title: Pharmacological doses of daily ascorbate protect tumors from radiation damage after a single dose of radiation in an intracranial mouse glioma model.
abstract: Pharmacological ascorbate is currently used as an anti-cancer treatment, potentially in combination with radiation therapy, by integrative medicine practitioners. In the acidic, metal-rich tumor environment, ascorbate acts as a pro-oxidant, with a mode of action similar to that of ionizing radiation; both treatments kill cells predominantly by free radical-mediated DNA damage. The brain tumor, glioblastoma multiforme (GBM), is very resistant to radiation; radiosensitizing GBM cells will improve survival of GBM patients. Here, we demonstrate that a single fraction (6 Gy) of radiation combined with a 1 h exposure to ascorbate (5 mM) sensitized murine glioma GL261 cells to radiation in survival and colony-forming assays in vitro. In addition, we report the effect of a single fraction (4.5 Gy) of whole brain radiation combined with daily intraperitoneal injections of ascorbate (1 mg/kg) in an intracranial GL261 glioma mouse model. Tumor-bearing C57BL/6 mice were divided into four groups: one group received a single dose of 4.5 Gy to the brain 8 days after tumor implantation, a second group received daily intraperitoneal injections of ascorbate (day 8-45) after implantation, a third group received both treatments and a fourth control group received no treatment. While radiation delayed tumor progression, intraperitoneal ascorbate alone had no effect on tumor progression. Tumor progression was faster in tumor-bearing mice treated with radiation and daily ascorbate than in those treated with radiation alone. Histological analysis showed less necrosis in tumors treated with both radiation and ascorbate, consistent with a radio-protective effect of ascorbate in vivo. Discrepancies between our in vitro and in vivo results may be explained by differences in the tumor microenvironment, which determines whether ascorbate remains outside the cell, acting as a pro-oxidant, or whether it enters the cells and acts as an anti-oxidant.
authors.affiliation: Malaghan Institute of Medical Research , Wellington , New Zealand.
authors.forename: Carole
authors.initials: C
authors.lastname: Grasso
authors.affiliation: School of Biological Sciences, Victoria University , Wellington , New Zealand.
authors.forename: Marie-Sophie
authors.initials: MS
authors.lastname: Fabre
authors.affiliation: School of Biological Sciences, Victoria University , Wellington , New Zealand.
authors.forename: Sarah V
authors.initials: SV
authors.lastname: Collis
authors.affiliation: School of Biological Sciences, Victoria University , Wellington , New Zealand.
authors.forename: M Leticia
authors.initials: ML
authors.lastname: Castro
authors.affiliation: Malaghan Institute of Medical Research , Wellington , New Zealand ; School of Biological Sciences, Victoria University , Wellington , New Zealand.
authors.forename: Cameron S
authors.initials: CS
authors.lastname: Field
authors.affiliation: Department of Radiation Therapy, University of Otago , Wellington , New Zealand.
authors.forename: Nanette
authors.initials: N
authors.lastname: Schleich
authors.affiliation: Malaghan Institute of Medical Research , Wellington , New Zealand ; School of Biological Sciences, Victoria University , Wellington , New Zealand.
authors.forename: Melanie J
authors.initials: MJ
authors.lastname: McConnell
authors.affiliation: Malaghan Institute of Medical Research , Wellington , New Zealand ; Department of Radiation Therapy, University of Otago , Wellington , New Zealand.
authors.forename: Patries M
authors.initials: PM
authors.lastname: Herst
keywords: GL261; intracranial mouse glioma model; pharmacological ascorbate; radiation; radioprotection; radiosensitization