RESUMO
PURPOSE: Aberrant DNA methylation in promoter regions has been found in many cancers, including breast cancer (BC). A Methylation Specific PCR (MSP) was applied in breast Fine Needle Aspiration Biopsy (FNAB) material, which has been rarely used in the literature, to estimate the methylation frequencies of CND2, APC, HIN1 & CDH13 and to assess whether this multiplex methylation panel can be possibly used as an indicator-biomarker for BC detection in a Greek population. METHODS: A total of 104 participants were subjected to FNAB and both cytological evaluation and epigenetic analysis were carried out. DNA was extracted from FNAB samples and was subjected to bisulfite conversion. MSP was carried out with primers specific for either the methylated or unmethylated status for each gene. The final MSP products were analyzed in 2% agarose gels with electrophoresis. RESULTS: Hypermethylation was observed in 74%, 69.2%, 59.6% and 63.4% of the samples for CND2, HIN1, APC and CDH13, respectively. CND2 was the most hypermethylated in C5 cases (90%) and APC and HIN1 in C4 cases (88.2%). A significant correlation between histologic evaluation and the methylation frequencies for all 4 genes was calculated (p<0.001). Odds ratio for breast malignancy was 8.267 for CND2, 5.235 for APC, 7.852 for HIN1 and 22.920 for CDH13, underlying that their methylation is positively related to breast malignancy. Also, it seems that the combination of all genes into a multiplex methylation panel has significantly higher SP and PPV than any single gene methylation. CONCLUSIONS: Our study shows that breast FNAB combined with methylation data from the collected aspirates has a promising potential as a biomarker for the early detection of BC risk in women with suspicious lesions.
