DNA damage evaluated by γH2AX foci formation by a selective group of chemical/physical stressors

作者：Chunxian Zhou; Zhongxiang Li; Huiling Diao; Yanke Yu; Wen Zhu; Yayun Dai; Fanqing F. Chen; Jun Yang

来源：Mutation Research: Genetic Toxicology and Environmental Mutagenesis , 2006, 604(1–2): 8-18.

DOI：10.1016/j.mrgentox.2005.12.004

• γH2AX
• Genotoxin
• Double strand breaks
• Cytotoxicity

It has been reported that the phosphorylated form of histone variant H2AX (γH2AX) 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 γH2AX foci formation as an indicator for DNA damage, several chemicals/stress factors were chosen to assess their ability to induce γH2AX foci in a 24 h time frame in a human amnion FL cell line. Two direct-acting genotoxins, methyl methanesulfonate (MMS) and N-ethyl-N-nitrosourea (ENU), can induce γH2AX foci formation in a time- and dose-dependent manner. Similarly, an indirect-acting genotoxin, benzo[a]pyrene (BP), also induced the formation of γH2AX foci in a time- and dose-dependent manner. Another indirect genotoxin, 2-acetyl-aminofluorene (AAF), did not induce γH2AX foci formation in FL cells; however, AAF can induce γH2AX 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 γH2AX foci formation in FL cells. In addition, heat shock and hypertonic saline did not induce γH2AX foci. Cell survival analyses indicated that the induction of γH2AX is not correlated with the cytotoxic effects of these agents/factors. Taken together, these results suggest that γH2AX foci formation could be used for evaluating DNA damage; however, the different cell types used may play an important role in determining γH2AX foci formation induced by a specific agent.