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| Killing effects of cytosine deaminase gene mediated by adenovirus vector on human pancreatic cancer cell lines in vitro |
| Zhao-Shen Li, Xue Pan, Guo-Ming Xu, Long Cui, Guan-Rong Dai, Yan-Fang Gong and Zhen-Xing Tu |
Shanghai, China
From the Department of Gastroenterology (Li ZS, Pan X and Xu GM); Department of General Surgery (Cui L, Dai GR, Gong YF and Tu ZX), Changhai Hospital, Second Military Medical University, Shanghai 200433, China
Correspondence: Zhao-Shen Li, MD (Tel: 86-21-250705 52; Fax: 86-21-55621735; Email: zsli@online.sh.cn) |
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Abstract OBJECTIVE: To evaluate the killing effects of the cytosine deaminase (CD) gene mediated by adenovirus vector on human pancreatic cancer cell lines in vitro.
METHODS: The CD gene was cloned into pAdTrack-CMV-CD, and pAdTrack-CMV-CD and pAdEasy-1 were recombinated in bacteria. The newly recombinated Ad-CD containing green fluorescent protein (GFP) was propagated in 293 cells and purified by cesium chloride gradient centrifugation. Human pancreatic cancer cell lines Patu8988 and SW1990 were infected with this virus, then 5-FC was added. XTT assay was used to estimate relative numbers of viable cells.
RESULTS: The positive clones were selected by using endonuclease to digest the combinatants and the concentration of viral liquids containing the CD gene was 2×1011 pfu/ml. It was found that significant cytotoxic activities were possessed by 5-FC for the CD gene transduced pancreatic cell lines, but little effects exerted on the nontransduced pancreatic carcinoma cells.
CONCLUSIONS: The CD gene mediated by adenovirus with a high infectivity is efficient for gene therapy of pancreatic carcinoma cell lines. These data demonstrate the therapeutic efficacy of an enzyme prodrug strategy in experimental pancreatic cancer.
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2003, Vol. 2 
