CD39 inhibitor (POM-1) modulates purinergic signaling and alters cellular function in murine B16-F10 melanoma cells

Purinergic signaling plays a key role in cellular processes including proliferation, migration, and survival, which are essential for tumor progression. The aim of this study was to investigate effects of polyoxotungstate-1 (POM-1), a purinergic enzyme inhibitor, on viability, migration, and enzymatic activity of B16-F10 melanoma cells. To simulate a tumor microenvironment with impaired extracellular nucleotide degradation and ATP accumulation, effects of POM-1 (50-1000 µM), ATP (1-1000 µM), and their combined treatment were determined after 24 and 72 h. POM-1 effects on purinergic enzymes, oxidative stress, cytokine secretion, and tumor-related cellular functions were measured. POM-1 reduced cell viability, particularly at 250 and 1000 µM after 72 h, with reductions reaching 79% at 1000 µM. ATP exhibited a biphasic effect, with low concentrations enhancing cell viability and function, while high concentrations induced cytotoxicity, diminishing viability by 60% at 1000 µM after 72 h. Combined treatment with POM-1 and ATP enhanced cytotoxicity, reaching approximately 90-92% inhibition cell proliferation. POM-1 elevated ROS and NO levels by 113% and 202%, respectively, and enhanced LDH activity release by up to 144%, indicating cellular damage, and impaired migration, adhesion, and clonogenic capacity, including up to 89% inhibition cell migration and 83% reduction in colony formation. Enzymatic analyses demonstrated that POM-1 inhibited NTPDase activity, altering extracellular nucleotide dynamics, reaching approximately 95% decreased ATP hydrolysis and 80% diminished AMP hydrolysis at higher concentrations. These findings demonstrate that exposure to POM-1 modulates purinergic signaling and induces cytotoxic and functional alterations in murine B16-F10 melanoma cells.