Adenovirus-mediated gene transfer in the treatment of pancreatic cancer

Introduction: Suicide gene therapy is a new experimental form of cancer chemotherapy that is currently being evaluated in human trials. Aim: To evaluate the killing effects of the cytosine deaminase (CD) gene mediated by an adenovirus vector on human pancreatic carcinoma in vitro and in vivo. Methodology: The CD gene was cloned into pAdTrack-CMV-CD, pAdTrack-CMV-CD, and pAdEasy-1, which underwent recombination in bacteria BJ5183. The newly recombinant Ad-CD containing green fluorescent protein was propagated in 293 cells and purified by cesium chloride gradient centrifugation. The human pancreatic carcinoma cell line PaTu8988/SW1990 was infected with this virus, and then 5-fluorocytosine (5-FC) was added; XTT assay was used to estimate relative numbers of viable cells. An in vivo model of pancreatic cancer was established by injecting 1.0 x 10(7) PaTu8988/SW1990 cells subcutaneously in Balb/c nude mice. When tumors were palpable, Ad-CD was injected into each tumor, and 5-FC was administered. The positive clones were selected by endonuclease digestion of the combinations, and the concentration of viral liquids containing the CD gene was 2 x 10(11) pfu/mL. Results: It was found that significant cytotoxic activities were possessed by 5-FC for CD gene-transduced PaTu8988/SW1990 cells, but there was little effect on the nontransduced pancreatic carcinoma cells. The antitumor effect was observed in PaTu8988/SW1990 xenografts from nude mice with in situ CD gene transduction. Conclusion: These results indicate that the CD gene mediated by adenovirus has a high level of infectivity and is efficient for gene therapy for pancreatic carcinoma.