Transcriptionally Targeted Nonviral Gene Transfer Using a β-Catenin/TCF-Dependent Promoter in a Series of Different Human Low Passage Colon Cancer Cells

• Published in: Molecular pharmaceutics Vol. 4; no. 1; pp. 129 - 139
• Main Authors: Gaedtke, Lars, Pelisek, Jaroslav, Lipinski, Kai S, Wrighton, Christopher J, Wagner, Ernst
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.01.2007
• Subjects: beta Catenin - genetics; 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
• ISSN: 1543-8384; 1543-8392
• DOI: 10.1021/mp0600586

Nonviral transfections of six low passage human colon cancer cell lines using the artificial β-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. Keywords: Wnt signaling pathway; β-catenin; colorectal cancer; nonviral gene therapy; lipopolyfection; multicellular spheroids