Tuesday, December 15, 2020

DNA methylation patterns of RAR‐β2 and RASSF1A gene promoters in FNAB samples from Greek population with benign or malignant breast lesions

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Abstract

Background

Promoter hypermethylation is common in Breast Cancer (BC) with studies mainly in histological specimens showing frequent methylation of tumor suppressor genes (TSGs) compared with normal tissues. The aim of this study was to estimate the frequency of promoter methylation of RAR‐β2 and RASSF1A genes in breast FNAB material aiming to evaluate the methylation status of these two genes as biomarker for detecting BC in Greek population.

Methods

FNAB material from 104 patients was collected for cytological evaluation and epigenetic analysis. DNA was extracted and subjected to bisulfite conversion. A methylation‐specific PCR was carried out and the final products were separated with electrophoresis in 2% agarose gels.

Results

From 104 samples, RASSF1A hypermethylation was observed in 78 (75%) and RAR‐β2 hypermethylation in 64 (61.6%). 84% and 78% of the cases diagnosed with breast malignancy (n = 50) were methylated for RASSF1A and RAR‐β2, respectively. Methylated RASSF1A and RAR‐β2 were also detected in 88.3% and 76.5% in samples diagnosed as suspicious for malignancy (n = 17) and in 57.2% of samples diagnosed with atypia (n = 14). The Odds Ratio for breast malignancy was 4.545 in patients with RASSF1A hypermethylation and 9.167 in patients with RAR‐β2 hypermethylation underlying their promoter's methylation positive correlation with breast malignancy.

Conclusion

To optimize the sensitivity and specificity of this epigenetic setting, more TSGs related to BC should be gradually imported in our evaluated methylation panel and be validated in a larger study sample with the aim that the obtained epigenetic profiles will provide clinicians with valuable tools for management of BC patients in Greece.

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