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Updated: Aug 9, 2025

Visualizing and Quantifying Endonuclease-Based Site-Specific DNA Damage
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A Cell System-Assisted Strategy for Evaluating the Natural Antioxidant-Induced Double-Stranded DNA Break (DSB) Style

Yuduki Someya1, Sakine Kobayashi1, Kazuya Toriumi1

  • 1Faculty of Science and Technology, Gunma University, Kiryu 376-8515, Japan.

Genes
|February 25, 2023
PubMed

Insights

Natural antioxidants like sulforaphane and quercetin can induce DNA damage, contributing to their antitumor effects. This study developed a novel method to analyze these mechanisms, revealing insights into plant-derived compounds.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Plant-derived natural antioxidants exhibit antitumor properties, but their precise molecular mechanisms remain unclear.
  • In vitro assays for identifying antioxidant targets are expensive, time-consuming, and may not accurately represent in vivo conditions.
  • Understanding the DNA-damaging potential of natural antioxidants is crucial for elucidating their anticancer activity.

Purpose of the Study:

  • To investigate the DNA-damaging effects of specific natural antioxidants with known antitumor properties.
  • To establish an evaluation system for analyzing the cytotoxic mechanisms of natural antioxidants.
  • To differentiate between DNA-damaging and non-DNA-damaging cytotoxic effects of these compounds.

Main Methods:

  • Utilized gene-knockout human Nalm-6 and HeLa cell lines.
  • Pretreated cells with the DNA-dependent protein kinase inhibitor NU7026.
  • Assessed DNA damage induction by natural antioxidants including sulforaphane, resveratrol, quercetin, kaempferol, and genistein.

Main Results:

  • Sulforaphane was found to induce single-strand breaks or DNA strand crosslinks.
  • Quercetin was identified as an inducer of double-strand breaks.
  • Resveratrol exhibited cytotoxicity independent of DNA damage, while kaempferol and genistein induced DNA damage through unelucidated pathways.

Conclusions:

  • The developed evaluation system effectively analyzes the DNA-damaging mechanisms of natural antioxidants.
  • Specific antioxidants like sulforaphane and quercetin exert antitumor effects through distinct types of DNA damage.
  • Further research is needed to fully elucidate the DNA-damage mechanisms of kaempferol and genistein.
Keywords:
DNA double-strand breakDNA single-strand breakantitumor effectnatural antioxidantnon-homologous end-joining

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