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  • Title: [Effects of CD/5-FC suicide gene therapy system on human malignant glioma cells in vitro].
    Author: Lü SQ, Liu YS, Yang H.
    Journal: Zhong Nan Da Xue Xue Bao Yi Xue Ban; 2004 Apr; 29(2):174-6, 180. PubMed ID: 16145905.
    Abstract:
    OBJECTIVE: To investigate the killing effects of cytosine deaminase/5-fluorocytosine (CD/5-FC) suicide gene therapy system on human malignant glioma cells in vitro. METHODS: The pCMVCD plasmid was transfected into the U251 human malignant glioma cells using Lipofectamine 2000-mediated method. Resistant clones (named U251/CD cells) were isolated by screening with G418 presence. U251/CD cells were incubated with 5-FC in different concentrations to determine the number of viability cells (or cytotoxicity assay), and measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The concentrations of 5-fluorouracil (5-FU) in the media were measured by the high-performance liquid chromatography (HPLC) analysis. RESULTS: The untreated U251 cells were insensitive to 5-FC, with the IC50 about 6 500 micromol/L. After the transfection, the IC50 was dramatically reduced to about 10 micromol/L. Therefore, gene transfection made G418-resistant clones (U251/CD cells) highly sensitive to 5-FC. HPLC analysis showed that 5-FU was detected in U251/CD cell medium. CONCLUSION: The killing effects of U251 cells modified by CD gene were induced by 5-FC, and the CD/5-FC system was feasible to treat glioma.
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