Gene Therapy For Human Ovarian Cancer With Highly Efficient And Targeted Expression Of Fas Gene

Fas gene is one of the most important genes to induce apoptosis. The abnormality of Fas/FasL pathway not only relates to the genesis and progression of tumors, it also has links to the chemosensitivity of some anticancer drugs. The loss or low expression of fas gene in some ovarian cancer cells results in the insensitivity of inducing apoptosis as well as the resistance of chemotherapy for ovarian cancer cells. It is reported that transduction of Fas gene into tumor cells may be a promising method of gene therapy for ovarian cancer. One of the major obstacles of gene therapy is that the ectopic gene cannot express highly in targeted cells. The objective of this study focuses on how to guarantee Fas gene expresses highly in tumor site.In the first part of this study, flow cytometry was used to assess the different ovarian cancer cell lines expression of Fas. We also used the adenoviral vector Ad5 and chimeric adenoviral vector Ad5F35 carried a report gene (enhanced green fluorescent protein gene, EGFP) to detect the infective efficacy for different ovarian cancer cell lines. The results showed that expression of Fas in tumor cells appears heterogeneity, and the expression level of Fas in SKOV3 cell line is very low. The infective efficacies of adenoviruses to ovarian cancer cells have great difference; but for the same cell line, Ad5 has higher infective efficacy than Ad5F35. In the second part, we constructed three shuttle plasmids contained different control elements of hTERT promoter and two-step transcriptional amplification(TSTA) system, they were named as pDC316-hTERT-Fas, pDC316...