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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, but not dehydroascorbic acid increases intracellular vitamin C content to decrease Hypoxia Inducible

Adam P Fischer1, Sarah L Miles1

  • 1Department of Biomedical Sciences, Joan C. Edwards School of Medicine, Marshall University, One John Marshall Drive, Huntington, WV 25755, United States.

Biomedicine & Pharmacotherapy = Biomedecine & Pharmacotherapie
|December 26, 2016
PubMed
Summary
This summary is machine-generated.

Ascorbic acid (AA) effectively reduces melanoma's malignant potential by inhibiting HIF-1α activity, unlike dehydroascorbic acid (DHA). This highlights AA's promise in cancer therapy by restoring vitamin C regulation.

Keywords:
Ascorbic acidDehydroascorbic acidHypoxia inducible factor-1 alphaMelanomaSVCT2

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Area of Science:

  • Oncology
  • Biochemistry
  • Cancer Research

Background:

  • Hypoxia-inducible factor-1 alpha (HIF-1α) accumulation in tumors promotes cancer progression and reduces patient survival.
  • Ascorbic acid (AA) depletion may drive aberrant HIF-1α activity under normal oxygen conditions.
  • While AA reduces HIF-1α in melanoma, dehydroascorbic acid (DHA)'s therapeutic potential is unevaluated.

Purpose of the Study:

  • To compare the efficacy of AA and DHA in increasing intracellular vitamin C levels.
  • To assess their ability to reduce human melanoma's malignant potential by modulating HIF-1α activity.

Main Methods:

  • Western blot and reporter gene assays were used to evaluate HIF-1α.
  • Vitamin C transporters (SVCT, GLUT) were analyzed via western blot and confocal imaging.
  • Intracellular vitamin C content, HIF-1α activity, and malignant potential (3D spheroid invasion) were measured post-supplementation.

Main Results:

  • Melanoma cells transport both AA and DHA, but only AA favorably impacted advanced melanomas.
  • Physiological glucose levels hindered DHA's ability to increase intracellular vitamin C.
  • AA and ascorbate 2-phosphate (A2P), but not DHA, reduced HIF-1α activity and melanoma's malignant potential.

Conclusions:

  • AA and A2P modulate HIF-1α activity and reduce melanoma's malignant potential.
  • DHA showed limited efficacy due to impaired intracellular vitamin C accumulation.
  • Restoring AA-dependent HIF-1α regulation may improve cancer therapy outcomes and reduce chemoresistance.