Epigenotoxicology needs simple and fast tools to assess xenobiotic epigenetic load. This work proposes a comet assay modification designed to detect global methylation changes (Methysens 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.

Methy-sens Comet assay and DNMTs transcriptional analysis as a combined approach in epigenotoxicology / Perotti, Alessio; Rossi, Valeria; Mutti, Antonio; Buschini, Annamaria. - In: BIOMARKERS. - ISSN 1354-750X. - 20:1(2015), pp. 64-70. [10.3109/1354750X.2014.992813]

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

PEROTTI, Alessio;ROSSI, Valeria;MUTTI, Antonio;BUSCHINI, Annamaria
2015

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 (Methysens 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.
Methy-sens Comet assay and DNMTs transcriptional analysis as a combined approach in epigenotoxicology / Perotti, Alessio; Rossi, Valeria; Mutti, Antonio; Buschini, Annamaria. - In: BIOMARKERS. - ISSN 1354-750X. - 20:1(2015), pp. 64-70. [10.3109/1354750X.2014.992813]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11381/2796894
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