Evaluating the Impact of Febuxostat on 5-Aminolevulinic Acid-Based Photodynamic Therapy in Bladder Cancer Cell Lines

Abstract

Background: Bladder cancer (BC) is a prevalent malignancy worldwide, with significant challenges in recurrence management following conventional treatments. This study investigates the potential of Febuxostat, an established xanthine oxidase inhibitor, to enhance the efficacy of photodynamic therapy (PDT) utilizing 5-aminolevu-linic acid (5-ALA) in bladder cancer cell lines T24 and 5637. Methods: In this in vitro study, we evaluated the effect of 5-ALA, Febuxostat and 5-ALA-Febuxostat combination therapy in T24 and 5637 cell lines as representatives of human bladder cancer. The assessment includes scratch-wound assay, colony formation assay, ROS measurement, flow cytometric analysis of apoptosis and DNA cell cycle, real-time PCR (BAX/BCL2, E-cadherin, N-cadherin, HIF1 alpha and VEGFC genes). Results: Our findings demonstrate that co-administration of Febuxostat significantly increases ROS output compared to ALA treatment alone (p-value <= 0.05), leading to enhanced cytotoxicity and apoptosis. Flow cytometric analyses revealed elevated apoptosis rates in combination treatment groups, and cell cycle assessments indicated a preferential sub-G1 phase arrest associated with the enhanced apoptotic response (P Value = 0.03). Additionally, gene expression profiling showed alterations in key apoptotic markers (BAX/BCL2), with an upregulation of pro-apoptotic genes and a downregulation of anti-apoptotic factors and increase E-Cadherin/N-Cadherin in response to Febuxostat-enhanced PDT (P Value>0.05). Conclusion: These results indicate that Febuxostat effectively potentiates the photodynamic effects of ALA in bladder cancer cells, promising a novel therapeutic strategy that warrants further exploration.