Mkn45

SGC7901, BGC823, and MKN45 cells were obtained from the China Infrastructure of Cell Line Resources. NCI-N87, HEK293T, and HEK293FT cells were obtained from the American Type Culture Collection. All cells were maintained and passaged at the State Key Laboratory of Cancer Biology (CBSKL). The MDR SGC7901^ADR and SGC7901^VCR cells were previously established in the CBSKL from SGC7901 cells (23 (link)). All cell lines were authenticated by short tandem repeat analysis and were frequently checked for their morphological features and functionalities. All cell lines were confirmed to be free of mycoplasma contamination. Cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS, glutamine, nonessential amino acids, and antibiotics in a 5% CO₂ incubator at 37°C. To maintain the MDR phenotype, 0.5 μg/mL Adriamycin or 1 μg/mL vincristine was added to the culture medium of SGC7901^ADR or SGC7901^VCR cells, respectively. Unless specified otherwise, cells were treated with rCGA (20 μg/mL) and EGF (50 ng/mL) after serum starvation for 12 hours. Drugs were used as follows: fluorouracil (10 μg/mL), Adriamycin (10 μg/mL), paclitaxel (10 μg/mL), cetuximab (10 μg/mL), erlotinib (20 nM), BFA (5 nM), BMA (1 μM), MG132 (10 μM), PD98059 (5 μM), SB202190 (10 μM), SP600125 (5 μM), and LY294002 (10 μM).

The GC cell lines (MGC803, MKN45, HGC27, SGC7901, and AGS), normal gastric cells GES1, and HEK293T were obtained from the China Infrastructure of Cell Line Resources (Beijing, China). The GC cells were cultured in RPMI-1640 medium while the GES1 and HEK293T were cultured in DMEM medium. All the media were complemented with 10% fetal bovine serum (FBS, Hyclone, Logan, UT, USA), 100 units/mL penicillin, and 100 μg/mL streptomycin (Living, Beijing, China). All the cells were maintained in a humidified incubator containing 5% CO₂ at 37 °C. The cobalt chloride (#232696) and glycine (#V900144) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The formic acid was purchased from Thermo Fisher Scientific (#85178, Waltham, MA, USA). The doxycycline was purchased from Topscience (#T1687, Shanghai, China).

Human gastric carcinoma cell line MKN45 was purchased from China Infrastructure of Cell Line Resources (Beijing, China), and cultured in RPMI supplemented with 20% foetal bovine serum. AGS and N87 cells were purchased from Cell Bank of the Chinese Academy of Sciences (Shanghai, China). AGS cells were cultured in Ham’s F-12K containing 10% foetal bovine serum. The N87 cells were cultured in RPMI 1640 supplemented with 10% foetal bovine serum. The identities of all cell lines used in this study were recently verified by short tandem repeat profiling analysis. The cell lines have recently been tested for mycoplasma contamination.