Effect Of Curcumin Combined With Arsenic Trioxide On Autophagy Of Acute Myeloid Leukemia Cells

Research purpose: Acute myeloid leukemia is a malignant tumor of blood system,which originated from bone marrow and is characterized by malignant proliferation of hematopoietic stem cells.In recent years,the incidence rate has gradually increased and the mortality rate is also high.The purpose of this study is to explore the effects of curcumin combined with arsenic trioxide on the proliferation,apoptosis and autophagy of human acute myeloid leukemia KG1 a cell line,and to analyze the potential mechanism of curcumin combined with arsenic trioxide on acute myeloid leukemia cells,so as to provide new ideas and theoretical basis for clinical treatment of acute myeloid leukemia.Research methods: In this study,human acute myeloid leukemia KG1 a cell line was selected as the research object and cultured in vitro.The cells were divided into control group,curcumin group,arsenic trioxide group,curcumin combined with arsenic trioxide group,3-MA group and curcumin combined with arsenic trioxide group after pretreatment with3-MA for 2 hours.The growth state of cells in each group was observed under the inverted microscope.CCK-8 experiment was used to detect the proliferation inhibition rate of KG1 a cells in each group.The effects of different dosage groups on apoptosis rate were detected by flow cytometry.The fluorescence intensity of cells in each group was observed by dansyl cadaverine staining.The relative expression levels of apoptosis-related proteins Bcl-2,Caspase-3,autophagy-related proteins Beclin-1,LC3-II and PI3K/Akt/mTOR signal pathway-related proteins in each group were determined by Western Blot.Results: The results of CCK-8 experiment showed that curcumin and arsenic trioxide could reduce the proliferation activity of KG1 a cells when they were treated separately for 24 hours,48 hours and 72 hours,and the combined administration of curcumin and arsenic trioxide had a more obvious inhibitory effect on the proliferation of KG1 a cells,and the combined administration of curcumin and arsenic trioxide had a higher inhibitory rate on the proliferation of KG1 A cells.The results of flow cytometry showed that the apoptosis rate of KG1 a cells treated with curcumin combined with arsenic trioxide was significantly higher than that of its single drug group,and both were higher than that of the normal culture control group.Compared with the combined group,the apoptosis rate of the combined group pretreated with 3-MA was higher.The results of dansyl cadaverine staining showed that the fluorescence intensity of KG1 a cells in normal culture group was low,and the fluorescence intensity was weakened under the action of 3-MA.When KG1 a cells were cultured with curcumin,the fluorescence intensity of cells was enhanced,and the cells in arsenic trioxide treatment group also showed similar performance,and the fluorescence intensity of cells in curcumin combined with arsenic trioxide group was significantly higher than that in single drug group.Western Blot results showed that the expression of apoptosis-related protein Caspase-3 in KG1 a cells increased compared with the control group,while the expression of Bcl-2 decreased.The expression of Caspase-3 in the combined group was higher than that in the single drug group and the control group,while the expression of Bcl-2 protein was lower than that in the single drug group and the control group.Compared with the control group,the expression of autophagy-related Beclin-1 and LC3-II proteins in curcumin and arsenic trioxide single-drug group and combined-drug group increased significantly,and the expression of Beclin-1 and LC3-II in the combined-drug group was higher than that in the single-drug group.Compared with the control group,the expression of PI3 K,Akt and mTOR protein in cells of curcumin and arsenic trioxide single drug group and combined drug group decreased,and the expression of this signal pathway protein in combined drug group was lower than that in each single drug group,and the expression level of PI3 K,Akt and mTOR protein in cells of combined drug group pretreated with 3-MA increased significantly.Conclusions:(1)Curcumin and arsenic trioxide can inhibit the proliferation and induce apoptosis of KG1 a cells,and the effect of combined administration is better than that of single drug;(2)Curcumin combined with arsenic trioxide can synergistically up-regulate the expression of autophagy-related proteins Beclin-1 and LC3-II,and improve the autophagy level of KG1 a cells;(3)Curcumin combined with arsenic trioxide may inhibit the proliferation and induce apoptosis of KG1 a cells by inhibiting the molecular mechanism of autophagy induced by PI3K/Akt/mTOR signaling pathway.