2.12. In vivo xenograft tumor model

BALB/c nude mice (female, 4 weeks old) were purchased from the Institute of Zoology, Zhejiang Chinese Medical University. A suspension containing 5 × 10⁶ HeLa or ANXA6 knockdown HeLa cells was subcutaneously (s.c.) injected into the right flanks of the nude mice to construct two different types of tumor models, respectively. After 1 week, tumor‐bearing mice were randomly divided into two groups: PBS treatment and rapamycin treatment (5 mg/kg/per 2 days). A vernier caliper was used to measure the tumor dimensions twice per week. After 30 days tumor inoculation, mice were sacrificed and tumors were excised and weighed. According to the criteria outlined in the “Guide for the Care and Use of Laboratory Animals,” all animals received humane care. All experiments were conducted following the official recommendations of the Chinese Zoological Society.