The role of oxidative stress in deoxynivalenol-induced DNA damage in HepG2 cells.
Deoxynivalenol (DON) is a trichothecene mycotoxin and a cereals contamination, whose cytotoxicity has been shown in animals and various cells. However, with respect to the deoxynivalenol-induced DNA damage, especially in humans, are not well understood. The aim of this study was to assess the role of oxidative stress in deoxynivalenol-induced DNA damage, using human hepatoma HepG2 cells. Exposure of the cells to DON caused significant increase of DNA migration in comet assay at concentrations of 3.75-30 microM, which suggests that DON caused DNA strand breaks. To elucidate the role of antioxidation in those effects, DNA migration was monitored by pre-treatment with hydroxytyrosol (HT) as an antioxidant in comet assay. It was found that DNA migration with pre-treatment of HT was dramatically decreased. The DNA damage induced by DON was almost completely prevented. In order to clarify the underlying mechanisms, we evaluated the level of reactive oxygen species (ROS) production with the 2,7-dichlorofluorescein diacetate (DCFH-DA) assay. Significant increase in the level of ROS was observed in HepG2 cells at a higher concentration (60 microM). The involvement of lipid peroxidation in the DNA damage of DON was confirmed by using immunoperoxidase staining for 8-hydroxydeoxyguanosine (8-OHdG) and by measuring levels of thiobarbituric acid-reactive substances (TBARS), the doses being 7.5-60 microM and 3.75-15 microM, respectively. These results indicate that the DNA damage induced by DON in HepG2 cells is probably related to the oxidative stress.
Zhang
Xiaoou
X
Department of Toxicology, Dalian Medical University, No. 9, Dalian 116044, Liaoning, China.
Jiang
Liping
L
Geng
Chengyan
C
Cao
Jun
J
Zhong
Laifu
L
D002460:Cell Line; D020552:Comet Assay; D004249:DNA Damage; D005670:Fusarium; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D009183:Mycotoxins / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism; D014255:Trichothecenes / Q000494:pharmacology*
D009183:Mycotoxins; D017382:Reactive Oxygen Species; D014255:Trichothecenes; C007262:deoxynivalenol