CLOCKSHIFT

Breast cancer risk and epigenetic effects of the rotating night shift work and lifestyle.

WP1

Methylation status of core circadian genes and tumor suppressor genes in cross-sectional study of Polish nurses

Leader: Ph.D. Edyta Reszka

The objective of this work package is to analyze of global methylation and promoter methylation of circadian genes and cell cycle regulatory genes (PER1, PER2, PER3, BMAL1, CLOCK, CRY1, CRY2, NPAS2, TP53, CDKN1A, CDKN2A, RB1, BRCA1, BRCA2). Epigenetic analyses will be performed at the Nofer Institute of Occupational Medicine using blood samples collected from 705 women taking part in e cross-sectional study in Poland (for more information, see "Study population"). The methylation analysis will be performed by fluorescence-based, quantitative methylation-specific real-time PCR assay (qMS-PCR) with two sets of primers and a MethyLight fluorescent probe, after sodium bisulfate conversion. Global DNA methylation in human leukocytes will be determined by means of commercial ELISA assays. Methylation levels of samples will be calculated relative to the methylated control DNA and expressed as a percentage of methylated DNA.