Abstract
Loss of p53 gene function, which occurs in most colon cancer cells, has been shown to abolish the apoptotic response to 5-fluorouracil (5-FU). To identify genes downstream of p53 that might mediate these effects, we assessed global patterns of gene expression following 5-FU treatment of isogenic cells differing only in their p53 status. The gene encoding mitochondrial ferredoxin reductase (protein, FR; gene, FDXR) was one of the few genes significantly induced by p53 after 5-FU treatment. The FR protein was localized to mitochondria and suppressed the growth of colon cancer cells when over-expressed. Targeted disruption of the FDXR gene in human colon cancer cells showed that it was essential for viability, and partial disruption of the gene resulted in decreased sensitivity to 5-FU-induced apoptosis. These data, coupled with the effects of pharmacologic inhibitors of reactive oxygen species, indicate that FR contributes to p53-mediated apoptosis through the generation of oxidative stress in mitochondria.
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Acknowledgements
We thank L. Meszler for help with cell imaging and all members of the Kinzler/Vogelstein Laboratories for advice and discussion. This work was supported by the Clayton Fund, the Miracle Foundation, and NIH grants CA 43460 and GM 07184. K.W.K. receives research funding from Genzyme Molecular Oncology (Genzyme) and K.W.K. and B.V. are consultants to Genzyme. Under a licensing agreement between the Johns Hopkins University and Genzyme, the SAGE technology was licensed to Genzyme, and K.W.K. and B.V. are entitled to a share of royalty received by the University from sales of the licensed technology. The terms of these arrangements are being managed by the University in accordance with its conflict of interest policies.
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Supplemental Figure A
a, p53 western blot of parental HCT116 (WT), TRP53-/-, FDXR+/+/- (G10, A11) and FDXR+/-/- (C2, D2) cell lines treated (+) with 50 mg/ml 5-FU for 48 h versus untreated (-) samples. b, p21 western blot of parental HCT116 (WT), FDXR+/+/- (G10), FDXR+/-/- (D2, C2) cell lines treated with 0 (for WT only, same baseline for all samples), 30 or 50 mg/ml 5-FU for 48 h. Equal amounts of protein (25 mg) were loaded in each well. (JPG 27 kb)
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Hwang, P., Bunz, F., Yu, J. et al. Ferredoxin reductase affects p53-dependent, 5-fluorouracil–induced apoptosis in colorectal cancer cells. Nat Med 7, 1111–1117 (2001). https://doi.org/10.1038/nm1001-1111
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DOI: https://doi.org/10.1038/nm1001-1111
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