Combining Gene Therapy with Adenoviral Vector Encoding 4-1BB L, B7-1 and p53 in Skov3 Cell Lines

• Published in: Cell journal (Yakhteh) 2011-01, Vol.12, p.21-21
• Main Authors: Hosseini, A, Amirmoezi, F, Jaberipour, M, Habibagahi, M
• Format: Article
• Language: English
• Subjects: Adenovirus; Apoptosis; B7-1 antigen; Blood; Cancer; CD4 antigen; CD8 antigen; CD80 antigen; Cell proliferation; Cell survival; Expression vectors; FasL protein; Flow cytometry; gamma -Interferon; Gene therapy; Infection; Lymphocytes; p53 protein; Polymerase chain reaction; Solid tumors; Titration; Tumor cells
• ISSN: 2228-5806

Objective: Combination gene therapy is an important approach in cancer therapy. Conventional therapy snowed significant result to cancer treatment. Previously, it was showed that co-stimulation with 4-IBB ligand (CD137L) and B7-1 (CD80) can trigger lymphocytes to kill tumor cells but some of these cells can escape from stimulated lymphocytes. Furthermore, replacement mutant p53 with wild type p53 demonstrate significant result in some solid tumors. In this study, we investigated the efficacy of combined adenoviral gene transfer of CD80, CD 137L and p53 into Skov3 cell lines to prevent cancer progression and in vitro elimination cancer cells. Materials and Methods: Adenoviral vector that encoding CD80, CD137L and wild type p53 was constructed. After virus propagation and titration, the titer of CD80 and CD137L adenovirus was calculated to induce desired expression. Also, IC 50 of wild type p53 was measured. Skov3 cells were infected and after 48 hours were co-cultured with CFSE pre-stained PBMCs that separated from blood of healthy volunteer. 3 days after co-culture, lymphocytes were harvested and lymphocyte proliferation and IFN- gamma expression was examined in CD4+ and CD8+ lymphocytes using FACS analyzing. Expression of FasL and granzymeB was determined using Real-Time PCR. Apoptosis induction and survival of tumor cells were evaluated using MTT assay. Results: Result of this study showed that more than 90% of Skov3 cells expressed CD80 and CD137L after infection by 600 virus particle per cells (VP/Cell). 1000 VP/Cell of P53 adenovirus induced cell death in 50% of Skov3 cells. After 3 days co-cultured infected cells with PBMCs, lymphocyte proliferation in dual co-stimulation and triple condition was statistically increased in compare to GFP infected cells. IFN- gamma was more expressed in dual co-stimulation and triple condition in both CD4+ and CD8+ population (p0.05). Interestingly, less than 10% of infected cells with triple viruses were survived while 50% and 60% of cells infected with p53 and dual co-stimulation were survived, respectively. Conclusion: Our results showed that although dual co-stimulation can trigger lymphocytes to kill tumor cells, wild type p53 beside to dual co-stimulation have profound effects in tumor cell killing.