Serum-resistant HSVtk/ganciclovir gene therapy in oral cancer cells


Nejat Düzgüneş: 0000-0001-6159-1391


Biomedical Sciences

Document Type

Conference Presentation

Conference Title

82nd General Session of the International Association for Dental Research and 33rd Annual Meeting of the American Association for Dental Research


Honolulu, HI

Conference Dates

March 10-13, 2004

Date of Presentation


Journal Title

Journal of Dental Research

Journal ISSN


Journal Volume Number


First Page



Objectives: Oral Squamous Cell carcinoma (OSCC) is the most prevalent cancer involving the oral cavity and oropharynx. The purpose of this study was to deliver the Herpes Simplex Virus thymidine kinase gene (HSV-tk) to HSC-3 human SCC cells, as a “suicide” gene therapy approach. We examined the effect of fetal bovine serum (FBS) on the delivery of a reporter gene and HSV-tk by the polyamine reagent, Gene Jammer, and the polycationic liposome, Metafectene. Methods: We assessed gene delivery by incubating HSC-3 cells with the plasmid pCMV-luc and measuring luciferase activity in cell lysates. Cells transfected with the HSV-tk plasmid were incubated in the absence and the presence of ganciclovir (10 µg/ml) for the indicated periods of time. We used the Alamar Blue assay to determine ganciclovir-specific cytotoxicity to HSC-3 cells mediated by the plasmid pCMV-HSVtk. Mock-transfected cells served as controls. Results: The optimal ratios of the reagents to DNA for delivering the luciferase gene were 4 ml Metafectene:µg DNA and 6 or 12 ml GeneJammer:µg DNA. GeneJammer-mediated luciferase expression was inhibited by about 30% when transfection was performed in the presence of 10% FBS. The delivery of the HSV-tk gene by Gene Jammer in the absence and presence of 10% FBS, followed by ganciclovir treatment for 9 days, resulted in 100% and 70% cytotoxicity, respectively. With Metafectene, FBS in the range 10-60% inhibited luciferase gene expression by about 60%. Using HSVtk/ganciclovir, 90-100% cytotoxicity was observed in the presence of 0 or 10% FBS. Even in the presence of 60% FBS, Metafectene mediated 50-70% cytotoxicity, compared to controls. Conclusions: Our observations suggest that Metafectene may be useful for the gene therapy of OSCC in animal models. (This work was supported partially by Research Award DRES03-027 from the School of Dentistry).

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