Serum-resistant gene delivery to murine squamous cell carcinoma cells

ORCiD

Nejat Düzgüneş: 0000-0001-6159-1391

Department

Biomedical Sciences

Document Type

Conference Presentation

Conference Title

The 35th Annual Meeting of the American Association for Dental Research

Location

Orlando, FL

Conference Dates

March 8-12, 2006

Date of Presentation

3-10-2006

Journal Title

Journal of Dental Research

Journal ISSN

0022-0345

Journal Volume Number

85 (Special issue A)

First Page

859

Abstract

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.

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