Transcriptionally targeted nonviral gene transfer using a beta-catenin/TCF-dependent promoter in a series of different human low passage colon cancer cells

Mol Pharm. Jan-Feb 2007;4(1):129-39. doi: 10.1021/mp0600586.

Abstract

Nonviral transfections of six low passage human colon cancer cell lines using the artificial beta-catenin/TCF-dependent promoter CTP4 demonstrated a high promoter activity which was 1000- to 70000-fold higher than in HeLa control cells. Luciferase gene expression levels obtained with CTP4 in epithelial-like tumor cell cultures were only slightly lower than with the strong viral CMV promoter/enhancer, whereas in less differentiated tumor cultures CTP4 expression levels exceeded the CMV expression levels up to 28-fold. Three cell lines representing different morphology typical of the original tumors, more differentiated epithelial-like (COGA-5), piled-up (COGA-12), and poorly differentiated rounded-up (COGA-3), were selected for further investigation. Gene transfer was optimized using lipopolyplex formulation of cationic lipid DOSPER and polycation PEI25br. Lipopolyplexes enabled up to 1300-fold or 400-fold higher luciferase expression compared to the corresponding lipoplexes or polyplexes, respectively. Lipopolyfection of an interleukin-2 (IL-2) gene expression construct driven by the CTP4 promoter resulted in very high levels of up to 95 ng of secreted IL-2 per 105 cells and 24 h. The lipopolyplexes were also able to transfect multicellular spheroids that mimic the three-dimensional structure of real tumors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Colonic Neoplasms / pathology*
  • Cytomegalovirus
  • DNA / metabolism
  • Green Fluorescent Proteins / metabolism
  • HeLa Cells
  • Humans
  • Interleukin-2 / biosynthesis
  • Lipids
  • Luciferases / metabolism
  • Plasmids / metabolism
  • Promoter Regions, Genetic / genetics*
  • Spheroids, Cellular / cytology
  • TCF Transcription Factors / metabolism*
  • Transcription, Genetic*
  • Transfection / methods*
  • Tumor Cells, Cultured
  • beta Catenin / genetics*

Substances

  • Interleukin-2
  • Lipids
  • TCF Transcription Factors
  • beta Catenin
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • DNA
  • Luciferases