Nude mouse xenograft model

The animal studies were conducted according to the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health according to the procedures approved by the Research Ethical Committee of Nanjing Medical University. The 5- to 6-week-old female SCID nude mice (n=104) were purchased from the Model Animal Institute of Nanjing University, and fed with either a normal or high-fat diet (HFD) as required by the model. SKOV3 cells (7×10⁶) alone or SKOV3 cells (7×10⁶) combined with the well-differentiated HAds (wild-type or gene knock-down type; 7×10⁶) were suspended in 200 µl Matrigel (BD Biosciences), and injected into the subcutaneous tissues in the right lower limbs of the mice. The subcutaneous xenograft growth was monitored using a vernier caliper, three times a week. The tumor volume was calculated according to the following formula: 0.5×LxW² (where L is the tumor dimension at the longest point, and W is the tumor dimension at the widest point). Mice were randomly grouped for medication when the xenograft had grown to an average volume of 100 mm³. Following which, the model groups were given 0.9% normal saline, intraperitoneal injection of carboplatin (Aladdin) once a week (50 mg/kg), as well as an injection of anti-ANGPTL4 neutralizing antibody (BioXcell) and C-terminus ANGPTL4 (cANGPTL4; GenScript) into each tumor at 15 µg/ml before the application of chemotherapy. The animals would be deemed as dead when the xenograft volume was >2,000 mm³. Subsequently, animals were sacrificed using CO₂ suffocation (the flow rate of CO₂, 30% of the cage volume/min), and the xenografts were obtained and preserved in liquid nitrogen for subsequent molecular biological experiments.