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 gene γ-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.
题目: 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.
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.