Skov3

The human EOC cell lines (3AO, A2780, and OVCAR3) were purchased from Shanghai Mingjin Biotech Co., Ltd. (Shanghai, China). SKOV3, CAOV3, and HEK293T cells were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The normal ovarian epithelial cell line IOSE-386 was provided by Prof. Jin Zhu (General Hospital of Eastern Theater Command, Nanjing, Jiangsu, China). CAOV3, A2780, HEK293T, and IOSE-386 cells were maintained in Dulbecco's modified Eagle's medium (DMEM; KeyGEN, Jiangsu, China). Specifically, 3AO and OVCAR3 cells were incubated in RPMI-1640 medium (KeyGEN), and SKOV3 cells were cultured in McCoy's 5A medium (KeyGEN). Each culture medium contained 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Waltham, MA, USA), except for the medium used to culture OVCAR3 cells, which contained 20% FBS.

Two human ovarian cancer cell lines (SKOV3, A2780) were purchased from Nanjing KeyGen Biotech.Inc Cells were cultured in α‐MEM medium (Hyclone) supplemented with 10% fetal bovine serum (Israel) and 1% penicillin‐streptomycin (Hyclone), and every 3 days, the medium was changed. All cell cultures were maintained in a humidified incubator (Heraeus) with 5% CO₂ saturation at 37°C, and the cells in their third or fourth passage were used.
To investigate whether IL‐8 played a key role in facilitating cell migration, the exogenous IL‐8 and the inhibitor of IL‐8 receptor (Reparixin) were used. Because the CXCR1/2 binds to IL8 with high affinity and transduces the signal through a G‐protein–activated second messenger system, the CXCR1/2 antagonism (Reparixin, MCE, USA) was commonly used to inhibit the IL‐8 signalling pathway. The cells were divided into four groups: cells without the exogenous IL‐8 or Reparixin but contain the endogenous IL‐8 (Endogenous IL‐8 was secreted by the ovarian cancer cells); cells treated with exogenous IL‐8 (100 ng/mL, MCE, USA) for 48 hours; cells treated with Reparixin (0.1 μmol/L) targeting CXCR1/2 for 48 hours to block the effect of endogenous and exogenous IL‐8; and cells treated with Reparixin (0.1 μmol/L) combined IL‐8 (100 ng/mL) for 48 hours.

EOC cells SKOV3, A2780, and OVCAR3 were purchased from the Cell bank of the Chinese Academy of Sciences (Shanghai, China). SKOV3 cells were cultured in McCoy’s 5A (KeyGen, Nanjing, China) with 10% FBS (Gibco, Grand Island, NY), A2780 cells were cultured in DMEM with high glucose (Gibco, Grand Island, NY) with 10% FBS and OVCAR3 cells were cultured in 1640 medium (Gibco) with 20% FBS.

Human ovarian tumor cell lines SKOV3, OVCAR3, OVCAR5, ES-2 were purchased from Keygen Biotech (Jiang Su, China). HO-8910, HO-8910 PM cell lines were purchased from Cell Bank of the Chinese Academy of Science (Shanghai, China). The HEY cell line was purchased from Shanghai Genechem (Shanghai, China). All cell lines were authenticated by profiling of short tandem repeat analysis. Cells were cultured in RPMI1640 (Invitrogen, Carlsbad, CA) supplemented with 10% fetal calf serum, penicillin (100 U/ml, Gibco) and streptomycin (100 μg/ml, Gibco). Cells were incubated in a humidified atmosphere containing 5% CO₂ at 37 °C and checked regularly for mycoplasma infection.

Five OC cell lines (SW626, SK-OV-3, TOV-112D, A2780, and OVcar3) and a normal ovarian epithelial cell line (IOSE-80) were purchased from Nanjing KeyGen Biotech (Nanjing, China). All the cell lines were initially stored in liquid nitrogen and subsequently allowed to recover in a complete DMEM medium (Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum, 10,000 UI/mL penicillin, and 10 mg/mL streptomycin prior to use. The cells then were cultured in a humidified 37°C incubator containing 5% carbon dioxide.

The human ovarian cancer cell line SKOV3 and human embryonic kidney cell line HEK293T were obtained from the National Collection of Authenticated Cell Cultures (Beijing, China). SKOV3 cells were cultured in McCoy’s 5A medium modified (KeyGEN BioTECH, Jiangsu, China). HEK293T cells were maintained in DMEM medium (Gibco, Carlsbad, CA, USA). All the mediums were supplemented with 10% fetal bovine serum (FBS, Ausbian, Australia) and 100 U/mL penicillin–streptomycin mixture (Solarbio, Beijing, China). All cells were incubated at a temperature of 37 °C under 5% CO₂.