Investigation of ferroptosis cell death pathway in multiple myeloma cells treated with disulfiram

Multiple myeloma (MM), the second most common type of blood cancer in the world, is caused by the proliferation of abnormal plasma cells in the bone marrow. Altho-ugh recent therapeutic advances in the treatment of MM have been highly effective in inc-reasing the survival of patients with MM, MM remains incurable, as most patients ine-vitably relapse and become resistant to current therapies. Therefore, there is a need to deve-lop new agents that target different molecular pathways that drive malignancy in MM. Disulfiram (DSF) has been widely used for years in the US Food and Drug Admi-nistration (FDA) approved treatment of alcoholism and is known to exhibit anticancer ef-fects in various types of cancer, including MM. Recently, it has been shown that DSF suppresses cancer development and metastasis by stimulating ferroptosis, especially in can-cer cells. Ferroptosis is a regulated non-apoptotic form of cell death characterized by increa-sed iron-dependent lipid peroxidation. Since the effect of DSF on ferroptosis in MM cells is unknown, the aim of this thesis is to investigate the effect of DSF on ferroptosis cell de-ath in MM. After the anticancer activity of DSF in MM cell lines (NCI H929, U266 and RPMI 8226) was confirmed by the MTT method, the determination of the amount of malondial-dehyde (MDA) used as a lipid peroxidation marker, H2DFCDA, which is a cytosolic ROS marker, and JC1, which is an indicator of mitochondrial membrane density, were evaluated in Flow Cytometry. Finally, mRNA expression levels of ferroptotic biomarkers were de-termined by quantitative simultaneous PCR. The results showed that DSF significantly increased lipid peroxidation, cytosolic ROS, and mitochondrial membrane density in all three MM cell lines, and decreased the mRNA expression levels of ferroptotic biomarkers GPX4, transferrin SAT1, and SLC7A11. In conclusion, our data show that DSF stimulates ferroptosis in MM cells.