Skov3

Human melanoma cell line A375, ovarian cancer cell line SK-OV-3, lung large cell carcinoma cell NCI-H460, squamous cell carcinoma of head and neck cell CAL-27 were purchased from the National Infrastructure of Cell Line Resource (Beijing, China). Breast cancer cell line MDA-MB-231, human T lymphocyte leukemia cell Jurkat E6, human embryonic kidneys cell HEK-293T, and murine colon carcinoma cell MC38 were the storage of our laboratory. Murine breast cancer cell line 4T1 was from the American Type Culture Collection (ATCC, MD, USA).
The A375, MDA-MB-231 and CAL-27 were cultured in the Dulbecco's Modified Eagle's Medium (Invitrogen, CA, USA) with 10% fetal calf serum (FBS, Invitrogen), penicillin (100 U/mL) and streptomycin (100 μg/mL) (Invitrogen). The SK-OV-3 was cultured in the McCoy's 5A Media (Invitrogen) with 10% FBS, penicillin and streptomycin (P/S, Invitrogen). The NCI-H460, Jurkat E6, MC38 and 4T1 were cultured in the RPMI-1640 medium (Invitrogen) supplemented with 10% FBS and P/S. All cells were cultured at 37 °C and 5% CO₂.
The A375, SK-OV-3, MDA-MB-231, NCI-H460 and CAL-27 cells were seeded into 12-well plate with full growth medium at 2 × 10⁵ cell per well for 24 h before treatment and cultured overnight. The cells were treated with IFN-γ (10 ng / mL) and different concentrations of CGA (0 - 200 μM) for 48 h.

Human colorectal cancer cells HT29 and human breast cancer cells DU4475 were purchased from ECACC (Porton Down, UK). The human eosinophilic leukemia cell line EOL-1 was purchased from DSMZ (Braunschweig, Germany) and the human ovarian cancer cell line SKOV3 was from ATCC (Manassas, USA). HOS osteosarcoma cells, Molm13 acute myeloid leukemia cells and MV3 melanoma cells (all human) were kindly provided by the Departments of Pediatric Hematology and Oncology, Oncology and Dermatology, respectively (all University Medical Center Hamburg-Eppendorf, UKE). The human pancreatic cancer cell line PaCa5061 was provided by the Department of General, Visceral and Thoracic Surgery at UKE (Kalinina et al. 2010 (link)). The human gastric cancer cell line GC5023 was newly established in the framework of this study (see the next paragraph).
All aforementioned cell lines, except SKOV3, were grown in RPMI-1640 medium with 2 mM L-glutamine, supplemented with 10% fetal calf serum (FCS) and 1% penicillin (50 U/mL) and streptomycin (50 μg/mL) (all from Thermo Fisher Scientific, Waltham, USA), at 37°C with 95% H₂O-saturated atmosphere and 5% CO₂. SKOV3 cells were cultured in McCoy’s 5A medium containing 10% FCS, 2 mM L-glutamine and 1% penicillin/streptomycin (P/S) (all from Thermo Fisher Scientific).

OC cell lines (SKOV3 and A2780) were purchased from the Chinese Academy of Sciences Cell Bank (Shanghai, China). SKOV3 (the IC₅₀ for DDP is 5.1 μM) and SKOV3/DDP (the IC₅₀ for DDP is 52.7 μM) cells were cultured in Dulbecco's Modified Eagle Medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% foetal bovine serum (FBS; Invitrogen, Carlsbad, CA, USA) supplemented with 1% penicillin and streptomycin. A2780 (the IC₅₀ for DDP is 4.8 μM) and A2780/DDP (the IC₅₀ for DDP is 42.4 μM) cells were cultured in RPMI-1640 (Gibco) containing 10% FBS and 1% penicillin and streptomycin. DDP (6 μM) was added to the medium used for culturing SKOV3/DDP and A2780/DDP cells to maintain resistance. All cells were cultured in a humidified incubator at 37 °C in an atmosphere of 5% CO₂.