Tumor xenograft model and treatment

All animal experiments followed the guidelines of the Institutional Animal Care and Use Committee of Ren Ji Hospital, and all animal protocols were approved by the same committee. 4–6-week-old nude female mice were ordered from the Chinese Academy of Sciences. Each mouse was injected intraperitoneally with 3.5×10⁶ SKOV3 cells in 200 μL of PBS to establish intraperitoneal xenograft models. After 7 days, the mice were randomly divided into two groups: control group and ABC294640 treatment group (n=6 in each group). ABC294640 was dissolved as a solution with a concentration of 5 mg/mL as follows: 2% DMSO+30% PEG 300+5% Tween 80+PBS. The mice in the control group were injected intraperitoneally with an equivalent amount of DMSO in PBS (vehicle). The mice in the ABC294640 treatment group were treated with ABC294640 intraperitoneally at the dose of 50 mg/kg, 3 times per week for 4 weeks. The mice were sacrificed 28 days after administering ABC294640 (35 days after injecting cancer cells). The difference in the number of visible tumors and the weight between the two groups was calculated. The maximum tumor in each mouse was immersed in tumor lysate and subjected to Western blot analysis, the rest was fixed in formalin and embedded in paraffin. The protein expression levels in tumors of the two groups were also detected through Western blot analysis and immunohistochemistry.