title: Increased Tumor Ascorbate is Associated with Extended Disease-Free Survival and Decreased Hypoxia-Inducible Factor-1 Activation in Human Colorectal Cancer.
abstract: Ascorbate is a co-factor for the hydroxylases that regulate the transcription factor hypoxia-inducible factor (HIF)-1, which provides cancer cells with a metabolic and survival advantage in the hypoxic environment of solid tumors. However, whether ascorbate affects tumor development is a highly debated issue. We aimed to determine whether tumor ascorbate was associated with HIF-1 activation and patient disease-free survival. In this study, we undertook a retrospective observational analysis of tissue-banked tumor and paired normal tissue from 49 colorectal cancer patients, measuring ascorbate levels, HIF-1α and its downstream gene products BNIP3, and vascular endothelial cell growth factor (VEGF). Patient survival was monitored for the first 6 years after surgery. We found that ascorbate levels were lower in tumor tissue compared to normal tissue (p < 0.001) but overall levels varied considerably. HIF-1α, VEGF, and BNIP3 were elevated in tumor samples (p < 0.01). There was an inverse relationship between tumor ascorbate content and HIF-1 pathway activation (p = 0.002) and tumor size (p = 0.018). Higher tumor ascorbate content was associated with significantly improved disease-free survival in the first 6 years after surgery (p = 0.006), with 141-1,094 additional disease-free days. This was independent of tumor grade and stage. Survival advantage was associated with the amount of ascorbate in the tumor, but not with the amount in adjacent normal tissue. Our results demonstrate that higher tumor ascorbate content is associated with decreased HIF-1 activation, most likely due to the co-factor activity of ascorbate for the regulatory HIF hydroxylases. Our findings support the need for future studies to determine whether raising tumor ascorbate is possible with clinical intervention and whether this results in modification of hydroxylase-dependent pathways in the tumor.
authors.affiliation: Centre for Free Radical Research, Department of Pathology and Biomedical Science, University of Otago Christchurch , Christchurch , New Zealand.
authors.forename: Caroline
authors.initials: C
authors.lastname: Kuiper
authors.affiliation: Mackenzie Cancer Research Group, Department of Pathology and Biomedical Science, University of Otago Christchurch , Christchurch , New Zealand.
authors.forename: Gabi U
authors.initials: GU
authors.lastname: Dachs
authors.affiliation: Mackenzie Cancer Research Group, Department of Pathology and Biomedical Science, University of Otago Christchurch , Christchurch , New Zealand.
authors.forename: Delwyn
authors.initials: D
authors.lastname: Munn
authors.affiliation: Mackenzie Cancer Research Group, Department of Pathology and Biomedical Science, University of Otago Christchurch , Christchurch , New Zealand.
authors.forename: Margaret J
authors.initials: MJ
authors.lastname: Currie
authors.affiliation: Mackenzie Cancer Research Group, Department of Pathology and Biomedical Science, University of Otago Christchurch , Christchurch , New Zealand ; Canterbury Regional Cancer and Blood Service, Canterbury District Health Board, Christchurch Hospital , Christchurch , New Zealand.
authors.forename: Bridget A
authors.initials: BA
authors.lastname: Robinson
authors.affiliation: Biostatistics and Computational Biology Unit, Department of Public Health and General Practice, University of Otago Christchurch , Christchurch , New Zealand.
authors.forename: John F
authors.initials: JF
authors.lastname: Pearson
authors.affiliation: Centre for Free Radical Research, Department of Pathology and Biomedical Science, University of Otago Christchurch , Christchurch , New Zealand.
authors.forename: Margreet C M
authors.initials: MC
authors.lastname: Vissers
keywords: BNIP3; VEGF; ascorbate; colorectal cancer; hydroxylation; hypoxia-inducible factor