Use of the EGP-2/Ep-CAM promoter for targeted expression of heterologous genes in carcinoma derived cell lines

Cancer Gene Ther. 2004 Sep;11(9):603-12. doi: 10.1038/sj.cgt.7700725.

Abstract

EGP-2, also known as Ep-CAM, is expressed at high levels on the surface of most carcinomas and is therefore considered an attractive target for anticancer strategies. To explore the mechanisms regulating the expression of EGP-2, sequences 3.4 kb upstream of the transcription start site were isolated and assayed for their ability to control the expression of the EGP-2 cDNA, the green fluorescent protein, the luciferase reporter gene and the thymidine kinase and cytosine deaminase suicide genes. Expression of these chimeric constructs as assessed in a range of different cell lines was restricted to cell lines expressing EGP-2. In addition, only cells expressing EGP-2 were sensitive for gancyclovir after being transiently transfected with EGP-2 promoter-driven thymidine kinase. Deletion analyses defined 687 bp upstream as the basic proximal promoter region, which could confer epithelial-specific expression to the GFP reporter gene in vitro. As these EGP-2 sequences can confer promoter activity to reporter and suicide genes in an EGP-2 restricted manner, they may be useful for gene therapy of EGP-2 expressing carcinomas.

MeSH terms

  • Animals
  • Antigens, Neoplasm / genetics*
  • Antigens, Neoplasm / metabolism
  • Antiviral Agents / pharmacology
  • Base Sequence
  • Cell Adhesion Molecules / genetics*
  • Cell Adhesion Molecules / metabolism
  • Cells, Cultured
  • Cytosine Deaminase / metabolism
  • Epithelial Cell Adhesion Molecule
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism*
  • Epithelial Cells / pathology
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Ganciclovir / pharmacology
  • Gene Expression Regulation*
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Luciferases / metabolism
  • Mice
  • Molecular Sequence Data
  • Neoplasms / genetics
  • Neoplasms / metabolism*
  • Neoplasms / pathology
  • Promoter Regions, Genetic / genetics*
  • Recombinant Fusion Proteins / metabolism*
  • Sequence Deletion
  • Thymidine Kinase / metabolism
  • Transcription Initiation Site
  • Transfection

Substances

  • Antigens, Neoplasm
  • Antiviral Agents
  • Cell Adhesion Molecules
  • Epithelial Cell Adhesion Molecule
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Luciferases
  • Thymidine Kinase
  • Cytosine Deaminase
  • Ganciclovir