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23 protocols using snu 1

1

Gastric Cancer Cell Line Maintenance

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Human gastric carcinoma cell lines SNU-1 (KCLB no.00001.1), SNU-216, (KCLB no.00216), SNU-601 (KCLB no.00601), MKN-45 (KCLB no. 80103), and AGS (KCLB no.21739) were obtained from the Korean Cell Line Bank (KCLB) of Seoul National University (Seoul, South Korea). Mycoplasma testing was performed for all cell lines used, and mycoplasma were eliminated using MC-210 (WakenBtech Co., Ltd). Cells were maintained in DMEM (Thermo Fisher Scientific, Inc.) containing 10% FBS, sodium bicarbonate (Sigma-Aldrich; Merck KGaA), sodium pyruvate (Gibco; Thermo Fisher Scientific, Inc.), and antibiotics (50 U/ml penicillin and 50 µg/ml streptomycin; Gibco; Thermo Fisher Scientific, Inc.) in a 5% CO2 humidified incubator at 37°C. The culture medium was refreshed every 2–3 days. Mitogen-activated protein kinase (MAPK) inhibitor PD98059 and phosphoinositide 3-kinase (PI3K) inhibitor LY294002 were obtained from Sigma-Aldrich; Merck KGaA. To block the EphA2 function of gastric cancer cells, a novel EphA2 receptor inhibitor ALW-II-41-27 (MedChem Express) was used. For all in vitro studies, ALW-II-41-27 was dissolved in 0.01% DMSO and then diluted to a final concentration of 1 µM.
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2

Characterization of Human Gastric Cancer Cell Lines

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Human gastric cancer cell lines (SNU-1, -5, -16, 216, -484, -601, -620, -638, -668, -719, AGS, MKN45, KATO-III, and N87) were purchased from the Korean Cell Line Bank (Seoul, Korea). The identities of the cell lines were authenticated by DNA fingerprinting analysis [19 (link)]. All cell lines were banked and passaged for less than 6 months before use. The cells were cultured in RPMI 1640 medium (Thermo Fisher Scientific Inc., Waltham, MA) supplemented with 10% fetal bovine serum (Welgene, Daegu, Korea) and 10 μg/mL gentamicin (Cellgro, Manassas, VA) at 37°C and 5% CO2.
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Gastric Cancer Cell Line Cultivation

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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).
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Gastric Adenocarcinoma Cell Line Cultivation

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Human gastric adenocarcinoma cell lines AGS, KATO-III, MKN-45, and SNU-1 were purchased from Korean Cell Line Bank (Seoul, Korea). All four cell lines were maintained in RPMI-1640 (Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Invitrogen), 100 U/mL penicillin G and 100 μg/mL streptomycin, and were maintained at 37°C in a humidified atmosphere of 95% air and 5% carbon dioxide. Astaxanthin was purchased from Sigma-Aldrich (St. Louis, MO, USA) and was dissolved in dimethyl sulfoxide (DMSO) solution and stored at 4°C.
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5

Culturing Human Gastric Cancer Cell Lines

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We purchased three human gastric cancer cell lines SNU-1, SNU-216, and SNU-484 from the Korean Cell Line Bank (Seoul, Korea). All cells were tested for mycoplasma contamination and were maintained in Roswell Park Memorial Institute (RPMI) medium supplemented with 10 % fetal bovine serum (FBS). The cells were cultured in a 5 % CO2 incubator at 37 °C.
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6

