Snu719

Seven human gastric cancer cell lines, MKN45, SNU719, MGC803, AZ521, GT39, SNU16, and MKN28, were obtained from the American Type Culture Collection (ATCC). SNU719, MGC803, AZ521, GT39, and MKN28 cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% (v/v) fetal bovine serum (FBS) and 1% (w/v) penicillin‐streptomycin (Gibco). MKN45 and SNU16 cells were grown in RPMI 1640 supplemented with 10% (v/v) FBS and 1% (w/v) penicillin‐streptomycin (Gibco). All cells were cultured in a humidified incubator at 37°C in a 5% (v/v) CO₂ atmosphere. All human cell lines were mycoplasma‐free and have been authenticated using short tandem repeat (or single nucleotide polymorphism) profiling within the last three years.

Human GC cell lines (SNU719 and SGC7901) were purchased from American Type Culture Collection (Manassas, United States). SNU719 and SGC7901 cells were cultured in RPMI-1640 medium (Gibco) with 10% fetal bovine serum (Gibco). Cells were maintained at 37°C and 5% CO₂. SNU719 and SGC7901 cells were plated onto 6-well plates and reached 70–80% cell confluence on the day of treatment. Cells were divided into the BS group and the control group. The BS group was treated with the FTO inhibitor (Brequinar sodium, V17016, 5 umol/l, InvivoChem), and the DMSO group was treated with an equivalent DMSO concentration as control for 48 h.

Normal human gastric epithelial cells (GES-1) and GC cell lines, including NCI-N87, AGS, SNU-719, MKN-74, MKN-45, SNU-5, and HGC-27, were purchased from the American Type Culture Collection (Manassas, VA, USA). Cells were grown in base Roswell Park Memorial Institute medium (RPMI-1640, Thermo Fisher Scientific, USA) or Dulbecco’s Modified Eagle Medium (DMEM, Thermo Fisher Scientific, USA) supplemented with the addition of 10% fetal bovine serum (FBS, Biological Industries, Israel) and 100 U/mL penicillin/streptomycin solution (Sigma-Aldrich, USA) under standard conditions (5% CO₂ and 37 °C) in humidified incubators.

Gastric epithelial cell lines (GES-1 and HFE145) and gastric cancer cell lines (BGC823, SNU-719, MGC803, AGS, MKN-45 and MKN-28) were acquired from the American Type Culture Collection (ATCC). These cells were cultured in RPMI1640 (Invitrogen) supplemented with penicillin–streptomycin (Invitrogen), GlutaMAX-1 (Invitrogen) and 10% fetal bovine serum (FBS, Gibco).
Sirt7 and ctrl shRNA retroviral particles were purchased from Santa Cruz Biotechnology. The shRNA sequences targeting Sirt7 is shown in Supplementary Table 1. Retrovirus expressing human Sirt7 was generated by sub-cloning Sirt7 cDNA from pcDNA4-Sirt7 plasmid. For retroviral packaging, 293T cells were co-transfected with the retroviral particles. miR-34a was knocked down with the locked nucleic acid (LNA)-antimiR-34a. For transduction, cells were incubated with virus-containing supernatant in the presence of 8 mg/ml polybrene. After 48 hours, infected cells were selected for 72 hours with puromycin (2 mg/ml) or hygromycin (200 mg/ml). For miR-34a treatment, MG803 cells were transfected with miR-34a mimic and miRNA mimic negative control (Ambion, Life Technologies Grand Island) at a final concentration of 200 nM for 96 h using combiMAGnetofection (OZ BIOSCIENCES) in accordance with manufacturer's procedure.

Human GC cell lines AGS, NCI-N87, SNU-719, MKN-45, MKN-74, SNU-1, SNU-5, HGC-27, BGC-823, MGC-803, and SGC-7901, and normal human gastric mucosal cell line GES-1 were obtained from the American Type Culture Collection (ATCC). Cells were grown in basic Roswell Park memorial institute (RPMI-1640) medium (1×) or Dulbecco’s Modified Eagle Medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Biological Industries, Kibbutz Beit-Haemek, Israel), penicillin (100 µg/mL) and streptomycin (100 µg/mL) (Sigma-Aldrich, St. Louis, MO, USA) at 37 °C with 5% CO₂. The antibodies used are listed in Supplementary Table 6.