摘要
目的:研究三氧化二砷(ATO)联合槲皮素(Que)诱导人急性髓系白血病(AML)Kasumi-1细胞凋亡的作用,并通过线粒体动力学探讨ATO和Que抗白血病的作用机制。方法:体外培养Kasumi-1细胞,应用CCK-8法筛选最终药物干预浓度和时间,分别将ATO和Que单独、联合干预细胞,流式细胞术检测Kasumi-1细胞凋亡率,透射电镜观察细胞超微结构和线粒体形态学改变,Western Blot法检测Cyt-C、AIF、Mfn-1、Drp-1的蛋白表达水平。结果:ATO和Que均可诱导Kasumi-1细胞凋亡,二者联用细胞凋亡率较对照组显著增加(P<0.05);电镜结果提示药物干预可通过改变线粒体形态学、破坏线粒体超微结构并减少线粒体数量与分布以诱导细胞凋亡;Western Blot结果提示药物联用比对照组及ATO组Cyt-C、AIF、Drp-1蛋白表达水平显著提高,Mfn-1蛋白表达水平显著降低(P<0.01,P<0.05)。讨论:ATO和Que均可通过干预线粒体结构的改变,进而激活诱导线粒体凋亡途径,从而诱导白血病细胞凋亡,二者联用效果更加显著。
To investigate the effect of arsenic trioxide(ATO)combined with quercetin(Que)on apoptosis of human acute myeloid leukemia(AML)Kasumi-1 cells,and to explore the anti-leukemic mechanism of ATO and Que through mitochondrial dynamics.Methods:Kasumi-1 cells were cultured in vitro,and we selected the final concentration and intervening time of drugs by CCK-8 method.ATO and Que was intervenced cells by single or combined action.The apoptotic rate of Kasumi-1 cells was detected by flow cytometry,ultrastructure and mitochondrial morphology were observed by transmission electron microscopy.Protein expression levels of Cyt-C,AIF,Mfn-1 and Drp-1 was detected by Western Blot method.Results:Both ATO and Que could induce apoptosis of Kasumi-1 cells,and the apoptotic rate of both groups increased significantly than the control group(P<0.05).Electron microscopic results indicated that drug intervention could induce apoptosis by altering the morphology of mitochondria,destroying the ultrastructure of mitochondria and reducing the number and distribution of mitochondria.The expression levels of Cyt-C,AIF and Drp-1 in both groups were significantly increased than the control group,and the expression level of Mfn-1 protein was significantly decreased(P<0.01,P<0.05).Conclusion:Both ATO and Que can induce apoptosis by interfering with mitochondrial fusion and mitosis-related proteins,thus inducing leukemia cell apoptosis.The combined effect of ATO and Que is more significant.
作者
沈明月
吴晓龙
崔思远
郭云
王琰
徐瑞荣
SHEN Ming-yue;WU Xiao-long;CUI Si-yuan;GUO Yun;WANG Yan;XU Rui-rong(First Clinical College of Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Institute of Hematology,The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310053,China;Department of Hematology,Affiliated Hospital of Shandong University of TCM,Jinan 250014,China;National Research Center for Assisted Reproductive Technology and Reproductive Genetics,Jinan 250001,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2019年第12期5637-5641,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81173442,No.81473511)
山东省医药卫生科技发展计划项目(No.2013WS0249).
