Objective To investigate the synergistic antitumor effects of combined use of p14ARF gene and 5-fluorouracil (5-Fu) in pancreatic cancer.Methods A human pancreatic cancer cell line PC-3 was transfected with lipofectin-mediated recombinant p14ARF gene,and was then administered with 5-Fu. Cell growth,morphological changes,cell cycle,apoptosis,and molecular changes were measured using the MTT assay,flow cytometry,RT-PCR,Western blotting,and immunocytochemical assays.Results After transfection of p14ARF,cell growth was obviously inhibited,resulting in an accumulation of cells in the G 1 phase. The proportion of cells in the G 1 phase was significantly increased from 58.51% to 75.92 %,and in the S and G 2/M phases decreased significantly from 20.05% to 12.60%,and from 21.44% to 11.48 %,respectively,as compared with those of the control groups. PC-3/p14ARF cells that underwent 5-Fu treatment had significantly greater G 2/M phase accumulation,from 11.48% to 53.47 %. The apoptopic index was increased in PC-3/p14ARF cells from 3.64% to 19.62%. The MTT assay showed p14ARF-expressing cells were significantly more sensitive to 5-Fu (0.01-10 mg/L) than those devoid of p14ARF expression ( P <0.01). Western blotting showed p14ARF upregulates p53 expression. Conclusion Combined use of p14ARF gene and 5-Fu acts synergistically to inhibit pancreatic cancer cell proliferation,suggesting a new anticancer strategy. Objective To investigate the synergistic antitumor effects of combined use of p14ARF gene and 5-fluorouracil (5-Fu) in pancreatic cancer. Methods A human pancreatic cancer cell line PC-3 was transfected with lipofectin-mediated recombinant p14ARF gene, and was then administered with 5-Fu. Cell growth, morphological changes, cell cycle, apoptosis, and molecular changes were measured using the MTT assay, flow cytometry, RT- PCR, Western blotting, and immunocytochemical assays. Results after transfection of p14ARF, cell growth was obviously inhibited, resulting in an accumulation of cells in the G 1 phase. The proportion of cells in the G 1 phase was significantly increased from 58.51% to 75.92%, and in the S and G 2 / M phases decreased significantly from 20.05% to 12.60 %, and from 21.44% to 11.48%, respectively, as compared to those of the control groups. PC-3 / p14ARF cells that underwent 5-Fu treatment had significantly greater G2 / M phase accumulation, from 11.48% to 53.47%. The apoptopic index The MTT assay showed p14ARF-expressing cells were significantly more sensitive to 5-Fu (0.01-10 mg / L) than those devoid of p14ARF expression (P <0.01 ). Western blotting showed p14ARF upregulates p53 expression. Conclusion Combined use of p14ARF gene and 5-Fu acts synergistically to inhibit pancreatic cancer cell proliferation, suggesting a new anticancer strategy.