Evaluation of Oxidative DNA Damage Using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay, and the Protective Effects of N-Acetylcysteine Amide on Zearalenone-induced Cytotoxicity in Chang Liver Cells

• Published in: Toxicological research (Seoul) Vol. 29; no. 1; pp. 43 - 52
• Main Authors: Kang, C.G., Gyeongsang National University, Jinju, Republic of Korea, Lee, H.K., Gyeongsang National University, Jinju, Republic of Korea, Yoo, Y.S., Gyeongsang National University, Jinju, Republic of Korea, Hah, D.Y., Gyeongnam National University of Science and Technology, Jinju, Republic of Korea, Kim, C.H., Gyeongnam National University of Science and Technology, Jinju, Republic of Korea, Kim, E.Y., Gyeongsang National University, Jinju, Republic of Korea, Kim, J.S., Gyeongsang National University, Jinju, Republic of Korea
• Format: Journal Article
• Language: English
• Published: Singapore 한국독성학회 01.03.2013; Springer Singapore; The Korean Society of Toxicology
• Subjects: An alkaline single cell gel electrophoresis,Comet assay,DNA damaging,N-Acetylcysteine amide,Chang liver cell; Biomedicine; Pharmacology/Toxicology; TOXICIDAD; TOXICITE; TOXICITY; ZEARALENONA; ZEARALENONE; 예방의학; Zearalenone; Cytotoxicity; N-Acetylcysteine amide(NACA); An alkaline single cell gel electrophoresis(SCGE) Comet assay; Chang liver cell; DNA damaging; N-Acetylcysteine amide (NACA); An alkaline single cell gel electrophoresis (SCGE) Comet assay
• ISSN: 1976-8257; 2234-2753
• DOI: 10.5487/TR.2013.29.1.043

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium that are found in cereals and agricultural products. ZEN has been implicated in mycotoxicosis in farm animals and in humans. The toxic effects of ZEN are well known, but the ability of an alkaline Comet assay to assess ZEN-induced oxidative DNA damage in Chang liver cells has not been established. The first aim of this study was to evaluate the Comet assay for the determination of cytotoxicity and extent of DNA damage induced by ZEN toxin, and the second aim was to investigate the ability of N-acetylcysteine amide (NACA) to protect cells from ZEN-induced toxicity. In the Comet assay, DNA damage was assessed by quantifying the tail extent moment (TEM; arbitrary unit) and tail length (TL; arbitrary unit), which are used as indicators of DNA strand breaks in SCGE. The cytotoxic effects of ZEN in Chang liver cells were mediated by inhibition of cell proliferation and induction of oxidative DNA damage. Increasing the concentration of ZEN increased the extent of DNA damage. The extent of DNA migration, and percentage of cells with tails were significantly increased in a concentration-dependent manner following treatment with ZEN toxin (p 0.05). Treatment with a low concentration of ZEN toxin (25 μM) induced a relatively low level of DNA damage, compared to treatment of cells with a high concentration of ZEN toxin (250 μM). Oxidative DNA damage appeared to be a key determinant of ZEN-induced toxicity in Chang liver cells. Significant reductions in cytolethality and oxidative DNA damage were observed when cells were pretreated with NACA prior to exposure to any concentration of ZEN. Our data suggest that ZEN induces DNA damage in Chang liver cells, and that the antioxidant activity of NACA may contribute to the reduction of ZEN-induced DNA damage and cytotoxicity via elimination of oxidative stress.