Effect of Evodiamine Combined with Chloroquine on Hepatocarcinoma

OBJECTIVE   To study the anti-liver cancer effect of evodiamine (EVO) combined with the autophagy inhibitor chloroquine (CQ).METHODS  CCK8 method was used to detect the EVO and EVO+CQ on HepG2 cell vitality. Western blot method was used to detect LC3Ⅱ / LC3Ⅰ, P62, vascular endothelial growth factor (VEGFA) protein expressions. ELISA method was applied to detect VEGFA expression level in the supernatant fluid of HepG2 cells. The effect of cell conditioned medium on HUVEC cell invasion ability was tested.RESULTS  Different concentrations of EVO could significantly inhibit the activity of HepG2 cells in a dose-dependent manner after 24 hours. When combined with CQ, the inhibitory activity was stronger (P < 0.01). Western blot results showed that EVO could up-regulate the expression of LC3Ⅱ/LC3Ⅰ (P < 0.05) and reduce that of P62 (P < 0.05), but show no significant effect on VEGFA, although a downward trend was observed. The results of MDC staining showed that EVO could increase autophagosomes in HepG2 cells. When combining EVO and CQ, the accumulation of autophagosomes in HepG2 cells increased significantly. ELISA results showed that EVO reduced the expression of VEGFA (P < 0.01), and much significantly when combining EVO and CQ (P < 0.01). The HUVECs invasion experiments showed that EVO effectively inhibited the invasion of HUVECs (P < 0.01), and much significantly when combining EVO and CQ (P < 0.01).CONCLUSION  Evodiamine can inhibit the activities of HepG2 cells, increase the autophagy level and reduce the angiogenesis. Evodiamine shows more potent inhibitory activities against HepG2 and angiogensis, when it is combined with chloroquine.