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Abstract

Breast cancer (BC) is a leading cause of cancer-related deaths globally.  Epigenetic alterations, particularly DNA hypermethylation, contribute to BC development by silencing tumor suppressor genes. This study investigated the ability of disulfiram (DSF) to reverse epigenetic silencing of the RASSF1A gene in the MCF-7 breast cancer cell line.  Using MTT assays, DSF was found IC50 at doses of 5, 10, 12.5, and 15µM after 72 hours of treatment.  Methylation-specific PCR (MSP) and real-time PCR revealed partial demethylation of the RASSF1A gene promoter at a DSF dose of 15µM after 24 hours and all doses (5-15µM) after 72 hours.  This demethylation was accompanied by increased RASSF1A gene expression at the highest DSF dose (15µM) after 24 hours and at doses of 10, 12.5, and 15µM after 72 hours. These findings suggest that DSF can effectively induce epigenetic reversion of RASSF1A silencing, restoring its tumor suppressor function.  This data highlights the potential of DSF as a novel therapeutic agent for breast cancer, either as a monotherapy or in combination with other therapies.