Effect of some metabolic inhibitors on apoptosis in breast cancer cell line

Cancer, particularly breast cancer, is a global health concern, causing millions of deaths annually and significantly impacting societal productivity. Treatment options such as single or multiple therapies are available depending on the type, stage, and location of the cancer. Treatments like surgery are used for non-metastatic breast cancers. Metastasis is responsible for more than 90% of cancer-related deaths. Chemotherapy is used for treating metastatic cancer because it can reach metastatic areas. Anticancer drugs promote apoptosis by reactivating apoptotic mechanisms inhibited during oncogenesis. Metabolic pathways can be targeted for effective cancer treatment by inhibiting cell growth and proliferation. Metabolic inhibitors like 2-deoxy-D-glucose and Lonidamine inhibit essential enzymes in metabolic pathways, reducing the availability of downstream products necessary for tumor growth. This study investigated the roles of 2-deoxy-D-glucose and Lonidamine inhibitors in apoptosis in the MCF-7 cell line. The cytotoxic effects of Lonidamine and 2-deoxy-D-glucose inhibitors on MCF-7 cells were assessed using the MTT cytotoxicity assay and xCELLigence real-time cell analyzer (RTCA) methods. From the RTCA analysis, 2-deoxy-D-glucose and Lonidamine were cytotoxic to MCF-7 cancer cells, with 24-hour IC50 values of 5.9 mM and 142.3 µM, respectively. The study utilized Annexin-V FITC labeling followed by flow cytometry analysis to evaluate cell death through apoptosis, revealing that 2-deoxy-D-glucose and Lonidamine cell death occurs through apoptosis. The expressions of caspase-3 and caspase-9 were assessed. Evidenced by the mitochondrial membrane potential reduction, expression of caspase-9, and subsequent activity of caspase-3, our results indicated that 2-deoxy-D-glucose and Lonidamine caused cell death by activating apoptosis intrinsic pathways.