Effect of ET-743 on proliferation, cell cycle and apoptosis
. (A) ET-743 inhibits growth in vitro. Cells were treated with escalating doses of ET-743 (0.078-10 nM) for 72 hours and cell growth was evaluated by Cell Titer-Glo® cell viability assay. IC50 values were calculated using the CalcuSyn software, based on the Chou-Talalay method. ICP-2/3: primary cells derived from two intrahepatic cholangiocarcinoma (ICC) patients. TFK-1, WITT and EGI-1: extrahepatic cholangiocarcinoma cell lines; HuH28 ICC cell line; TGBC1: gallbladder carcinoma cell line; KMCH: ICC mixed with hepatocarcinoma cell line. The histograms represent the mean of IC50 values (bars represent SEM) from three independent experiments. Effect of ET-743 on cell cycle (B) and on apoptosis (C)
in vitro. BTC cell lines were treated with 5 nM of ET-743 for 48 h and subjected to cell cycle analysis and apoptosis detection by AnnexinV/PI staining by flow cytometry as described in methods. The bars represent the average with SEM of percentage of cells in each phase (B) and apoptotic cells (C) of three independent experiments. NT: no treated cells.