Ccd 841 con

The human colon adenocarcinoma (HT-29) and normal human colon tissue (CCD 841 CoN) cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA). Both cell lines were cultured in the supplier’s recommended basic medium supplemented with 10% fetal bovine serum, 100 units/mL penicillin, and 100 µg/mL streptomycin in an atmosphere of 5% CO₂ at 37 °C. For gene expression analysis, both cell lines were seeded on 100 mm dishes at a seeding density of 2.2 × 10⁶. For the cell viability test, the HT-29 cell line was seeded on a 6-well culture plate at a seeding density of 0.3 × 10⁶.

THP‐1, CCD841 CoN, and HCT116 cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA). THP‐1 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (A4192301, Gibco, USA) supplemented with 10% Fetal Bovine Serum (FBS, Cat # 10100147, Gibco, USA), 1 × HEPES buffer (PYG0019, Boster, China), and 1% Penicillin‐Streptomycin (15140‐122, Gibco, USA). CCD841 CoN and HCT116 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) (Gibco, USA) supplemented with 10% FBS and 1% Penicillin‐Streptomycin. All cells were maintained at 37 °C in a humidified atmosphere containing 5% CO₂. THP‐1 cells were differentiated with 100 ng mL⁻¹ Phorbol 12‐myristate 13‐acetate (PMA, P1585, Sigma‐Aldrich, USA) for 48 h, followed by treatment with 1 µg mL⁻¹ Lipopolysaccharides (LPS, L2654, Sigma‐Aldrich, USA) in the presence or absence of EPA, Orn, and C10 for 48h. CCD841 CoN cells were differentiated with 100 ng mL⁻¹ IL‐1β (Novoprotein, China) and treated with EPA for 48 hours.

Cell lines used in cytotoxicity assays include MCF-7 (human mammary gland adenocarcinoma), HT-29 (human colorectal adenocarcinoma), PC-3 (human prostate adenocarcinoma), CCD 841 CoN (colon epithelial), and HEK-293 (human embryonic kidney) normal cells lines, which were obtained from the American Type Culture Collection (Rockville, MD, USA). All tested cell lines were maintained in a 1:1 mixture of Dulbecco’s modified Eagle’s medium (DMEM) and Ham’s F12 medium, containing 10% heat-inactivated fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 µg/mL) in a humidified atmosphere with 5% CO₂ at 37 °C.