Sv huc 1

Unless otherwise specified, all reagents were standard laboratory stocks. Human uroepithelial (SV-HUC-1) and bladder cancer (T24) cell lines were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured in Ham's F12 medium and Roswell Park Memorial Institute-1640 media, respectively, supplemented with 10% fetal bovine serum and 2% penicillin/streptomycin, and incubated in a humidified atmosphere of 5% CO₂ at 37°C. Cells at <80% confluence were cultured for 24 h in the presence of IL-2, IFN-α, IFN-γ (final activity concentration, 1,000 U/ml; Prospec-Tany TechnoGene, Ltd., East Brunswick, NJ, USA) or PBS as a control. The cells were harvested for analysis by RT-qPCR and western blotting.

The human BC cell line EJ was purchased from the American Type Culture Collection (Manassas, VA, USA). The human urothelium cell line SV-HUC-1(normal control) was purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China), cultured in DMEM medium (containing 5.5 mmol/L D-glucose, normal glucose; NG), and supplemented with 10% FBS, 1% streptomycin, and penicillin in a humidified incubator with 95% air and 5% CO² at 37°C. EJ cells were seeded in complete medium at 70-80% confluence in six-well plates. The function of ALO on the apoptosis and proliferation of EJ cells was determined. After culturing for 24 h, the medium was replaced with serum-free media. The cells were treated with different ALO concentrations (0, 25, 50, and 100 µmol/L) and incubated in a humidified incubator at 37°C for 12, 24, and 48 h. Then, the expression levels of related proteins and experiment were determined. All experiments were conducted in serum-free conditions and at least repeated thrice.

Human bladder cancer cell lines UMUC3, HT1197, T24, J82 and human bladder epithelial cell line SV-HUC-1 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS, Gibco, USA) and were incubated at 37 °C in a humidified atmosphere with 5% CO₂.