双氢青蒿素通过激活氧化应激诱导人急性T淋巴细胞白血病细胞凋亡

Dihydroartemisinin Induces Apoptosis of Human Acute T Lymphocytic Leukemia Cells by Activating Oxidative Stress

目的:观察双氢青蒿素(DHA)对人急性T淋巴细胞白血病(T-ALL)Jurkat细胞增殖及凋亡的影响。方法:采用CCK-8法检测DHA对Jurkat细胞的增殖抑制作用及N-乙酰-L-半胱氨酸(NAC)对DHA抑制的细胞活力的的恢复;流式细胞术测定细胞凋亡、活性氧(ROS)产生;Western blot法检测DNA损伤和凋亡相关基因的蛋白表达。结果:DHA对Jurkat细胞增殖有明显抑制作用,具有剂量依赖性(r=-0.936),NAC能部分恢复DHA对细胞增殖抑制的活力。流式细胞术检测显示,随着药物浓度的增高,细胞凋亡率增高(r=0.946),ROS累积量也增高(r=0.965);Western blot结果显示,DHA处理Jurkat细胞后,DNA损伤相关基因γ-H2AX和凋亡相关基因p53、c-Caspase3、BAX、cPARP的蛋白表达明显增加,BCL-2蛋白表达降低。结论:DHA诱导Jurkat细胞产生ROS,引起DNA损伤,激活P53凋亡通路,促使细胞凋亡。

Objective:To investigate the effects of dihydroartemisinin(DHA)on the proliferation and apoptosis of human T-cell acute lymphoblastic leukemia(T-ALL)Jurkat cell.Methods:The effects of DHA on the proliferation of Jurkat cells and the recovery of DHA-inhibited cell viability by N-acetyl-L-cysteine(NAC)were examined by CCK-8 assay.Flow cytometry was performed to analyze the cell apoptosis and generation of reactive oxygen species(ROS).Western-blot was used to detected protein expression of DNA damage-related genes,as well as apoptosis-associated genes,respectively.Results:DHA inhibited the proliferation of Jurkat cells,and shows a concentration-dependent manner(r=0.936),and NAC could partially restore the activity of DHA on cell proliferation inhibition.With the increase of drug concentration,the apoptosis rate(r=0.946)and ROS accumulation was increased(r=0.965).Western blot showed that the protein expressions of DNA damage-related geneγ-H2AX and apoptosis-related genes p53,c-Caspase3,BAX and cPARP were significantly increased,and BCL-2 protein expression was decreased.Conclusion:DHA can induce ROS production in Jurkat cells,which can cause DNA damage,activate the P53 apoptotic pathway,and promote apoptosis of cells.