Enhancing gene delivery to cancer cells by transferrin and EGF
Nejat Düzgüneş: 0000-0001-6159-1391
87th General Session of the International Association for Dental Research (IADR) and 38th Annual Meeting of the American Association for Dental Research (AADR)
April 1-4, 2009
Date of Presentation
Journal of Dental Research
Journal Volume Number
88 (Special issue A)
Objectives: Despite advances in diagnosis and treatment of head and neck squamous cell carcinoma (HNSCC), including improvements in radiation therapy, surgical techniques, chemotherapy and prevention strategies, survival rates for patients with recurrent head and neck cancer are poor. Our laboratory is developing safe, non-viral vectors for the delivery of therapeutic genes to HNSCC cells. The purpose of this study was to increase transfection efficiency of the polycationic, non-viral vector Metafectene Pro (MP) via complexation of Transferrin or Epidermal Growth Factor (EGF) as targeting ligands, and to determine if the resistance of some HNSCC cells to transfection could be overcome by this method. Methods: HSC-3 and H-413 cells were seeded in 48-well culture plates the day before transfection, and used at approx. 85% confluence. The plasmid pCMV.Luc expressing luciferase under the control of the cytomegalovirus promoter was complexed with MP and with either human rEGF or human Transferrin. Luciferase activity was assayed 48 hours after transfection. Results: EGF- and Transferrin-mediated enhancement of transfection was most prominent under conditions where MP alone was sub-optimal for transfection (1 µl MP/µg DNA). In HSC-3 cells, complexation of Transferrin or EGF caused an approx. 7-fold increase in luciferase expression. In H-413 cells that are very resistant to transfection, complexation of EGF with MP lipoplexes resulted in a 15-fold increase in luciferase activity, while Transferrin mediated only a 2-fold enhancement. Use of 1.5 and 2 µl MP in H-413 cells enabled EGF to increase transfection by 12- and 4-fold, respectively. Conclusions: Transfection of HNSCC cell with lipoplexes can be improved with the use of Transferrin and EGF as targeting ligands, even in cells that are difficult to transfect. Such lipoplexes may be used for cancer cell-specific delivery of suicide genes in the therapy of HNSCC. Funded by Research Pilot Project Award 03-Activity 062.
Enhancing gene delivery to cancer cells by transferrin and EGF.
Paper presented at 87th General Session of the International Association for Dental Research (IADR) and 38th Annual Meeting of the American Association for Dental Research (AADR) in Miami, FL.