Pm150312

Human gastric cancer lines, including AGS (CL-0022), MKN45 (CL-0292), NCI-N87 (CL-0169), HGC-27 (CL-0107), human gastric epithelial cells GES-1 (CL-0563) and human umbilical vein endothelial cells (HUVECs, CL-0122) were bought from Procell (Wuhan, China). All the cell lines except for HUVECs were cultured in RPMI-1640 media (PM150110, Procell), while HUVECs were maintained in DMEM/F12 basic media (PM150312, Procell) with 10% fetal bovine serum (FBS, 1600044, Gibco, Rockville, MD, USA) and 1% penicillin/streptomycin (PB180120, Procell) at 37°C with 5% carbon dioxide (CO₂).

Alpha mouse liver 12 (AML-12) cell line was established from mouse hepatocytes (source: a 3-month-old CD1 strain MT42 mouse line) that ectopically express human transforming growth factor alpha (ATCC #CRL-2254). AML12 cells were authenticated by their general morphology, short tandem repeat microsatellite marker analysis, capacity to form lipid droplets upon oleic acid treatment and RT-qPCR for mouse genes. AML-12 cells were cultured at 37 °C and 5% CO₂ in DMEM/F12 (Procell #PM150312) supplemented with 10% fetal bovine serum (Procell #164210-50), 10 µg/mL insulin, 5.5 µg/mL transferrin, 5 ng/mL selenium, 40 ng/mL dexamethasone and 1% penicillin/streptomycin (100 U/mL and 0.1 mg/mL, respectively; Procell #PB180120), which is formulated as the AML-12 culture medium (Procell #CM-0602). Cells were seeded to 6-well plates (NEST #703001) or 10-cm dish (NEST #704002). When reaching 80-90% confluence, cells were treated in DMEM (Gibco #11966025) containing various concentrations of glucose (BioReagent grade, Sigma #G7021) for 8 h.

Human breast immortalized cell line MCF-10A and BC cell lines MCF-7, MDA-MB-231 and MDA-MB-468 were provided by ATCC (Manassas, VA, USA) or Procell Bio (Wuhan, China). The culture medium was listed as follow: MCF-10A (DMEM:F12 = 1:1, PM150312, Procell), MCF-7 (MEM, PM150410, Procell), MDA-MB-231 and MDA-MB-468 (Leibovitz's L-15, PM151010, Procell). The medium was supplied with 10% of fetal bovine serum (FBS, FND500, ExCell Bio, Suzhou, China) and 1% Penicillin–Streptomycin Solution (PB180120, Procell). The cells were kept in a cell incubator with 37 °C and 5% CO₂.