Killing effects of cytosine deaminase gene mediated by adenovirus vector on human pancreatic cancer cell lines in vitro

<正> OBJECTIVE: To evaluate the killing effects of the cytosine deaminase （CD） gene mediated byadenovirus 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-lwere recombinated in bacteria. The newly...

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Bibliographic Details
Published in:Hepatobiliary & pancreatic diseases international 2003-02, Vol.2 (1), p.147-151
Main Authors: Li, Zhao-Shen, Pan, Xue, Xu, Guo-Ming, Cui, Long, Dai, Guan-Rong, Gong, Yan-Fang, Tu, Zhen-Xing
Format: Article
Language:English
Subjects:
Adenoviridae - genetics
adenovirus
cancer
Cell Line, Tumor
Cloning, Organism - methods
cytosine
Cytosine Deaminase - genetics
deaminase
gene
Genetic Therapy - methods
Genetic Vectors - genetics
Humans
pancreatic
Pancreatic Neoplasms - genetics
Pancreatic Neoplasms - therapy
Prodrugs - therapeutic use
therapy
Treatment Outcome
verctor
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Summary:<正> OBJECTIVE: To evaluate the killing effects of the cytosine deaminase （CD） gene mediated byadenovirus 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-lwere recombinated in bacteria. The newly recombinated Ad-CD containing green fluoreseent protein（GFP） was propagated in 293 cells and purified by cesium chloride gradient centrifugation. Humanpancreatic 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 theconcentration of viral liquids containing the CD gene was 2×1O11 pfu/ml. It was found that significantcytotoxic activities were possesscd by 5-FC for the CD gene transduced pancreatic cell lines, but littleeffects exerted on the nontransduced pancreatic carcinoma cells.CONCLUSIONS: The CD gene mediated by adenovirus with a high infectivity is efficient for genetherapy of pancreatic carcinoma cell lines. These data demonstrate the therapeutic efficacy of an enzymeprodrug strategy in experimental pancreatic cancer.
ISSN:1499-3872