Cancer cell-specific luciferase expression driven by the human survivin promoter
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: Survivin, a novel member of the inhibitor of apoptosis (IAP) protein family, is associated with malignant transformation and is over-expressed in oral squamous cell carcinoma (OSCC), but is undetectable in most normal adult tissues. We hypothesized that the survivin promoter could be employed to enhance the specificity of therapeutic gene expression in OSCC cells. We examined the activity of luciferase expressed from plasmids encoding the enzyme under the control of either the human survivin promoter (pSRVN.Luc) or the cytomegalovirus promoter (pCMV.Luc) in tumor- and non-tumor-derived epithelial cells. Methods: HeLa human cervical epithelial cancer cells, HSC-3, OSCC cells of the tongue, and GMSM-K, non-tumor-derived human oral epithelial cells, were seeded in 48-well culture plates one day before transfection, and used at ~80% confluence. The cells were transfected with two novel commercially available transfection reagents, the polycationic liposome, Metafectene (Biontex Laboratories GmbH, Munich, Germany), and the polyamine reagent, GeneJammer (Stratagene, La Jolla, CA) at optimized reagent:DNA ratios. Transfection efficacy was evaluated by measuring luciferase activity, using the Luciferase Assay System obtained from Promega and a Turner Designs TD-20/20 luminometer. The data were expressed as relative lights units (RLU) per ml of cell lysate. Results: All three cell lines displayed significant luciferase activity when transfected with pCMV.Luc. Both with Metafectene and GeneJammer, the survivin promoter-driven luciferase was expressed to high levels in HeLa and HSC-3 cells. Although GMSM-K cells expressed relatively high levels of luciferase driven by the CMV promoter, expression under the control of the survivin promoter was very low or not detectable. Conclusions: Our results suggest that the survivin promoter may be a promising tumor-specific promoter that may be used in gene therapy approaches for the treatment of oral cancer. This work was supported by funds from the University of the Pacific, Arthur A. Dugoni School of Dentistry.
Cancer cell-specific luciferase expression driven by the human survivin promoter.
Paper presented at The 35th Annual Meeting of the American Association for Dental Research in Orlando, FL.