Fig. 6

Anti-proliferative effects of the combination treatments in vivo. HCC xenograft model with treated with control (vehicle-treated mice), biochanin A (25 mg/kg/day), SB590885 (7.5 mg/kg/day) and the combination for 5 weeks. The size (a), volume (b) and weight of the tumours (c) were significantly reduced by the combination treatment compared to those of the control or single treatments. No significant differences in body weight were observed at the end of the treatment period (d). Immunohistochemical analyses of the xenograft tumours revealed that the biochanin A and SB590885 combination effectively inhibited the expression of PCNA, a marker of tumour proliferation (e). Western blot analysis of the xenograft tumours revealed that the combination of biochanin A and SB590885 inhibited activation of the ERK MAPK and PI3K/AKT/mTOR signalling pathways in the in vivo xenograft model (f). The results are the mean ± SD of three independent experiments performed in triplicate. **P < 0.001 versus single treatments (one-way ANOVA followed by a Student–Newman–Keuls test)