Killing effects of cytosine deaminase gene mediated by adenovirus vector on human pancreatic cancer cell lines in vitro / 中华消化杂志

ABSTRACT

Objective To evaluate the killing effects of cytosine deaminase (CD) gene mediated by adeno virus vector on human pancreatic cancer cell lines in vitro. Methods After CD gene was cloned into pAdTrack CMV CD, 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 Patu 8988 and SW 1990 were infected with this virus, then 5 FC was added, XTT assay was used to estimate relative numbers of viable cell. Results The positive clones were selected by using endonuclease to digest the combinatants and the concentration of viral liquids containing CD gene was 2?10 11 PFU/ml. It was found that significant cytotoxic activities were possessed by 5 FC for CD gene transduced pancreatic cell lines, but little effects on the nontransduced pancreatic carcinoma cells. Conclusions CD gene mediated by adenovirus has a high infectivity and is efficient for gene therapy of pancreatic carcinoma cell lines. These data demonstrate the therapeutic efficacy of an enzyme as a prodrug strategy in experimental pancreatic cancer.