Nugc 3

Seven human GC cell lines (NCI‐N87, MKN74, AGS, NUGC‐3, MKN45, MGC803, and HGC‐27) were obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). All cell lines were maintained in RPMI‐1640 (Gibco BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) except AGS in Ham's F12 medium (Cellgro, Manassas, VA, USA) and incubated at an atmosphere containing 5% CO₂ at 37 °C. Human GC samples and their corresponding nontumorous gastric tissues were collected at the time of surgical resection at The First Affiliated Hospital of Fujian Medical University (Fuzhou, China) from 2008 to 2010. The tissues were immediately frozen in liquid nitrogen and stored at −80 °C freezer or fixed in 10% formalin for paraffin embedding. All samples were collected with patients’ informed consent, and the study was approved by the institutional review board and regulatory authorities of Fujian Medical University. Clinicopathological classification and staging were determined according to American Joint Committee on Cancer seventh edition of GC TNM staging (Wittekind, 2010). No patients had received chemotherapy or radiotherapy before surgery.

The GSE-1, BGC-823, SGC-7901, MGC-803, AGS, MKN-45, NUGC-3 and HEK 293 T cell lines were purchased from Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplementary with 10% FBS, 100 μg/ml streptomycin and 100 U/ml penicillin. All cells were maintained at 37 °C in a humidified atmosphere of 5% CO2.

Six human GC cell lines (NCI-N87, MKN74, AGS, NUGC3, MGC803, HGC-27) were obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). All cell lines were maintained in RPMI-1640 supplemented with 10% FBS except AGS in DMEM/Ham’s F12 medium and incubated at an atmosphere containing 5%CO2 at 37 °C. Gastric adenocarcinoma patients (n = 115) were randomly enrolled in between January 2013 and December 2014 at the Department of General Surgery of the Second Affiliated Hospital of Fujian Medical University. All patients were diagnosed pathologically according to the American Joint Committee on Cancer (AJCC) criteria [26 (link)]. No patient had received chemotherapy or radiotherapy before surgery. Tumor samples and the corresponding noncancerous mucosal tissue were collected from all patients immediately after resection and were frozen in liquid nitrogen. Samples were stored in the Center Laboratory of Second Affiliated Hospital of Fujian Medical University for studies.