Snu 16

Twelve gastric cancer cell lines (six were established from male patients: NCI-N87, SNU-484, SNU-1, SNU-638, KATOⅢ, MKN-1; six were established from female patients: SNU-5, MKN-28, SNU-216, SNU-16, AGS, SNU-620) were obtained from Korean Cell Line Bank (KCLB; Seoul, Korea). All of the cells were cultured in RPMI 1640 (Gibco BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) and antibiotics (100 U/mL penicillin and 100 g/mL streptomycin; Gibco BRL).

Nineteen RNA samples from different human normal tissues were purchased commercially (Ambion, Austin, TX). Primary GC tissues were obtained from the Prince of Wales Hospital and Beijing Cancer Hospital and stored at −80 °C. All the samples have obtained Informed consent and the study protocol was approved by Clinical Research Ethics Committee of the Chinese University of Hong Kong and Beijing Cancer Hospital. Eighteen GC cell lines were used in this study. AGS, BGC823, KATOIII, MKN1, MKN28, MKM45, MKN74, N87, and SNU1 cell lines were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA). TMK1, SNU16, SNU638, SNU719, and YCC10 cells were obtained from Korean Cell Line Bank (Seoul, Korea). HGC27, GES1, MGC803, and SGC7901 cells were purchased from Chinese Academy of Sciences (Shanghai, China). All cell lines used in this study have been authenticated and tested for mycoplasma contamination.

Human cancer cell lines (HeLa, MDA-MB-231, A549, HCT15, Jurkat, PC12 and SH-SY5Y) and human fibroblast cell lines (CCL-116 and WI-38) were obtained from American Type Culture Collection (ATCC, Rockville, MD, USA). SNU16, human stomach cancer cell line, was obtained from Korean Cell Line Bank (KCLB, Seoul, Korea). The human epithelial primary cell line F10, and the parental cell line for the drug-resistant mutants, KB-3-1, and multidrug-resistant line, KB-V1 were a generous gift from Dr. Peter Dröge (NTU, Singapore) and Dr. M. Gottesman (NCI, Bethesda, MD), respectively [16] (link). Cell lines were grown in either Dulbecco’s Modified Eagle Medium (DMEM) or RPMI 1640 containing 10% fetal bovine serum, 1% penicillin/streptomycin and maintained at 37°C in a humidified 5% CO₂ chamber. For KB-3-1 and KB-1V, the media was supplemented with an additional 1 mM sodium pyruvate and 10 µg/mL vinblastine, respectively. All resistant lines were incubated in drug-free media prior to cellular proliferation assays.

Cell lines were purchased from Japan Health Science Research Resource Bank (IM95, NUGC3, and MKN1), ATCC (SNU16, AGS) and Korean Cell Line Bank (NCC59, NCC24, SNU1967, SNU719, SNU484, NCC19, and SNU1750). YCC10 and YCC11 (Yonsei Cancer Centre in Seoul, South Korea), GES1 (Dr. Alfred Cheng, Chinese University of Hong Kong), and HFE145 (Dr. Hassan Ashktorab, Howard University) were kind gifts. Cell lines were authenticated using Short Tandem Repeat profiling using ANSI/ATCC ASN-0002-2011 guidelines and tested Mycoplasma negative according to the MycoAlert Mycoplasma Detection Kit (Lonza). Normal human stomach antrum and fundus tissue slides were purchased from Novus Biologicals (NBP2-30203, NBP2-30204). Human tissue microarray slides containing matched normal and gastric cancer cases, and human control tissue sections (liver and muscle) were provided and processed by SingHealth Advanced Molecular Pathology Laboratory, Singapore.

Four human GC cell lines (NUGC-2, SNU-16, SNU-620, and SNU638), obtained from the Korean Cell Line Bank (Seoul, South Korea) were grown in RPMI-1640 medium supplemented with 10% fetal bovine serum and 1% antibiotic solution containing penicillin and streptomycin. Cells were incubated at 37ºC in a humidified atmosphere of 5% CO₂. Genomic DNA was extracted from these cell lines using a QIAamp DNA Mini Kit (QIAGEN).

The human GC cell lines NCI-N87, SNU16, MKN7, MKN28, and AGS were obtained from the Korean Cell Line Bank (Seoul, Republic of Korea) and maintained in RPMI-1640 supplemented with 10% fetal bovine serum. The cells were cultured at 37 °C with 100% humidity and 5% CO₂. LOXO-101, entrectinib, dovitinib, dovitinib lactate, dovitinib dilactic acid, regorafenib, cabozantinib, and crizotinib were purchased from Selleck Chemicals (Houston, TX, USA).