Human OC xenografts in immunodeficient mice

The animal studies were carried out in compliance with the protocol that underwent review and approval by the Animal Care and Use Committee of the Fourth Military Medical University, adhering to the guidelines established by the National Institutes of Health (eighth edition, NIH, 2001). During the experiment, eight nude mice (6 - 8 weeks old) were randomly divided into two groups, and each mouse received 200 µL RPMI-1640 medium supplemented with Matrigel (Sigma) containing 1 × 10⁵ ES-2 cells (transduced with control or IGF2BP3-targeting shRNA) in the unilateral flank area subcutaneously. The mice were subjected to measurement of tumor volumes every week and monitored for body weight changes and other side effects. Tumor volume was determined by the formula V = 1/2 × L × W², where L and W separately represent tumor length and width. Mice were euthanized after 30 days to avoid severe discomfort in the control group. Tumor tissues were collected for biochemical or histological assays.