Probenecid Suppresses Migration, invasion and angiogenesis in hepatocellular carcinoma by modulating purinergic signaling and p38 MAPK pathway

Probenecid (PBN) is a well-established therapeutic agent traditionally used to treat gout and to regulate renal excretion by inhibiting ATP-related membrane transporters. While these properties are well documented, their potential relevance in cancer biology remains largely unexplored. Given the critical role of extracellular ATP and purinergic signaling in tumor progression, we hypothesized that PBN might exert antitumor effects by interfering with this pathway. In this study, we investigated the anticancer activity of PBN in HepG2 human hepatocellular carcinoma cells and in an orthotopic mouse model. PBN enhanced cell adhesion and reduced migration and invasion, effects associated with altered integrin expression, selective inhibition of matrix metalloproteinase (MMP) activity, modulation of epithelial-mesenchymal transition (EMT) markers and reduced activation of the p38 MAPK pathway. In vivo, PBN suppressed tumor growth and reduced circulating vascular endothelial growth factor (VEGF) levels, indicating impaired angiogenesis. Altogether, these findings indicate that PBN may represent a promising candidate for drug repurposing in hepatocellular carcinoma. By modulating purinergic signaling and selectively inhibiting p38 MAPK, PBN appears to limit tumor invasiveness and angiogenesis, supporting its potential relevance for further preclinical and clinical investigation.