摘要
目的:探讨双氢青蒿素(DHA)对急性髓系白血病(AML)的潜在治疗价值及其作用机制。方法:通过CCK-8检测细胞活力,明确DHA对AML细胞活力的影响;通过荧光探针染色及流式细胞术检测DHA对AML细胞内氧化还原状态的影响;通过免疫印迹技术、腺病毒转染和激光共聚焦分析DHA对细胞自噬水平的影响,并利用不同小分子抑制剂进行实验,进一步探究DHA诱导AML细胞死亡的可能机制。结果:DHA可抑制AML细胞株HL-60和Kasumi-1的活力,同时可显著提高细胞内氧化应激水平,经10μmol/L DHA处理后,HL-60细胞内活性氧含量可升至原来的2.6倍,Kasumi-1细胞内活性氧含量可升至原来的2.0倍。此外,DHA还可上调AML细胞内自噬相关蛋白的表达,增加胞内自噬流,激活自噬。进一步检测发现,自噬抑制剂可降低DHA诱导的细胞死亡,而且通过清除胞内自由基抑制氧化应激水平可抑制自噬的发生进而恢复细胞活力。结论:DHA可通过诱导氧化应激激活AML细胞发生自噬性死亡。
Objective:To investigate the potential value and its mechanism of dihydroartemisinin(DHA)in the treatment of acute myeloid leukemia(AML).Methods:The effect of DHA on the viability of AML cells was detected by CCK-8 assay.The effect of DHA on intracellular oxidation-reduction state was detected by fluorescence probe staining and flow cytometry.Western blot,adenovirus transfection,and laser confocal analysis were used to analyze the effect of DHA on autophagy.The small molecule inhibitors were used to further elucidate the possible mechanism of DHAinduced AML cell death.Results:DHA could inhibit the viability of HL-60 and Kasumi-1 cell lines,and significantly increase the level of intracellular oxidative stress.When treated with 10μmol/L DHA,reactive oxygen species(ROS)in HL-60 cells and Kasumi-1 cells was increased to 2.6 times and 2.0 times,respectively.In addition,the expression of autophagy-related proteins were up-regulated in DHA-treated AML cells,together with the increase of intracellular autophagy flux and activation of autophagy.Furthermore,autophagy inhibitors reduced DHA-induced cell death,and inhibited the level of oxidative stress by scavenging intracellular free radicals,thus inhibiting autophagy and restoring cell viability.Conclusion:DHA can activate autophagic cell death of AML by inducing oxidative stress.
作者
潘巍
沈燕
骆波
曹喻
PAN Wei;SHEN Yan;LUO Bo;CAO Yu(Graduate School,Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang Province,China;Department of Clinical Laboratory,Haiyan People's Hospital,Jiaxing 314000,Zhejiang Province,China;Clinical Laboratory Center,Zhejiang Provincial People's Hospital,Hangzhou 310014,Zhejiang Province,China;School of Laboratory Medicine,Zunyi Medical University,Zunyi 563000,Guizhou Province,China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2022年第4期1011-1017,共7页
Journal of Experimental Hematology
关键词
双氢青蒿素
急性髓系白血病
氧化应激
自噬
dihydroartemisinin
acute myeloid leukemia
oxidative stress
autophagy
