Mgc 803

AGS, MGC803, SNU16, NCI‐N87, MKN28, MKN45 and KATOIII gastric cancer cell lines, GES‐1 normal gastric epithelial cell line, and HEK293FT human embryonal kidney cell were purchased from National Collection of Authenticated Cell Cultures (NCACC). Cell lines were cultured in indicated mediums supplemented with 10% fetal bovine serum (Gibco): DMEM for HEK293FT cell line, RPMI‐1640 for GES‐1, MGC803, SNU16, NCI‐N87, MKN28, MKN45 and KATOIII cell lines and Ham's F‐12K (Kaighn's) Medium for AGS cell line. Aforementioned cell lines were all authenticated by short tandem repeat (STR) analysis.

Normal human GES-1 cell line and five gastric cancer cell lines (MGC-803, AGS, HGC-27, SGC-7901, and BGC-823) were purchased from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (China). Cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Invitrogen, USA) with 5% fetal bovine serum (Thermo Scientific HyClone, China), in a humidified incubator (Thermo, USA) with 5% CO², 95% air.

Seven gastric cancer cell lines (AGS, MGC80-3, BGC-823, HGC-27, MKN-28, MKN-45, SGC-7901) and a gastric mucosal cell line (GES-1) were obtained from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). HGC27, MKN-28, and MKN-45 cells were cultured in Dulbecco’s Minimum Essential Medium and the other cell lines were cultured in RPMI-1640 Medium, supplemented with 10% fetal bovine serum at 37 °C in a humidified atmosphere containing 5% CO₂. All of the culture media were purchased from Sigma (St. Louis, MO, USA), and fetal bovine serum was purchased from Gibco (catalogue no. 16000-044; Grand Island, NY, USA).

Human gastric cancer cell line MGC-803, human prostate cancer cell line PC3, and human breast cancer cell line Bcap-37 and one normal cell line NIH3T3 were obtained from Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). NIH3T3 was routinely maintained in a DMEM medium, while all the other cell lines were cultured in a 1640 medium. All the cells were grown in the medium supplemented with 10% FBS at 37 °C with 5% CO₂.

A human peritoneal mesothelial cell (HPMC) line, which was established by Prof. Ronco,21 was kindly provided by Prof. You‐Ming Peng (Second Hospital of Zhongnan University, Changsha, China). HPMCs were cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS). Human GC cell lines MKN‐45, MKN‐28, SGC‐7901, MGC‐803, and BGC‐823 were purchased from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). Human GC cells were cultured in RPMI 1640 medium or DMEM supplemented with 10% FBS. All cell cultures were incubated continuously under 5% CO₂ at 37°C. For the GC and HPMC coculture system, transwell chambers (0.4‐μm pores, Corning) separated by a polycarbonate membrane were used similar to the previous experiments performed in our laboratory.6, 22 GC cells (5.0 × 10⁵ cells) were seeded into the top chamber, and HPMCs (1.0 × 10⁵ cells) were placed in the bottom compartment. GC cells had no direct contact with HPMCs, but the soluble factors derived from the GC cell lines could reach HPMCs.