Effects Of Fcy::Fur/5-FC Gene Therapy For Human Ovarian Cancer Cells

Objective:To detect the expression of Fcy::Fur suicide gene in eukaryotic cells, and to investigate the skilling effects of Fcy::Fur suicide gene therapy system on human ovarian cancer cells (SKOV3). To lay the foundation for the tumorous gene therapy with Fcy::Fur suicide gene.Methods: (1) With the technology of gene re-arrangement,Fcy::Fur gene in pORF -Fcy::Fur plasmid was subcloned into pEGFP-N1 vector , with its correctness evaluated by the means of restriction enzyme analysis and sequencing.(2)It was transfected into SKOV3 cells with lipofectin . The Stable transfectants were screened and were analyzed by RT-PCR and Western blot. (3) The killing efficacy of 5-FC on the gene transfered cells was determined by MTT and FCM assays.Results: (1) Correct construction of pEGFP-N1-Fcy::Fur was identified by methods of nucleotide sequence determination; (2) 60% transfected cells emitted out green fluorescence after 24 hour after transfection;(3) The transfected cells expressed Fcy::Fur gene by RT-PCR and western blot;(4) The MTT and FCM assays showed 5-FC killed SKOV3- Fcy::Fur cell. Conclusion: Fusion suicide gene therapy system Fcy::Fur/5-FC has a significant efficacy on human ovarian cancer cell SKOV3.