Abstract | BACKGROUND: MATERIAL/METHODS: The eukaryotic expression plasmid pCMV-CD was transfected into U251 malignant glioma cells. Resistant clones (labeled U251/CD cells) were subsequently isolated and further confirmed by reverse transcription polymerase chain reaction (RT-PCR), immunofluoroscence, and immunoblot. Then U251/CD cells were incubated with 5-FC at various concentrations to measure viability ratios as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. 5-FU concentrations in the media were measured by high-performance liquid chromatography (HPLC). Finally, the volumes and weights of tumors from glioma-bearing nude mice after 5-FC intervention were evaluated. RESULTS: The results revealed that the untreated U251 cells were insensitive to 5-FC whereas the U251/CD cells were highly sensitive. Apoptosis and cell death were observed on the U251/CD cells after 5-FC administration. HPLC analysis showed that 5-FU was detected in the U251/CD cell media. These in vivo animal data showed that the volumes and weights of the implanted tumors were dramatically decreased due to cell apoptosis and tumor necrosis. CONCLUSIONS: The in vivo results encourage a further investigation in a controlled trial on the treatment of malignant gliomas via the CD/5-FC gene therapy system.
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Authors | Sheng-Qing Lv, Ke-Bin Zhang, Eric E Zhang, Fang-You Gao, Chang-Lin Yin, Chun-Ji Huang, Jia-Quan He, Hui Yang |
Journal | Medical science monitor : international medical journal of experimental and clinical research
(Med Sci Monit)
Vol. 15
Issue 1
Pg. BR13-20
(Jan 2009)
ISSN: 1643-3750 [Electronic] United States |
PMID | 19114960
(Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- DNA Primers
- Tetrazolium Salts
- Thiazoles
- Flucytosine
- Cytosine Deaminase
- thiazolyl blue
- Fluorouracil
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Topics |
- Animals
- Cell Line, Tumor
- Chromatography, High Pressure Liquid
- Cytosine Deaminase
(genetics, metabolism)
- DNA Primers
(genetics)
- Escherichia coli
- Flucytosine
(metabolism)
- Fluorescent Antibody Technique
- Fluorouracil
(metabolism, therapeutic use)
- Genetic Therapy
(methods)
- Glioma
(drug therapy, genetics, therapy)
- Immunoblotting
- Mice
- Mice, Nude
- Reverse Transcriptase Polymerase Chain Reaction
- Tetrazolium Salts
- Thiazoles
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