Tumor xenograft experiment

BALB/c athymic male nude mice (4–5 weeks old, 20 g) from Huai’an People’s Hospital of Hongze District were used to construct xenograft mouse models. These animals were raised in a specific-pathogen-free (SPF) environment. HCT116 cells (1 × 10⁶) with overexpressed HMX3 were injected subcutaneously into the flanks of nude mice. The tumor size was measured using a vernier caliper every 4 days. The xenograft volumes were calculated using the formula (L× W²)/2. The mice were euthanized by cervical dislocation after feeding for 28 days, and the xenografts were dissected and weighed. All experiments were approved by the Animal Care and Use Committee of Huai’an People’s Hospital of Hongze District. Animal welfare and experimental procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals.