Ovcar 3

Human ovarian cancer cell lines OVCAR-3 and Anglne were provided by the BeNa Culture Collection and Procell Life Science & Technology Co., Ltd, respectively, which were sourced from the American Type Culture Collection. The OVCAR-3 and Anglne cells were routinely cultured in dulbecco’s modified eagle medium (DMEM) containing 10% FBS and 1% penicillin/streptomycin/amphotericin B and were maintained in a humidified incubator at 37°C with 5% CO₂. The cells were subjected to treatment with different concentrations of histamine (1, 5, 10, 50, and 100 ng/mL) and apigenin (0.1, 0.5, 1, 5, and 10 μM) for 48 h. Additionally, 1 μM AZD9496, PHTPP, PPT, and DPN were added to the treatment group and incubation was observed for 48 h.

Three ovarian cell lines, HOSEpiC (human ovarian epithelial cell line, BeNa Culture Collection, Beijing, China), OVCAR-3 (human cancerous ovarian cell line, BeNa Culture Collection, Beijing, China) and HO-8910 (human cancerous ovarian cell line, BeNa Culture Collection, Beijing, China), were purchased and used. HOSEpiC and HO-8910 cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin solution, while OVCAR-3 cells were grown in RPMI 1640 with 20% FBS, 0.01 mg/mL bovine insulin and 1% penicillin/streptomycin solution. These cells were incubated and cultured at 37 °C in a humidified atmosphere of 5% CO₂. For AFM experiments, the ovarian cancer cells were seeded in 35 mm culture dishes with a density of 1 × 10⁴ mL⁻¹ for 24 h. Different concentrations of echinomycin were added into the culture dishes for 3 h of treatment. After the treatments, cells were then washed twice with PBS and immediately used for AFM measurements in 2 mL medium.