Investigating the anticancer effects of 3-Bromopyruvic acid on metabolic alterations in pancreatic cancer cells in vitro

Abstract

Objective: Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive cancers, characterised by a high mortality rate and resistance to chemotherapeutics. Frequent mutations in TP53, SMAD4, and CDKN2A drive metabolic reprogramming and contribute to tumour progression and treatment resistance. Hexokinase 2 (HK2), a key glycolytic enzyme, is commonly overexpressed in PDAC and represents a promising therapeutic target. This study aimed to investigate the anticancer effects of 3-bro-mopyruvic acid (3-BrPA) on metabolic pathways, oxidative stress, and ferroptosis in BXPC-3 PDAC cells harbouring TP53, SMAD4, and CDKN2A mutations. Material and Methods: BXPC-3 cells were administered with various concentrations of3-BrPA, and cell viability was assessed using the MTT assay to determine the IC50. The morphological changes were evaluated via light microscopy. HK2 expression and the enzymatic activities of GST, G6PD, 6-PGD, and GR were measured, along with the glutathione redox status and intracellular iron levels. CPT1C activity was additionally assessed to evaluate the metabolic alterations associated with the fatty acid oxidation pathways. Results: 3-BrPA administration significantly decreased cell viability in a dose-dependent manner and downregulated HK2 expression. It inhibited glycolysis and the pentose phosphate pathway enzymes, reduced the antioxidant capacity, and elevated oxidative stress markers. These changes were associated with increased intracellular iron levels and decreased GSH/GSSG ratios, indicating the induction of ferroptosis. Furthermore, CPT1C activity was upregulated, indicating increased fatty acid oxidation. Conclusion: Our findings demonstrate that 3-BrPA disrupts glu-cose metabolism, induces oxidative stress, and triggers ferroptosis in PDAC cells through HK2 inhibition. This study highlights 3-BrPA's therapeutic potential in targeting cancer metabolism and ferrop-tosis.