August 1, 2017
cells were treated with 5 concentrations of each compound (10-fold dilution from 0.001 to 10 mmol/L in duplicates) using the HP D300 digital dispenser (Tecan). Cell viability was measured 48 hours after drug treatment by colorimetric MTS assay following the supplier's recommendations (Promega). Each experiment was repeated at least twice for each cell line, and results were normalized on untreated cells. Curve fitting of doseâresponse data was performed using GraphPad Prism 6 Software, and the two following classical parameters representative of drug sensitivity were derived: (i) the GI50 corresponding to the concentration of drug that inhibits 50% of cell viability and (ii) the AUC corresponding to the area under the doseâresponse curve that provides an overall measure of cumulative response. When the GI50 was not reached, the values were set to the highest concentration tested (10 mmol/L)
Purpose: Tivantinib was initially reported as a selective MET inhibitor and is under phase III evaluation in "MET-high" hepatocellular carcinoma (HCC) patients. However, it has been also proposed as an antimitotic agent. We aimed to evaluate the antitumor effect of tivantinib in HCC cells by combining pharmacologic and molecular profiling.Experimental Design: Sensitivity to tivantinib, JNJ-38877605, PHA-665752, vinblastine, and paclitaxel was tested in a panel of 35 liver cancer cell lines analyzed with exome sequencing, mRNA expression of 188 genes, and protein expression. Drug effect was investigated by Western blot analysis and mitotic index quantification. Expression of candidate biomarkers predicting drug response was analyzed in 310 HCCs.Results: Tivantinib sensitivity profiles in the 35 cell lines were similar to those obtained with antimitotic drugs. It induced blockage of cell mitosis, and high cell proliferation was associated with sensitivity to tivantinib, vinblastine, and paclitaxel. In contrast, tivantinib did not suppress MET signaling, and selective MET inhibitors demonstrated an antiproliferative effect only in MHCC97H, the unique cell line displaying MET gene amplification. HCC tumors with high expression of cell proliferation genes defined a group of patients with poor survival. Interestingly, highly proliferative tumors also demonstrated high MET expression, likely explaining better therapeutic response of MET-high HCC patients to tivantinib.Conclusions: Tivantinib acts as an antimitotic compound, and cell proliferation markers are the best predictors of its antitumor efficacy in cell lines. Ki67 expression should be tested in clinical trials to predict tivantinib response. Clin Cancer Res; 23(15); 4364-75. Š2017 AACR. Š2017 American Association for Cancer Research.
Rebouissou, S; La Bella, T; Rekik, S; Imbeaud, S; Calatayud, AL; Rohr-Udilova, N; Martin, Y; Couchy, G; Bioulac-Sage, P; Grasl-Kraupp, B; de Koning, L; Ganne-CarriĂŠ, N; Nault, JC; Ziol, M; Zucman-Rossi, J;
Journal: Clin. Cancer Res. Pages: 4364-4375