Reducing the Effective Dosage of Mitomycin C on a High-Grade Bladder Cancer Cell Line Through Combination With Selenium Nanoparticles: An in Vitro Study Publisher Pubmed



Oskouie IM^{1, 2} ; Amirzargar H² ; Dezfuli AS² ; Mashhadi R^{1, 2} ; Mirzaei A^{1, 2} ; Shamshirgaran A^{1, 2} ; Nikoofar P^{2, 3, 4} ; Aghamir SMK^{1, 2, 5} Show Affiliations
Source: Medical Oncology Published:2025

Abstract

This study aimed to assess the effectiveness of combining selenium nanoparticles (SeNPs) with mitomycin C (MMC) in treating the T24 high-grade bladder cancer cell line to decrease MMC dosage and alleviate its side effects. The T24 (EJ138) cell line was exposed to various concentrations of SeNPs and MMC to identify the IC50 values via the MTT assay. The IC50 of MMC was then lowered by 25%, 50%, and 75%, and different SeNPs concentrations were added, to find the new IC50 values of these combinations. Apoptosis rates were measured using Annexin-V/PI staining, while the DNA cell cycle was analyzed using the PI staining method. The scratch-wound assay, colony-forming assay, and Hoechst staining were employed to examine the cell migration, proliferative capacity, and nuclear morphology, respectively. Real-time PCR assessed the expression levels of SNAIL, E-cadherin, and genes related to angiogenesis and proliferation (VEGF-C and HIF-1α), alongside the apoptosis markers (Bcl-2 and BAX). The co-administration of SeNPs and MMC (178.8 µM SeNPs + 14.9 µM MMC) significantly increased the rate of early apoptosis in the T24 cell line compared to MMC alone (29.8 µM, p < 0.0001). Additionally, SeNPs and MMC induced cell cycle arrest at the SubG1/G1 and G2/M phases, respectively. This effect was observed in the combination group at both phases. Similar to MMC alone, the combination group inhibited cell proliferation, colony formation, and migration in T24 cells (p > 0.05). Our findings indicate that the treatment with the combination increased the expression of apoptosis-related genes and decreased angiogenesis and proliferation-related gene expression similar to MMC alone (p > 0.05). The combined administration of MMC and SeNPs enhances the antitumor efficacy on the T24 cell line. It is proposed that the concurrent use of SeNPs and MMC could effectively reduce the required dosage of MMC, thus minimizing its negative side effects. © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2025.