Hucct1

BTC cell lines (RBE, HuCCT1, SSP-25, TGBC-24TKB, and TFK-1) were purchased from the RIKEN Cell Bank (Ibaraki, Japan). BTC cell lines (SNU-308 and SNU-1196) were purchased from the Korean cell line bank (Seoul, Korea). RBE, HuCCT1, SSP-25, TFK-1 SNU-308, and SNU-1196 cell lines were grown in RPMI (Roswell Park Memorial Institute) medium supplemented with 10% fetal bovine serum (FBS), penicillin-streptomycin, L-glutamine, and sodium pyruvate. TGBC-24TKB cells were grown in DMEM supplemented with 10% FBS and penicillin–streptomycin. The gemcitabine-resistant cells (SNU-1196-GR and SSP-25-GR cells) were generated in the cell culture system. The cells were treated with a half-maximal inhibitory concentration (IC₅₀) dose of gemcitabine for one month (20 nM for SSP-25 cells and 10 nM for SNU-1196 cells). After one month, the media were replaced with the fresh media containing an IC₉₀ dose of gemcitabine (250 nM for SSP-25 cells and 60 nM for SNU-1196 cells) for another month. After two months, the live cells were identified as gemcitabine-resistant cells. All the cell lines used in this study were tested for mycoplasma contamination and authenticated by STR (short tandem repeat) method. AZD0156 (ATM inhibitor), gemcitabine, and cisplatin were purchased from AdooQ BioScience (Irvine, CA, USA).

Human intrahepatic cholangiocarcinoma cell lines, HuCCT-1 and SSP-25, were obtained from the RIKEN cell bank. All cell lines were cultured in RPMI-1640 medium (FUJIFILM Wako Pure Chemical Corp., Osaka, Japan), supplemented with 10% fetal bovine serum (FBS), in an incubator with 5% CO2 at 37°C.

Six cholangiocarcinoma cell lines used in this study (HuCCT1, SSP-25, RBE, YSCCC, TGBC-24TKB and TFK-1) were purchased from RIKEN Cell Bank (Tsukuba, Japan). The HuCCT1, SSP-25 and RBE cell lines had a KRAS mutation, whereas the remaining three cell lines were KRAS wild type. All in vitro studies were performed at least in triplicate.