QUANTIFICATION OF DNA DOUBLE-STRAND BREAKS IN BENIGN AND MALIGNANT BREAST DISEASES

Abstract

Relevance: Double-strand DNA breaks are the most dangerous DNA damage. Analysis of foci of phosphorylated histone protein H2AX (γH2AX) is currently the most sensitive method for detecting DNA double-strand breaks. This protein modification can become a biomarker of cellular stress, especially in diagnosing and monitoring neoplastic diseases. In this study, we utilized novel pattern recognition algorithms on the AKLIDES® platform to automatically analyze immunofluorescent images of γH2AX foci and compare the results with visual scores. The γH2AX foci formation on peripheral blood mononuclear cells of women with breast cancer or benign breast tumors was studied. The article aimed to quantify DNA double-strand breaks in peripheral blood lymphocytes in women with breast cancer and benign breast masses to identify a possible biomarker. Methods: γ-H2AX foci in lymphocytes were analyzed using the automated AKLIDES system in patients with breast cancer (n=29) and benign breast tumors (n=24). Results: When comparing the indicators of the main and control groups in the channel of ruptures “FITC,” a statistically significant difference was found in the indicators “Foci dia” (p=0.0382), “Focilnt mean” (p=0.0166), “Colocalisation” (p=0.0486). In the repair channel “AРС,” significant differences were found in the indicators “Nuclei BGInt” (p=0.0166) and the indicator “Focilnt mean” (p=0.0118). Conclusion: The revealed changes of DNA double-strand breaks along the FITC break channels and APC repair between the main and control groups can possibly serve as a breast cancer diagnostic marker.