|2394 Murine Survivin Promoter-Driven Gene Expression in Cancer and Non-Tumor Cells|
C. SPAIN1, N. OVERLID1, F. LI2, N. DÜZGÜNES1, and K. KONOPKA1, 1University of the Pacific, Arthur A. Dugoni School of Dentistry, San Francisco, CA, USA, 2Roswell Park Cancer Institute, Buffalo, NY, USA|
Objectives: Survivin, a novel member of the inhibitor of apoptosis protein family, is over-expressed in oral squamous cell carcinoma (OSCC). We have hypothesized that this property may be exploited for the expression of therapeutic genes specifically in oral cancer cells. We examined the activity of reporter gene, luciferase, expressed from plasmids encoding the enzyme under the control of either the cytomegalovirus promoter (pCMV.Luc), or the murine (pSRVN.Luc-1342 and pSRVN.Luc-194) or the human (pSRVN.Luc-1430) survivin promoter, in tumor and non-tumor cells. Methods: Murine squamous cell carcinoma SCCVII cells, NIH-3T3 murine fibroblasts, and murine mammary gland NMuMG cells were transfected with a novel polycationic liposome, Metafectene PRO at a ratio of 2µl :1 µg DNA. Transfection efficacy was evaluated by measuring luciferase activity, using the Luciferase Assay System and a Turner Designs TD-20/20 luminometer. The data were expressed as relative light unites (RLU) per ml of cell lysate or per mg of protein. Results: All three cell lines displayed significant luciferase activity when transfected with pCMV.Luc. The expression of luciferase under the control of the murine and human survivin promoters was much lower than that obtained with the CMV promoter. In SCCVII cells, luciferase activities expressed under the murine pSRVN.Luc-1342 and pSRVN.Luc-194 promoters were ~3600-fold and ~7800-fold less than that obtained with pCMV.Luc. Gene expression from the survivin promoters in SCCVII cells was not significantly different than in NIH-3T3 cells. Conclusions: Gene expression from the murine and human survivin promoters in SCCVII cancer cells and non-tumor murine cells is much lower than that from the CMV promoter. The results indicate that survivin promoters do not significantly enhance gene expression in murine cancer cells vs. normal cells. This work was partially supported by Research Pilot Project Award 03-Activity 042 from the University of the Pacific, Arthur A. Dugoni School of Dentistry (K. Konopka).
|Seq #240 - Premalignant Oral Lesions and Oral Cancer|
3:30 PM-4:45 PM, Friday, March 23, 2007 Ernest N. Morial Convention Center Exhibit Hall I2-J