm
  • Cell Adhesion Molecules
  • Epithelial Cell Adhesion Molecule
  • Interferon-alpha
  • Tumor Necrosis Factor-alpha
  • Interleukin-4