Transcriptionally targeted nonviral gene transfer using a beta-catenin/TCF-dependent promoter in a series of different human low passage colon cancer cells.
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.
Gaedtke
Lars
L
Department of Pharmacy, Center of Drug Research, Pharmaceutical Biology-Biotechnology, Ludwig Maximilian University, Munich, Germany.
Pelisek
Jaroslav
J
Lipinski
Kai S
KS
Wrighton
Christopher J
CJ
Wagner
Ernst
E
D003110:Colonic Neoplasms / Q000473:pathology*; D003587:Cytomegalovirus; D004247:DNA / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis; D008055:Lipids; D008156:Luciferases / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D018874:Spheroids, Cellular / Q000166:cytology; D051820:TCF Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D051176:beta Catenin / Q000235:genetics*
D007376:Interleukin-2; D008055:Lipids; D051820:TCF Transcription Factors; D051176:beta Catenin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004247:DNA; D008156:Luciferases