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A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
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Ascorbic acid does not reduce the anticancer effect of radiotherapy.

Yoichiro Hosokawa1, Ryo Saga1, Satoru Monzen1

  • 1Department of Radiation Science, Graduate School of Health Sciences, Hirosaki University, Hirosaki, Aomori 036-8564, Japan.

Biomedical Reports
|January 27, 2017
PubMed
Summary

Ascorbic acid (AsA) scavenges hydroxyl radicals generated by X-ray irradiation, potentially reducing anticancer effects. This study found AsA alone induced leukemia cell death, suggesting no added benefit with radiation therapy.

Keywords:
X-ray irradiationascorbic acidcell deathhydroxyl radicalradical scavenger

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Radiation Oncology

Background:

  • Ascorbic acid (AsA) is investigated for its potential to mitigate radiation therapy side effects and enhance antitumor activity.
  • The interaction between AsA's radical scavenging properties and the cytotoxic effects of hydroxyl (OH) radicals generated by X-ray irradiation requires elucidation.

Purpose of the Study:

  • To examine the association between ascorbic acid's scavenged activity and the anticancer effects of X-ray-generated hydroxyl radicals.
  • To evaluate the impact of ascorbic acid on human leukemia HL60 cell survival and DNA fragmentation in conjunction with X-ray irradiation.

Main Methods:

  • Human leukemia HL60 cells were subjected to X-ray irradiation and varying concentrations of ascorbic acid.
  • Cell survival and DNA fragmentation were assessed.
  • Electron spin resonance (ESR) spectroscopy was employed to detect and quantify hydroxyl radicals and their scavenging by AsA.

Main Results:

  • Ascorbic acid concentrations greater than 1 mM led to decreased cell survival and increased DNA fragmentation.
  • X-ray irradiation generated OH radicals, which were effectively scavenged by AsA at concentrations above 75 µM.
  • An intracellular AsA concentration of 75 µM, achieved with 5 mM extracellular treatment, induced significant HL60 cell death independently of irradiation.
  • Combined AsA treatment and irradiation did not enhance the viability reduction compared to irradiation alone.

Conclusions:

  • Ascorbic acid exhibits significant cytotoxic effects on HL60 cells, even without radiation.
  • The radical scavenging activity of AsA can interfere with the generation of cytotoxic OH radicals by X-ray irradiation.
  • The combination of AsA with radiotherapy does not appear to enhance anticancer effects and may even diminish them, warranting careful consideration in clinical applications.