A2780

All tissue samples were obtained from patients treated at The First Affiliated Hospital of Zhengzhou University. Each patient signed an informed consent form, and this study was approved by the First Affiliated Hospital of Zhengzhou University Institutional Review Board. Tissues samples were collected from OC patients with unilateral ovarian invasion and normal contralateral ovary, including 17 cases of high-grade serous carcinoma, 2 cases of mucinous cystadenoma and 1 case of mucinous carcinoma. Human OC cell lines (A2780, SKOV3, OVCAR3) and a normal ovary cell line (IOSE80) were purchased from the Cell Bank of Chinese Academy of Sciences (Shanghai, China). All cell lines were authenticated using the short tandem repeat (STR) profiling test. Then, IOSE80, A2780, OVCAR3 cells were cultured in RPMI-1640 Medium (Biological Industries, USA) supplemented with 10% fetal bovine serum (FBS, Biological Industries, USA), 100 units/ml penicillin, and 100 μg/ml streptomycin (Biological Industries, USA). SKOV3 cell was maintained in McCoy’s 5A Medium (Biological Industries, USA) supplemented with 10% FBS, 100 U/ml penicillin, and 100 μg/ml streptomycin.

The cell lines SKOV3, ES2, and HEK293T were obtained from ATCC (Manassas, VA, USA). The A2780 cell line was acquired from ECACC (Salisbury, UK). A2780 and HEK293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Biological Industries (BI), Israel), ES2 cells were cultured in McCoy’s 5A medium (BI), and SKOV3 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (BI). All media were supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY, USA) and cells were cultured at 37 °C in a 5% CO₂ incubator. All cell lines were authenticated by genetic profiling using polymorphic short tandem repeat loci. The cells were treated with LY294002 (Beyotime, Shanghai, China) or rapamycin (Beyotime) at the indicated concentrations for 24 h, before subsequent analysis.