Biomarkers 2015-02-01

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

Alessio Perotti, Valeria Rossi, Antonio Mutti, Annamaria Buschini

Index: Biomarkers 20(1) , 64-70, (2015)

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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.