Profiling Methylome and miRNome in GC

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To profile the methylome and miRNome of GC, 3 GC and adjacent normal tissue samples were obtained, along with informed consent, from Pusan National University Yangsan Hospital in Korea. EpCAM+/CD44+ GC cells were isolated from the 3 GC tissues according to a previously described protocol.10 To measure the protein expression levels of miR‐1271 target genes in GC tissues, 3 paired normal tissues and GC tissues were obtained, along with informed consent, from Chungnam National University Hospital in Korea. These studies were approved by the Internal Review Board at the corresponding hospitals, and all the experiments were performed in accordance with the relevant guidelines and regulations.
Nine GC cell lines (SNU‐1, SNU‐216, SNU‐484, SNU‐601, SNU‐620, AGS, KATO III, MKN1, and MKN74) and 293T cells were purchased from Korean Cell Line Bank (Seoul, Korea). The GC cell lines and 293T cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium and Dulbecco’s Modified Eagle’s Medium (DMEM, WELGENE, Daegu, Korea), respectively, supplemented with 10% fetal bovine serum (FBS) (HyClone, Logan, UT, USA) in a CO2 incubator, and the cell lines were authenticated by short tandem repeat (STR) DNA profile analysis performed by the Korean Cell Line Bank facility.
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7

Gastric Cancer Cell Lines Culture

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The human gastric cancer cell lines AGS (wild-type p53) and SNU1 (wild-type p53) were purchased from the Korean Cell Line Bank (Seoul, Korea) and cultured in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% fetal bovine serum (FBS; Gibco, MD, USA) and 1% antibiotic-antimycotic solution. Cells were incubated at 37°C with 5% CO2. Econazole, fluconazole, miconazole, and Z-VAD-FMK were purchased from Selleckchem (TX, USA). The reagents were diluted in dimethyl sulfoxide (DMSO) to obtain a final concentration of 100 mM.
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8

Culturing Human Cancer Cell Lines

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Human gastric cancer cell line (SNU-1, obtained from the Korean Cell Line Bank, Seoul, Korea) and human colon cancer cell line (HT29, obtained from ATCC, Rockefeller, Massachusetts, USA) were maintained in Roswell Park Memorial Institute (RPMI) 1640 medium. All mediums were supplemented with 10% fetal bovine serum (GIBCO, Grand Island, New York, USA), penicillin (100 unit/mL) and streptomycin (100 µg/mL). Cells were incubated in a humidified atmosphere at 37°C with 95% air and 5% CO2.
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9

Maintenance of Gastric and Endothelial Cells

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The human GC cell lines, MKN45, MKN74, SNU1, SNU216, SNU668, AGS, and NCI-N87 were purchased from the Korean Cell Line Bank (KCLB) and were maintained at 37 °C in a humidified atmosphere containing 5% CO2, in RPMI 1640 (Hyclone, South Lagan, UT, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin. Human umbilical vein endothelial cells (HUVECs) were purchased from the American Type Culture Collection (ATCC) and were cultured in endothelial cell medium (ECM; ScienCell, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere containing 5% CO2. All cell lines were routinely tested for mycoplasma contamination.
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10

Human Gastric Carcinoma Cell Lines

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Human gastric carcinoma cells AGS, KATO-III, MKN-1, MKN-28, MKN-45, MKN-74, N87, SNU-1, SNU-5, SNU-16, SNU-216, SNU-484, SNU-601, SNU-620, SNU-638, SNU-668, and SNU-719 were purchased from the Korean Cell Line Bank (Seoul, Korea). YCC-1, YCC2, YCC-3, and YCC-7 were kindly provided by Dr. Hyun Cheol Chung (Yonsei Cancer Center, Seoul, Korea). OCUM-2M was kindly provided by Dr. Masakazu Yashiro (Osaka City University, Osaka, Japan). YCC-1, YCC-2, YCC-3, and YCC-7 were maintained in Dulbecco’s modified Eagle’s medium (Gibco-BRL, Carlsbad, CA) supplemented with 10% heat-inactivated FBS, 100 U/ml penicillin, 100 U/ml streptomycin, and 2 mM glutamine. All other cell lines were cultured in RPMI-1640 medium (Gibco-BRL) supplemented with 10% heat-inactivated FBS, 100 U/ml penicillin, 100 U/ml streptomycin, and 2 mM glutamine. All cells were incubated in a humidified atmosphere contained 5% CO2 at 37°C.
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