Application of Escherichia coli nitroreductase and granulocyte macrophage colony-stimulating factor for glioma gene therapy

• Main Authors: Chun-Chieh Yeh, 葉俊頡
• Other Authors: Chien-Kuo Tai
• Format: Others
• Language: en_US
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/19077890829651354226

碩士 === 國立中正大學 === 分子生物研究所 === 97 === Single prodrug-activation system does not often induce complete tumor regression or avoid tumor recurrence after termination of prodrug administration. This limitation has more recently led to the use of combined approaches in which prodrug activation schemes are combined with each other in a synergistic manner. In our study, we assessed the antitumor effect of combined gene therapy for brain tumors, using genes for Escherichia coli nitroreductase (NTR) together with granulocyte macrophage colony-stimulating factor (GM-CSF). We prove that expression of NTR in rat glioma cells (RG2 cells) enables activation of the prodrug CB1954, subsequently leading to cell death. Moreover, we used RG2 cells constitutively expressing NTR to demonstrate that the NTR/CB1954 system results in potent, long-lasting antitumoral effects in rats. Next, our findings indicated that two replicating retroviral vectors, ACE (modified from amphotropic murine leukemia virus, MLV) and GS (modified from Gibbon ape leukemia virus, GaLV) vectors carrying GFP and dsRed genes as markers could co-infected and express two report genes in RG2. When replacing reporter genes with NTR and mGMCSF, we tested the cell killing effect of vector expressing NTR upon administration of CB1954 and detected cytokine secretion of vector expressing mGMCSF. Our future experimental results my indicate that if combination of two replicating retroviral vectors with NTR and mGMCSF will produce greater therapeutic efficacy than NTR alone, providing effective strategy for brain tumor therapy.