Methy-sens Comet assay and DNMTs transcriptional analysis as a combined approach in epigenotoxicology

Abstract

Epigenotoxicology needs simple and fast tools to assess xenobiotic epigenetic load. This work proposes a comet assay modification designed to detect global methylation changes (Methy-sens Comet) through enzymatic digestion with two restriction enzymes (HpaII, MspI). In the methylation-sensitive protocol tested for repeatability on A549 cells, nickel chloride induced hypermethylation and decitabine-induced hypomethylation. A concomitant assessment of DNA methyltransferases (DNMTs) genes transcriptional levels has been performed, to implement a multifunctional approach to epigenotoxicology. Methy-sens Comet showed a general good repeatability and sensitivity to methylation changes while DNMTs transcriptional levels granted additional proof of xenobiotic-induced impairment of methylome maintenance.

• Comet assay
• DNMT
• epigenotoxicology
• methylation

Acknowledgements

We would like to thank Dr Mirca Lazzaretti and Mrs Antonietta Cirasolo for technical support.

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article.