Development of Transferrin-Polycation/DNA Based Vectors for Gene Delivery to Melanoma Cells

Abstract We describe the comparison of non-viral polycation transfection reagents, adenovirus-enhanced transferrinfection (AVET), polyethylenimine (PEI800) and transferrin-conjugated PEI800 (Tf-PEI800) in their ability to transfect murine and primary human melanoma cell lines. Expression of a report...

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Bibliographic Details
Published in:Journal of drug targeting 1999-01, Vol.7 (4), p.293-303
Main Authors: Wightman, Lionel, Patzelt, Erik, Wagner, Ernst, Kircheis, Ralf
Format: Article
Language:English
Subjects:
Adenoviridae - chemistry
Animals
AVET
Cations - chemistry
Cell Line
Cytokines - metabolism
DNA - metabolism
Drug Delivery Systems - methods
Genes, Reporter - genetics
Genetic Vectors - genetics
Humans
Interleukin-2
Interleukin-2 - metabolism
Luciferases - genetics
Melanoma
Melanoma - metabolism
Membrane Proteins - metabolism
Mice
Polyethyleneimine - chemistry
Polyethylenimine
Transfection
Transfection - genetics
Transferrin
Transferrin - genetics
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Summary:Abstract We describe the comparison of non-viral polycation transfection reagents, adenovirus-enhanced transferrinfection (AVET), polyethylenimine (PEI800) and transferrin-conjugated PEI800 (Tf-PEI800) in their ability to transfect murine and primary human melanoma cell lines. Expression of a reporter gene, cell surface marker and secreted protein (interleukin-2) was assessed for each vector system. Testing for luciferase reporter gene expression in murine and primary human cell lines, AVET and Tf-PEI800, both showed high levels of expression and comparable activity. Furthermore, when the melanoma cell line B16F10 was transfected with a cell surface marker up to 97% of the cells expressed the protein on the cell surface. Assessing the levels of secreted IL-2 in murine cell lines, AVET/IL-2, Tf-PEI800/IL-2 and PEI800/IL-2 all expressed high levels of the cytokine (up to 20 μg IL-2/106 cells/24 h). In primary human melanoma cell lines, AVET/IL-2 transfected cells secreted more IL-2 than cells transfected with either Tf-PEI800/IL-2 or PEI800/IL-2. In murine melanoma cell culture experiments, positively charged PEI800/ DNA and Tf-PEI800/DNA complexes gave similar transfection efficiencies. However, when subcutaneous tumors in mice were injected with the luciferase reporter gene com-plexed with either Tf-PEI800 or AVET, higher transfection activity was measured in the tumors as compared to ligand free PEI800/DNA complexes.
ISSN:1061-186X
1029-2330
DOI:10.3109/10611869909085512