Autoxidation of gallic acid induces ROS-dependent death in human prostate cancer LNCaP cells

Anticancer Res. 2012 May;32(5):1595-602.

Abstract

Background: Prostate cancer is the second most common cause of mortality. Gallic acid (GA) is a natural polyphenol, and we tested its in-vitro cytotoxicity after 24 h in prostate cancer LNCaP cells.

Materials and methods: GA autoxidation was measured fluorimetrically for H(2)O(2), and O(2)(•-) radicals by chemiluminescence. Intracellular reactive oxygen species (ROS) levels were detected with 2',7'-dichlorodihydrofluorescein diacetate. Cytotoxicity was evaluated by crystal-violet, while apoptosis and mitochondrial membrane potential were determined by flow cytometry. Cytochrome c release was detected by enzyme-linked immunosorbent assay, and caspase-8, -9 and -3 activities were measured calorimetrically.

Results: GA autoxidation produced significant levels of H(2)O(2) and O2.-. Increased intracellular ROS levels with GA were reduced by N-acetyl-L-cysteine (NAC) and L-glutathione (GSH). Cells were protected against GA cytotoxicity when pretreated with increasing levels of superoxide dismutase/catalase mixture, NAC, or GSH for 3 h. The number of apoptotic cells increased with GA dose. GA caused mitochondrial potential loss, cytochrome c release, and activation of caspases 3, 8 and 9.

Conclusion: The ROS-dependent apoptotic mechanism of GA kills malignant cells effectively; it is likely that GA could be a good anticancer agent.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acetylcysteine / pharmacology
  • Apoptosis / drug effects*
  • Caspases / metabolism
  • Cell Line, Tumor
  • Cytochromes c / metabolism
  • Gallic Acid / chemistry
  • Gallic Acid / pharmacology*
  • Glutathione / pharmacology
  • Humans
  • Male
  • Membrane Potential, Mitochondrial / drug effects
  • Oxidation-Reduction
  • Prostatic Neoplasms / drug therapy*
  • Prostatic Neoplasms / pathology
  • Reactive Oxygen Species / metabolism*

Substances

  • Reactive Oxygen Species
  • Gallic Acid
  • Cytochromes c
  • Caspases
  • Glutathione
  • Acetylcysteine