| Objective: Acute myeloid leukemia(AML)is a malignant clonal disease with abnormal hematopoietic stem cells.The incidence of cancer in the world ranks sixth.With the progress of leukemia therapeutics,the treatment and prognosis of AML have improved greatly.At present,chemotherapy is still the main way to treat acute myeloid leukemia.However,due to the characteristics of bone marrow suppression,cardiac toxicity,drug resistance and easy recurrence of chemotherapy drugs,it is necessary to explore new and more effective drug treatment schemes to improve the efficiency of treatment.Curcumin,as an effective component extracted from plant Curcuma longa,has attracted extensive attention and research due to its inhibitory effect on many cancer cells,including leukemia,but less damage to normal cells.Studies at home and abroad have found that curcumin can inhibit ovarian cancer,lymphoma,leukemia and other tumors,but the study of curcumin on leukemia mostly stays in the observation of the relationship between proliferation and apoptosis,autophagy and proliferation,autophagy and apoptosis are less studied.In this study,KG1 a cells were induced by curcumin alone or in combination with cytarabine,an autophagy inhibitor,3-methyladenine(3-MA),to explore the effects of curcumin and cytarabine alone or in combination on autophagy,proliferation and apoptosis of KG1 a cells.Methods: KG1 a cells were cultured in vitro and pretreated with autophagy specific inhibitor 3-MA.Curcumin and cytarabine alone and in combination acted on KG1 a cells in logarithmic phase for 24 hours.MTT assay was used to detect the proliferation of KG1 a cells in different experimental groups;(2)Annexin V-FITC/PI double-staining flow cytometry was used to detect the apoptotic rate of KG1 a cells in each group;(3)q RT-PCR was used to detect the expression of autophagy-related genes Beclin-1,LC3-II and apoptosis-related genes Caspase-3 and Bcl-2 in KG1 a cells;(4)Western Blot was used to detect the expression of autophagyrelated proteins Beclin-1,LC3-II and apoptosis-related proteins Caspase-3 and Bcl-2 in KG1 a cells;(5)Acridine orange staining and transmission electron microscopy were used to observe the occurrence of autophagy in KG1 a cells.Results:(1)KG1a cells were pretreated with autophagy specific inhibitor 3-MA and then treated with curcumin and cytarabine alone or in combination for 24 hours.The results showed that compared with the untreated group,the activity of KG1 a cells in acute myeloid leukemia decreased significantly(P < 0.05).(2)The apoptotic rate of each drug group pretreated with 3-MA was significantly higher than that of the drug group without 3-MA pretreatment(P < 0.05).(3)The results of q RT-PCR showed that after 3-MA pretreatment,the expression of autophagy-related genes Beclin 1 and LC3-II decreased,the expression of apoptosis-related genes Caspase-3 increased significantly,and the expression of anti-apoptotic gene Bcl-2 decreased significantly in the treatment group,which was significantly different from that in the control group(P < 0.05).(4)Western Blot results showed that the expression of autophagy-related protein Beclin 1 and LC3-II decreased,the expression of apoptosisrelated protein Caspase-3 increased and the expression of anti-apoptotic protein Bcl-2 decreased significantly in 3-MA pretreatment group,compared with the group without 3-MA,the difference was statistically significant(P < 0.05).(5)The results of acridine orange and transmission electron microscopy showed that curcumin promoted the production of autophagosomes in the cytoplasm of KG1 a cells,suggesting that curcumin could induce autophagy of KG1 a cells in acute myeloid leukemia.However,after 3-MA treatment,the number of autophagosomes in the cytoplasm of KG1 a cells treated with curcumin decreased significantly and the autophagy of KG1 a cells was inhibited.Conclusion:(1)Curcumin can significantly reduce the viability of KG1 a cells and induce autophagy.(2)3-MA can inhibit the protective autophagy induced by curcumin and cytarabine,inhibit the proliferation of KG1 a cells,increase the level of apoptosis,and enhance the sensitivity of KG1 a cells to curcumin and cytarabine. |
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