Ges 1

The human normal gastric epithelial cell line GES-1 and the GC cell lines AGS, HGC-27, MGC-803, SGC-7901 and BGC-823 were purchased from Genechem (Shanghai, China). GC cell lines with high metastatic potential (MKN-28 M and SGC-7901 M) and corresponding cell lines with low metastatic potential (MKN-28NM and SGC-7901NM) were constructed from the human GC cell lines MKN28 and SGC-7901, respectively, as described previously [3 (link), 4 (link)]. All cells were cultured in RPMI-1640 medium (GIBCO, Carlsbad, CA, USA) supplemented with 10% FBS, 100 U/ml penicillin sodium and 100 μg/ml streptomycin at 37 °C in a 5% CO2 incubator.

The normal gastric epithelial cell line GES-1 and the GC cell lines AGS, SGC7901, MGC803, and HGC27 were obtained from GeneChem (Shanghai, China). These cells were tested for mycoplasma every month to ensure that they are free of mycoplasma contamination. The method is as follows: the nucleus was stained with DAPI and observed under an oil microscope. Mycoplasma contamination is characterized by radiating or satellite-shaped spots around the nucleus (22 (link)). All cell lines were cultured in RPMI-1640 medium (Corning, Corning, NY, USA), supplemented with 10% fetal bovine serum (FBS) (Clark Bioscience, Richmond, VA, USA), penicillin (100 U/ml), and streptomycin (100 μg/ml) (HyClone, Logan, UT, USA) in a humidified atmosphere at 37°C with 5% CO₂. RPMI-1640 medium lacking Asn was customized by Meilun Biotechnology Co. (Dalian, China). For Asn deprivation assays, the cells were plated overnight in complete RPMI-1640 and subsequently transferred into Asn-free medium after a brief wash with phosphate-buffered saline (PBS).

The normal gastric epithelial cell line GES-1 and gastric cancer cell lines AGS, SGC-7901, MGC-803 and BGC-823 were purchased from GeneChem (Shanghai, China). Cells were cultured in RPMI-1640 (Corning, USA) medium with 10% fetal bovine serum (Gibco, USA), 1% penicillin (100 U/ml) and 1% streptomycin (100 μg/ml) (HyClone, USA). All the cells were cultured in our configured medium, and the transfected cells were cultured in complete medium supplemented with puromycin (HyClone, USA). All the cells were cultured in a humidified incubator at 37°C with 95% air and 5% CO₂.