Evodiamine activates autophagy and inhibits the proliferation in colon cancer cell / 中草药

ABSTRACT

Objective To investigate the effect of evodiamine (Evo) on the autophagy and proliferation of colon cancer HCT-116 cells and the underlying mechanism. Methods The effect of Evo on proliferation of HCT-116 cells was detected by CCK-8 method. After being processed with Evo (3 and 6 μmol/L) for 48 h, the number of autophagic vesicles were detected by MDC method. The amount of ROS in HCT-116 cells was measured by DHE assay, and the protein related with autophagy and AMPK/mTOR pathway was detected by Western blotting. After the treatment of Evo (6 μmol/L) combined with autophagy inhibitor 3-MA (3-methyladenine) or apoptosis inhibitor Z-DEVD-FMK respectively for 48 h, Western blotting was used to detect the expression of autophagy and apoptosis-related protein in HCT-116 cells. Results Compared with the control group, Evo inhibited the proliferation of HCT-116 cells in a dose-dependent manner; After treated with Evo (3 and 6 μmol/L) for 48 h, the amount of intracellular ROS and autophagic vesicles were increased, the protein expression levels of LC3, p-AMPK, and mTOR were increased while the expression of p62 was increased. After being treated with Evo and autophagy inhibitor, the protein expression of LC3 was decreased while activated Caspase-3 was increased; Combination of Evo and apoptosis inhibitor increased the expression of LC3 and inhibited the expression of activated Caspase-3. Conclusion Evo can activate autophagy of HCT-116 cells through AMPK/mTOR pathway and inhibit the proliferation, and the effect of autophagy and apoptosis on cells are complementary.