Xenograft tumor model

Animal experiments were carried out according to a protocol approved by the Institutional Animal Care and Use Committee of Hefei Institutes of Physical Science. Briefly, female nu/nu athymic mice at the age of 4–6 weeks were purchased from Vital River (Shanghai, China) and maintained in the research animal facility of Hefei Institutes of Physical Science, Chinese Academy of Sciences, under standard environmental conditions. A nude mice model with xenograft tumor was established by the subcutaneous inoculation of 2 × 10⁶ DMS273 cells. The tumor-bearing mice were randomly assigned to four groups (Vehicle, ZIO-101, BMN673, ZIO-101 plus BMN673 combination group). Before administration, ZIO-101 and BMN673 were dissolved in PBS, then administered via intraperitoneal injection at a dose of 50 and 330 mg/kg every other day, respectively. The Vehicle group received the same volume of PBS, and the combination group received the same dose. Tumor sizes were measured using a caliper every other day, and tumor volumes were calculated using the following equation: tumor volume [mm³] = (tumor length × tumor width²) / 2.