p14ARF upregulation of p53 and enhanced effects of 5-fluorouracil in pancreatic cancer.
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.


CONCLUSIONS
Combined use of p14ARF gene and 5-Fu acts synergistically to inhibit pancreatic cancer cell proliferation, suggesting a new anticancer strategy.
Zhang
Qunhua
Q
Center for Pancreatic Cancer, Huashan Hospital, Fudan University, Shanghai 200040, China. qunhuazh@sh163a.sta.net.cn
Ni
Quanxing
Q
Gan
Jun
J
Shen
Zhaozhong
Z
Luo
Jianmin
J
Jin
Chen
C
Zhang
Niu
N
Zhang
Yanling
Y
D005472:Fluorouracil / Q000494:pharmacology*; D006801:Humans; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000628:therapy; D014162:Transfection; D014407:Tumor Cells, Cultured; D025803:Tumor Suppressor Protein p14ARF / Q000235:genetics*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*; D015854:Up-Regulation / Q000502:physiology*
D025803:Tumor Suppressor Protein p14ARF; D016159:Tumor Suppressor Protein p53; D005472:Fluorouracil