Sgc 7901

The following cancer cell lines were obtained from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China), including human pancreatic cancer cell PANC1, human lung cancer cell A549, human gastric cancer cell SGC-7901, human colon cancer cell HCT116, human liver cancer cell HepG2, human bladder cancer cell T24, and human breast cancer cell MDA-MB-231. Human embryonic kidney cell line HEK293 was a gift from Zhejiang Chinese Medical University and authenticated by STR analysis. All cells were cultured in DMEM supplemented with 10% (v/v) fetal bovine serum (FBS) in an incubator with standard cell culture conditions (37°C, 5% CO₂).

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).

Nineteen cancer cell lines were used for validating experiments in vitro. Four of those were gastric cancer cell lines (NCI-N87, SGC-7901, AGS, and MKN-45), three were melanoma cell lines (MuM-2C, MV3, and A-375), three were hepatocarcinoma cell lines (LM3, 97L, and Huh7), three were thyroid cancer cell lines (KHM-5M, CAL-62, and C643), two were breast cancer cell lines (MCF7 and SK-BR-3), two were pancreatic cancer cell lines (TCC-PAN2 and BxPC3), and two were colorectal cancer lines (DLD-1 and NCIH-747). Cell lines NCI-N87, MuM-2C, LM3, MV3, Huh7, SGC-7901, CAL-62, AGS, MCF7, C643, 97L, SK-BR-3, KHM-5M, A-375, TCC-PAN2, MKN-45, and BxPC3 were purchased from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). Cell lines DLD-1 and NCIH-747 were purchased from The Global Bioresource Center ATCC (Maryland, USA). The cell lines were cultured in RPMI-1640 supplemented with 10% fetal bovine serum in a humidified incubator at 37°C with 95% air and 5% CO2.

GC cell lines MKN-45 and MKN-28, and immortalized human gastric epithelial cell line GES-1, were obtained from 3D Biopharm Biotech Co. Ltd. (Shanghai, China). NCI-N87, HGC-27, AGS, and SGC-7901 cell lines were obtained from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). SNU-216 and GTL-16 cell lines were gifts from the Medical College of Xiamen University (Xiamen, China) and AstraZeneca China R&D Center (Shanghai, China). EBC-1 cell line was obtained from COBIOER BIOSCIENCES Co. Ltd (Nanjing, China). Cell lines were tested and authenticated by short tandem repeat DNA profiling analysis before execution of the experiments. Cells were cultured in Minimum Essential Medium (HGC-27), F12K medium (AGS), Dulbecco's Modified Eagle's Medium (GTL-16), Eagle's Minimum Essential Medium (EBC-1) or Roswell Park Memorial Institute 1640 medium containing 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) and 1% penicillin-streptomycin (Invitrogen, Carlsbad, CA, USA) at 37°C in a humidified atmosphere with 5% carbon dioxide.

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.