Subcutaneous Xenograft Tumor Model

Nude mice were obtained from Shanghai Gene-Pharma Co. (Shanghai, China) and housed in a standard laboratory with free access to food and water. The study was approved by the Animal Care Committee of Lanzhou University Second Hospital and conducted following the ARRIVE guidelines. For the establishment of the subcutaneous xenograft tumor model, resistant LCSCs were resuspended to a density of 5×10⁵/mL in PBS. Each mouse was subcutaneously injected with 100 μL of cells in the right dorsal area. When the tumor volume reached 150 mm³, the mice received sorafenib (100 mg/kg) or PBS orally once daily for 10 days. At the end of the experiment, mice were euthanized, and tumors were resected for testing gene or protein expression levels.