Fig. 5

Combined targeting of c-Met and TrkB kinases is a therapeutic strategy in HCC. (a) HepG2 cells were treated with HGF (10 ng/ml) alone or HGF (10 ng/ml) and BDNF (10 ng/ml) for the indicated time and the activation of c-Met signaling pathway was examined. (b) HepG2 cells were treated with BDNF (10 ng/ml) alone or BDNF (10 ng/ml) and HGF (10 ng/ml) for the indicated time and the activation of TrkB signaling pathway was examined. (c) HepG2 cells were transfected with c-Met siRNA or TrkB siRNA oligonucleotides (d) for 48 h and then treated with BDNF (50 ng/ml) or HGF (20 ng/ml) for the indicated time. The activation of the TrkB and c-Met signaling pathways were examined. (e) HepG2 cells were pre-treated with Crizotinib (0.5 μM) or GNF-5837 (4 μM) (f) for 2 h and then stimulated with BDNF (50 ng/ml) or HGF (20 ng/ml) for the indicated time. The activation of the TrkB and c-Met signaling pathways were examined. (g) HepG2 cells were transfected with the indicated siRNA oligonucleotides and cell growth was measured within 72 h (left panel). The inhibition rate of siRNA oligonucleotides was calculated according to the cell number at 72 h (right panel). (h) HepG2 cells were transfected with the indicated siRNA oligonucleotides for 48 h, and the cell migration activity was examined within 24 h. (i) HepG2 cells were treated with Crizotinib (100 nM) or GNF-5837 (2 μM), as indicated, and the cell growth was measured within 48 h (left panel). The inhibition rate was calculated (right panel). (j) Mice bearing HepG2 cell tumor xenografts were intraperitoneally injected with Crizotinib (16 mg/kg) or GNF-5837 (16 mg/kg) as indicated for 3 weeks. The tumor weights were measured, and the inhibition rate was calculated. Data are shown as the mean ± S.D. and are representative of three independent experiments. **p < 0.01, ***p < 0.001