Epigenotoxicity of environmental pollutants evaluated by a combination of DNA methylation inhibition and capillary electrophoresis–laser-induced fluorescence immunoassay
- 497 Downloads
A variety of environmental pollutants may cause abnormal DNA methylation, which further disturb gene expression. In this work, we developed a rapid and sensitive method for the characterization and identification of the epigenotoxicity of environmental pollutants in terms of DNA methylation. The method combines in vitro inhibition reactions of a model DNA methyltransferase (DNMT) with rapid and sensitive capillary electrophoresis–laser-induced fluorescence (CE-LIF) immunoassays. This method was first assessed using two known DNMT inhibitors, (–)-epigallocatechin-3-gallate and RG108, and then applied to epigenotoxic evaluation of four aldehydes and six benzo-1,4-quinones. It was found that all these electrophilic chemicals could inhibit DNMT activity, probably due to their interactions with the active sites of DNMT. Interestingly, benzo-1,4-quinones displayed more inhibitory effects on DNMT activity than aldehydes. Among the tested six benzo-1,4-quinones, halogenated benzo-1,4-quinone showed higher inhibitory activity than non-halogenated p-benzo-1,4-quinone. Owing to its speed and sensitivity, our method will be potentially applicable for fast epigenotoxic screening of environmental pollutants and mechanistic study of environmental epigenetics.
KeywordsEnvironmental pollutants Epigenotoxicity DNA methylation DNA methyltransferase Capillary electrophoresis–laser-induced fluorescence immunoassay
The work was supported by the Grants from the National Basic Research Program of China (2009CB421605 and 2010CB933502) and the National Natural Science Foundation of China (21077129, 20877091, 20890112, 21125523, and 20921063) to Dr. H. Wang.
- 2.Singal R, Ginder GD (1999) DNA methylation. Blood 93(12):4059–4070Google Scholar
- 6.Esteller M, Corn PG, Baylin SB, Herman JG (2001) A gene hypermethylation profile of human cancer. Cancer Res 61(8):3225–3229Google Scholar
- 22.Liu Z, Liu S, Xie Z, Blum W, Perrotti D, Paschka P, Klisovic R, Byrd J, Chan KK, Marcucci G (2007) Characterization of in vitro and in vivo hypomethylating effects of decitabine in acute myeloid leukemia by a rapid, specific and sensitive LC-MS/MS method. Nucleic Acids Res 35(5):e31CrossRefGoogle Scholar
- 24.Darii MV, Cherepanova NA, Subach OM, Kirsanova OV, Raskó T, Slaska-Kiss K, Kiss A, Deville-Bonne D, Reboud-Ravaux M, Gromova ES (2009) Mutational analysis of the CG recognizing DNA methyltransferase SssI: insight into enzyme–DNA interactions. Biochim Biophys Acta 1794(11):1654–1662CrossRefGoogle Scholar
- 29.Fang MZ, Wang Y, Ai N, Hou Z, Sun Y, Lu H, Welsh W, Yang CS (2003) Tea polyphenol (−)-epigallocatechin-3-gallate inhibits DNA methyltransferase and reactivates methylation-silenced genes in cancer cell lines. Cancer Res 63(22):7563–7570Google Scholar
- 32.Voulgaridou GP, Anestopoulos I, Franco R, Panayiotidis MI, Pappa A (2011) DNA damage induced by endogenous aldehydes: current state of knowledge. Mutat Res 711(1–2):13–27Google Scholar