A549 세포에서 5-nitro-5’-hydroxy-indirubinoxime의 apoptosis과 autophagy 유도 효과

Abstract

BACKGROUND: Our group recently demonstrated that 5-nitro-5'-hydroxy-indirubin-3'-oxime (5-OH-NIO), a novel Indirubin-3'-oxime derivative as potent CDK inhibitors. In this study, the effect of 5-OH-NIO was examined in A549 human non-small lung cancer cells. METHODS: Cell viability was examined by MTT assay. Cell cycle were examined by Flow cytometry and apoptosis was measured using annexin V/PI staining and immunoblot. Autophagy was characterized by the increase in LC3B-II and the formation of autophagosome and acidic vesicular organelles (AVOs). RESULTS: 5-OH-NIO inhibited the proliferation of A549 cells in a dose-dependent manner. Cell cycle analysis showed the G2/M arrest in 5-OH-NIO-treated cells. 5-OH-NIO increased the number of apoptotic cells by intrinsic apoptotic pathway. 5-OH-NIO also induced autophagy in A549 cells evidenced by the increased levels of LC3 type II expression and the accumulation of AVOs. The inhibition of autophagy enhanced 5-OH-NIO-mediated apoptosis. CONCLUSIONS: These results suggest that 5-OH-NIO exerts anti-tumor effects by inducing apoptosis and autophagy, and autophagy inhibition enhances 5-OH-NIO-mediated apoptosis in A549 cells.