丝氨酸/苏氨酸激酶抑制剂诱导白血病细胞凋亡和自噬分子机制研究

OBJECTIVE This study aimed to identify new molecular targets for leukemia treatment using human leukemia cells as the object to investigate the effect of small molecule serine/threonine kinase inhibitor Danu on the cell activity,cell cycle,apoptosis and autophagy in THP-1 and K562 cells.METHODS The effect of Danu on the viability of THP-1 and K562 cells were examined by MTT assay.The effect of Danu on apoptosis of THP-1 and K562 cells were quantitated by the flow cytometry using an Annexin Ⅴ:7-AAD apoptosis detection kit.The effect of Danu on autophagy in THP-1 and K562 cells,and the intracellular autophagy level were assessed by flow cytometry.The levels of various cellular proteins related to the cell cycle,apoptosis,and autophagy were determined by Western blotting.RESULTS Danu decreased the viability of human leukemia cells,the IC ₅₀ value were 26.89 and 32.51 μmol/L in THP-1 and K562 cells.Danu induced the cell cycle arrest in G ₂ /M phase.The G ₂ /M phase rate of THP-1 cells was 2.9-,3.3- and 3.8-fold compared with the control cells.The G ₂ /M phase rate of K562 cells was 1.2-,3.0- and 9.7-fold compared with the control cells(P<0.01).This study suggested that Danu can induce autophagy via inhibiton of the PI3K/Akt/mTOR signaling pathway in THP-1 and K562 cells.The autophagy level was increased 8.3%,21.5%,20.3% and 22.7%,22.6%,24.9% in THP-1 and K562 cells,compared to control cells,respectively(P<0.01).Danu induced apoptosis via the mitochondria-dependent pathway.The apoptosis level was increased 5.5%,7.7%,10.5% and 13.1%,17.4%,18.2% in THP-1 and K562 cells,compared to control cells,respectively(P<0.01).CONCLUSION This study showed an important theoretical fonction for the application of the drug in the treatment of leukemia.