title: DNA damage evaluated by gammaH2AX foci formation by a selective group of chemical/physical stressors.
abstract: It has been reported that the phosphorylated form of histone variant H2AX (gammaH2AX) plays an important role in the recruitment of DNA repair and checkpoint proteins to sites of DNA damage, particularly at double strand breaks (DSBs). Using gammaH2AX foci formation as an indicator for DNA damage, several chemicals/stress factors were chosen to assess their ability to induce gammaH2AX foci in a 24h time frame in a human amnion FL cell line. Two direct-acting genotoxins, methyl methanesulfonate (MMS) and N-ethyl-N-nitrosourea (ENU), can induce gammaH2AX foci formation in a time- and dose-dependent manner. Similarly, an indirect-acting genotoxin, benzo[a]pyrene (BP), also induced the formation of gammaH2AX foci in a time- and dose-dependent manner. Another indirect genotoxin, 2-acetyl-aminofluorene (AAF), did not induce gammaH2AX foci formation in FL cells; however, AAF can induce gammaH2AX foci formation in Chinese hamster CHL cells. Neutral comet assays also revealed the induction of DNA strand breaks by these agents. In contrast, epigenetic carcinogens azathioprine and cyclosporine A, as well as non-carcinogen dimethyl sulfoxide, did not induce gammaH2AX foci formation in FL cells. In addition, heat shock and hypertonic saline did not induce gammaH2AX foci. Cell survival analyses indicated that the induction of gammaH2AX is not correlated with the cytotoxic effects of these agents/factors. Taken together, these results suggest that gammaH2AX foci formation could be used for evaluating DNA damage; however, the different cell types used may play an important role in determining gammaH2AX foci formation induced by a specific agent.
authors.affiliation: Department of Public Health, Institute for Toxicology, Zhejiang University School of Medicine, 353 Yan An Road, Hangzhou, Zhejiang 310031, China.
authors.forename: Chunxian
authors.initials: C
authors.lastname: Zhou
authors.forename: Zhongxiang
authors.initials: Z
authors.lastname: Li
authors.forename: Huiling
authors.initials: H
authors.lastname: Diao
authors.forename: Yanke
authors.initials: Y
authors.lastname: Yu
authors.forename: Wen
authors.initials: W
authors.lastname: Zhu
authors.forename: Yayun
authors.initials: Y
authors.lastname: Dai
authors.forename: Fanqing F
authors.initials: FF
authors.lastname: Chen
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
mesh_terms: D015073:2-Acetylaminofluorene / Q000633:toxicity; D000650:Amnion / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D000906:Antibodies / Q000494:pharmacology; D002273:Carcinogens / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004249:DNA Damage; D005038:Ethylnitrosourea / Q000633:toxicity; D005819:Genetic Markers; D006657:Histones / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009153:Mutagens / Q000633:toxicity
chemical_list: D000906:Antibodies; D002273:Carcinogens; D005819:Genetic Markers; C468783:H2AX protein, human; D006657:Histones; D009153:Mutagens; D015073:2-Acetylaminofluorene; D005038:Ethylnitrosourea