Serum-resistant gene delivery to murine squamous cell carcinoma cells
Nejat Düzgüneş: 0000-0001-6159-1391
The 35th Annual Meeting of the American Association for Dental Research
March 8-12, 2006
Date of Presentation
Journal of Dental Research
Journal Volume Number
85 (Special issue A)
Objectives: One of the genetic approaches to the treatment of head and neck squamous cell carcinoma (HNSCC) is based on the hypothesis that expression of therapeutic genes in target cells will cause cytotoxicity or apoptosis. Gene delivery by non-viral vectors is usually inhibited by biological milieu. We examined whether two novel transfection reagents could introduce reporter and therapeutic genes in the presence of mouse serum. Methods: Transfection activity of the polycationic liposome, Metafectene, and the polyamine reagent, GeneJammer, was evaluated by measuring luciferase activity in murine squamous cell carcinoma cells, SCCVII, after a 48-hour incubation. Transfection efficiency was estimated by ß-galactosidase staining. Cells transfected with the Herpes Simplex Virus thymidine kinase (HSV-tk) plasmid were incubated in the absence or presence of ganciclovir (GCV; 20 µg/ml). Mock-transfected cells served as controls. The Alamar Blue assay was used to determine GCV-specific cytotoxicity. Results: Metafectene-mediated luciferase expression was reduced by 70% in 20% serum and 25% in 60% serum. GeneJammer-mediated luciferase expression was reduced by 45-55% in the presence of 20-60% serum. The expression of ß-galactosidase was essentially not affected by serum. Approx. 50% of the cells was positive for ß-gal staining. The delivery of the HSV-tk gene by Metafectene in the presence of 0% or 60% serum, followed by GCV treatment for 7 days, resulted in 90% and 82% cytotoxicity in 0% and 60% mouse serum, respectively. With GeneJammer, after 6 days, 95% cytotoxicity was observed both in 0% and 60% serum. Conclusions: Our observations suggest that Metafectene and GeneJammer may be useful for the gene therapy of OSCC in biological milieu. This work was supported by funds from the University of the Pacific, Arthur A. Dugoni School of Dentistry.
Serum-resistant gene delivery to murine squamous cell carcinoma cells.
Paper presented at The 35th Annual Meeting of the American Association for Dental Research in Orlando, FL.