Ovcar 3

Five human cell lines containing OC (C200, Caov3, SKOV3, HO8910, and OVCAR3) were obtained from Shanghai Cell Bank, Chinese Academy of Science (Shanghai, China). All cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Hyclone, USA), supplemented with 100 μg/ml streptomycin, 10% heat-inactivated FBS, and 100 U/ml penicillin. All media were incubated in an incubator (Thermo Fisher Scientific Inc., Waltham, MA, USA) (37°C, 100% humidity, and 5% CO₂).

Ovarian cancer SKOV3 and OVCAR-3 cell lines were purchased from the Shanghai Cell Bank of Chinese Academy of Sciences and cultured with RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA USA) supplemented with 10% fetal bone serum (FBS; Hyclone; GE Healthcare; Logan, UT, USA) at 37°C in 5% CO₂. A2780 cells were purchased from Cell Preservation Center of Wuhan University and cultured with Dulbecco's modified Eagle's medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented 10% FBS at 37°C in 5% CO₂. Siva 1 overexpression plasmid pCMV3-Siva 1 or the control pCMV3 [China National Pharmaceutical Group (CNPGC); Sinopharm, Beijing, China] were transfected into A2780 cells with lipofectamine reagent (Invitrogen; Thermo Fisher Scientific, Inc.). A total of 24 h after transfection, 200 µg/ml G418 (Invitrogen; Thermo Fisher Scientific, Inc.) was added to the medium, and the culture medium was renewed every 2 days. Approximately 1 week later, monoclonal cell clusters were observed and selected for culture. Thereafter, the Siva 1 stably expressed A2780 cell line and its control were used for subsequent experiments.