Transfection of HeLa, HEK293 and HSC-3 cells by transferrin lipoplexes
Nejat Düzgüneş: 0000-0001-6159-1391
31st Annual Meeting of the American Association for Dental Research
San Diego, CA
March 6-9, 2002
Date of Presentation
Journal of Dental Research
Journal Volume Number
Objectives: Cationic lipid-DNA complexes (“lipoplexes”) are being used as a delivery system for therapeutic genes both in vitro and in vivo. The limitations of lipoplexes include their low transfection efficiency, their potential cytotoxicity at high concentrations and the inhibition of transfection by serum. Previous studies have indicated that the efficiency of transfection can be enhanced by complexation of human transferrin (Tf) with cationic liposomes before lipoplex formation. Tf is an iron-transporting serum glycprotein, which binds to the receptor expressed on the surface of proliferating cells with a particular high expression on erythroblasts and tumor cells. Methods: Here we examined the effect of Tf on DOTAP/DOPE (ESCORTTM) – mediated transfection of three different cell lines at various lipid/DNA (+/-) charge ratios, using the plasmid pCMVluc, which expresses luciferase as a reporter gene as described in Simoes et al. (Gene Ther 1998,5:955-964) Results: In HEK293 (human embryonal kidney), and HeLa (human cervical carcinoma) cells, the greatest enhancement of gene expression by Tf was achieved at a charge ratio of 1.0 with a 3-fold enhancement over plain lipoplexes. Tf-mediated enhancement of transfection in HSC-3 (human squamous cell carcinoma) cells was higher than observed with Hela and HEK293 cells. A 10-fold enhancement was obtained at a charge ratio of 0.5. The level of gene expression was highly dependent of the cell type, with HSC-3 showing the lowest luciferase activity, and HEK293 cells the highest. Conclusions: Complexation of Tf with cationic liposomes enhances transgene expression in a number of cell lines, including HSC-3 cells which are difficult to transfect with plain lipoplexes. Enhancement of gene delivery by Tf has potential applications to the delivery of suicide or therapeutic gene into oral cancer cells.
Transfection of HeLa, HEK293 and HSC-3 cells by transferrin lipoplexes.
Paper presented at 31st Annual Meeting of the American Association for Dental Research in San Diego, CA.