Mgc 803

Gastric cancer cell lines, MGC-803 (poorly-differentiated adenocarcinoma) and MKN28 (well-differentiated adenocarcinoma) were purchased from the ATCC (Manassas, VA, USA). The cells were maintained in RPMI-1640 media, supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin in a humidified atmosphere of 5% CO₂ at 37°C. All cells were harvested by centrifugation, rinsed with phosphate buffered saline (PBS), and subjected to total protein and RNA extraction. We exposed cells to SAHA and MG132 for the following experiments.

Cell viability was determined with the MTT method [19 (link), 20 (link)]. The human hepatocellular carcinomas cells (HepG2), the human colorectal carcinoma cells (HCT-116) and the human gastric carcinoma cells (MGC-803) were purchased from ATCC. HepG2, MGC-803 and HCT-116 were respectively cultured in DMEM and RPMI-1640 mediums, which were supplemented with 10% fetal bovine serum at 37 °C in a humidified atmosphere containing 5% CO₂. HepG2, HCT-116, and MGC-803 cells (1 × 10⁴) were seeded in 96-well tissue culture plates. Cells were treated in triplicate with five concentrations (50, 25, 12.5, 6.25 and 3.125 μM) of the tested compounds for 24 h, with 5-fluorouracil (5-FU) as positive control. Subsequently, 100 μL of MTT (5 mg/mL) was added and the cells were incubated for additional 2.5 h. Thereafter, the supernatant was discarded and 0.15 ml of DMSO was added to each well, then the plate was mixed on a microshaker for 10 min and read on a microplate reader at 490 nm.

The non-malignant human gastric mucosal epithelial cell line GES-1 (RRID: CVCL_EQ22) and human GC cell lines (MGC-803, RRID: CVCL_5334; SGC-7901, RRID: CVCL_0520) were purchased from ATCC. Cells were cultured in RPMI-1640 medium (Gibco, United States) with 10% fetal bovine serum (Wisent, China). The cells were cultured at 37°C in an incubator supplemented with 5% CO₂.

Three GC cell lines (AGS, MKN45 and MGC803) were obtained from ATCC. Normal gastric epithelial cell line GES-1 was preserved in our laboratory. GC cell lines were cultured in 1640 medium (11,879,020, Thermo, USA), supplemented with 10% fetal bovine serum (10101145C, Thermo, Australian, USA), 100 units/mL penicillin and 0.08 mg/mL streptomycin. All cells were incubated in a 5% CO₂ atmosphere at 37°C.

Cell lines HEK293T, GES-1, LO2, PANC-1, MGC-803, HepG2, HT-29, U251, MCF-7, H1299, ACHN, NCM460, CRYPT, LoVo, HCT-116 and RKO were all obtained from ATCC. Cell lines were routinely cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Hyclone) or high glucose Dulbecco’s Modified Eagle Medium (DMEM) medium (Hyclone), which contained 10% fetal calf serum (Gibco), 100 U/mL penicillin, and 100 U/mL streptomycin, in a humidified cell incubator at 37 °C with an atmosphere of 5% CO₂.