2,4-Dichlorophenol induces global DNA hypermethylation through the increase of S-adenosylmethionine and the upregulation of DNMTs mRNA in the liver of goldfish Carassius auratus

Altered DNA methylation is associated with changes in gene expression, signal transduction and stress response after exposure to a wide range of exogenous compounds, and abnormal methylation is a major toxic effect induced by chemicals such as benzene and phenols. 2,4-Dichlorophenol (2,4-DCP), a derivative of phenol, has been classified as a priority pollutant by the US EPA due to its toxic effects on aquatic organisms. However, the effect of 2,4-DCP on DNA methylation and its potential mechanism in fish are rarely understood. The present study aims to figure out whether 2,4-DCP could impact DNA methylation and explore its potential mechanisms by measuring the global DNA methylation levels, S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) contents, the mRNA expression of DNA methyltransferase1 (DNMT1) and DNA methyltransferase3 (DNMT3) in the liver of goldfish Carassius auratus. DNA methylation levels were analyzed using high performance liquid chromatography (HPLC) and MspI/HpaII ethidium bromide assay, SAM and SAH contents were determined by HPLC, the mRNA expression of DNMT1 and DNMT3 was measured by quantitative-PCR (qPCR). The results showed that 2,4-DCP caused global DNA hypermethylation, elevated the methylation levels of CpG islands, increased the SAM and SAH contents, decreased the SAM/SAH ratio, and upregulated the mRNA expression of DNMT1 and DNMT3, while depletion of SAM with Na2SeO3 and inhibition of DNMTs activity with 5-aza-2′-deoxycytidine (5AdC) impaired 2,4-DCP-induced global DNA hypermethylation, suggesting that the increase of SAM contents and upregulation of the mRNA expression of DNMT1 and DNMT3 may play important roles in 2,4-DCP-induced global DNA hypermethylation process. Our report is the first one to show that short-term 2,4-DCP exposure caused the global DNA hypermethylation via altered SAM level and DNMTs expression in fish.