Anti-Tumor Effects of CTSL Inhibitor and Adriamycin in Neuroblastoma

All mice experiments were conducted in accordance with the humane treatment of animals under institutional guidelines approved by the Ethical Committee of Children’s Hospital of Soochow University. The mice were housed in individually ventilated cages in the Animal Laboratory of the Children’s Hospital of Soochow University. Six-week-old male M-NSG (NOD-Prkdc^scidIl2rg^em1/Smoc) mice (Shanghai Model Organisms, Shanghai) were used in the study. Subcutaneous tumor transplantation was conducted using the SK-N-BE (2) cells. Cells (n = 1 × 10⁷) were resuspended in 100 μl PBS and implanted into the right flank of nude mice under sterile conditions. After the formation of palpable tumors (tumor volume reached 100 mm³), mice were randomized into four groups (5 mice per group): control group (saline, i.p.), Z-FY-CHO (a specific CTSL inhibitor, HY-128140, MedChemExpress, Shanghai) group (5 mg/kg, i.p.), ADM group (1 mg/kg, i.p.), Z-FY-CHO (5 mg/kg, i.p.) plus ADM (1 mg/kg, i.p.) group. Mice were injected with vehicle or with drugs three times weekly. The size of the tumor and the body weight of each mouse were measured as described previously (Wang et al., 2019 (link)). Mice were sacrificed on day 15, and tumor tissues were harvested.