Il lβ

MCF-7 and MDA-MB-231 human breast cancer cell lines and DU-145 and PC-3 human prostate cancer lines were from American Type Culture Collection (Manassas, VA, USA) and were maintained according to distributors ' instructions. All cells were grown in a 5% CO₂ atmosphere at 37°C. Pyrrolidine dithiocarbamate (PDTC), SP600125, SB203580, PD98059, and anti-β-actin antibody were purchased from Sigma (St. Louis, MO, USA). Recombinant PTX3, IL-lβ, IL-17, IL-23, IL-34 and TNFα proteins and rabbit anti-human PTX3 antibody for ELISA were from R&D Systems (Minneapolis, MN, USA).

Isolated liver NK cells were cultured with the following cytokines: tumour necrosis factor-α (TNF-α) (50 ng/mL), interferon-γ (IFN-γ) (1 µg/mL), and IL-lβ (25 ng/mL; all from R&D Systems, Minneapolis, MN, USA), in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum (Sanko Chemical Co., Ltd., Tokyo, Japan), 25 mmol/L HEPES Buffer (Gibco, Grand Island, NY, USA), 50 mol/L 2-mercaptoethanol (Katayama Chemical Co., Osaka, Japan), 50 U/mL penicillin, and 50 g/mL streptomycin (Gibco) (hereafter referred to as cRPMI) at 37 °C in a 5% CO₂ incubator. After 24 h incubation, the cultured cells were harvested for further analyses.