Preliminary Evaluation of SGLT2 Inhibitors in Human Bladder Cancer T24 Cells.

Background/Objectives: Sodium-glucose cotransporter 2 (SGLT2) inhibitors have recently gained attention for their potential anticancer properties. However, their effects in bladder cancer remain poorly understood. In this preliminary study, we assessed the effects of canagliflozin, dapagliflozin, and empagliflozin on metabolic activity, cell cycle distributions, migratory behavior, and SGLT2 expression in the human bladder cancer cell line T24. Methods: Cellular metabolic activity, cell cycle distribution, apoptosis and necrosis, migration, and SGLT2 protein expression were assessed using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, wound-healing and single-cell migration assays, Western blotting, and fluorescence microscopy. Bioinformatic analyses of SLC5A2 were additionally performed. Results: All three compounds reduced MTT-derived metabolic activity in a concentration-dependent manner and induced G0/G1 cell-cycle accumulation without increasing apoptosis or necrosis after 24 h. Flozins also impaired collective wound closure and altered single-cell migration parameters, although replicate-level analysis showed no significant differences between individual treatment conditions and control after multiple-comparison correction. SGLT2 protein was detected in T24 cells and decreased following treatment, with compound-dependent differences in magnitude. Conclusions: Flozins affect metabolic activity, cell-cycle progression, wound closure, and migratory behavior in T24 cells. However, these findings do not establish SGLT2-specific mechanisms and were obtained at concentrations exceeding typical clinical plasma exposure. Further target-validation and mechanistic studies are required.