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Abstract
Identifying novel epigenetic biomarkers is a promising way to improve the clinical management of patients with breast cancer. Our study aimed to determine the methylation pattern of 25 tumor suppressor genes (TSG) and select the best methylation biomarker associated with clinicopathological features in the cohort of Slovak patients diagnosed with invasive ductal carcinoma (IDC). Overall, 166 formalin-fixed, paraffin-embedded (FFPE) tissues obtained from patients with IDC were included in the study. The methylation status of the promoter regions of 25 TSG was analyzed using semiquantitative methylation-specific MLPA (MS-MLPA). We identified CDH13 as the most frequently methylated gene in our cohort of patients. Further analysis by ddPCR confirmed an increased level of methylation in the promoter region of CDH13. A significant difference in CDH13 methylation levels was observed between IDC molecular subtypes LUM A versus HER2 (P = 0.0116) and HER2 versus TNBC (P = 0.0234). In addition, significantly higher methylation was detected in HER2+ versus HER2- tumors (P = 0.0004) and PR− versus PR+ tumors (P = 0.0421). Our results provide evidence that alteration in CDH13 methylation is associated with clinicopathological features in the cohort of Slovak patients with IDC. In addition, using ddPCR as a methylation-sensitive method represents a promising approach characterized by higher precision and technical simplicity to measure the methylation of target CpGs in CDH13 compared to other conventional methods such as MS-MLPA.
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1 Comenius University in Bratislava, Biomedical Centre Martin, Jessenius Faculty of Medicine in Martin, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000 0001 0940 9708); Comenius University in Bratislava, Department of Pathological Physiology, Jessenius Faculty of Medicine in Martin, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000 0001 0940 9708); Comenius University in Bratislava, Biobank for Cancer and Rare Diseases, Jessenius Faculty of Medicine in Martin, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000 0001 0940 9708)
2 Comenius University in Bratislava, Biomedical Centre Martin, Jessenius Faculty of Medicine in Martin, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000 0001 0940 9708); Comenius University in Bratislava, Department of Medical Biology, Jessenius Faculty of Medicine in Martin, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000 0001 0940 9708)
3 Comenius University in Bratislava, Biomedical Centre Martin, Jessenius Faculty of Medicine in Martin, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000 0001 0940 9708)
4 Comenius University in Bratislava, Biomedical Centre Martin, Jessenius Faculty of Medicine in Martin, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000 0001 0940 9708); Comenius University in Bratislava, Clinic of Obstetrics and Gynecology, Jessenius Faculty of Medicine in Martin and Martin University Hospital, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000000109409708)
5 Comenius University in Bratislava, Department of Molecular Biology and Genomics, Jessenius Faculty of Medicine in Martin, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000 0001 0940 9708)
6 Comenius University in Bratislava, Clinic of Obstetrics and Gynecology, Jessenius Faculty of Medicine in Martin and Martin University Hospital, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000000109409708)
7 Comenius University in Bratislava, Biomedical Centre Martin, Jessenius Faculty of Medicine in Martin, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000 0001 0940 9708); Comenius University in Bratislava, Biobank for Cancer and Rare Diseases, Jessenius Faculty of Medicine in Martin, Martin, Slovakia (GRID:grid.7634.6) (ISNI:0000 0001 0940 9708)