双氢青蒿素诱导外周T细胞淋巴瘤Hut-78细胞凋亡及其可能的机制  

Dihydroartemisinin induced apoptosis of peripheral T lymphocytoma Hut-78 cells and its possible mechanism

作　　者：黄琰[1] 吴隼[1] 张媛[1] 字友梅[1] 杨满[1] 郭燕[1] 张灵秀[1] 王莉华[1] 

机构地区：[1]新乡医学院第一附属医院血液科,河南卫辉453100

出　　处：《中国肿瘤生物治疗杂志》2016年第1期57-61,共5页Chinese Journal of Cancer Biotherapy

基　　金：河南省教育厅基金资助项目(No.2006320039)~~

摘　　要：目的：考察双氢青蒿素（dihydroartemisinin,DHA）诱导的外周T细胞淋巴瘤Hut-78细胞凋亡,并进一步研究其作用机制。方法：CCK-8法检测1～30μg/ml DHA对Hut-78细胞增殖的影响,运用Hochest 33258染色技术在共聚集显微镜下观察DHA对Hut-78细胞核形态的影响,流式细胞术检测DHA对Hut-78细胞凋亡的影响,10 mmol/L的活性氧（reactive oxygen species,ROS）清除剂N-乙酰半胱氨酸（NAC）预处理细胞1 h,观察ROS在DHA引起的细胞线粒体膜电位变化中的作用,Western blotting检测在Hut-78细胞凋亡过程中ROS对DHA引起的细胞色素C释放的影响。结果：DHA呈浓度依赖性抑制Hut-78细胞的增殖,并引起细胞核固缩,形成凋亡小体。5、10、20μg/ml DHA引起的细胞凋亡率分别为（25.1±2.8）%、（43.6±3.1）%、（68.9±2.6）%,与对照组相比差异有统计学意义（均P〈0.01）。20μg/ml DHA处理导致Hut-78细胞线粒体膜电位下降（59.4±2.6）%,而经ROS抑制剂NAC预处理之后,线粒体膜电位下降（38.4±2.1）%。DHA能够引起线粒体中细胞色素C的释放,而NAC的预处理能够明显地抑制这一过程,统计结果进一步证明这一点。结论：DHA能够抑制外周T细胞淋巴瘤Hut-78细胞的增殖并诱导细胞凋亡,其作用机制可能与DHA促进ROS依赖的细胞色素C释放有关。Objective ： To investigate dihydroartemisinin （DHA） -induced apoptosis of peripheral T lymphocytoma Hut- 78 cell line and its possible mechanism. Methods： The effect of DHA 1 -30 μg/ml on proliferation of Hut-78 cells was measured by CCK-8 assay. The morphological changes of Hut-78 cell＇s nuclei induced by DHA were observed by Hochest 33258 staining and confocal microscopy. Flow cytometry was used to examine the apoptosis of Hut-78 cells induced by DHA. After pretreating the Hut-78 cells with 10 mmol/L of reactive oxygen species （ROS） scavenger N-acetyl cysteine C （ NAC ） , a role of ROS in changes of mitochondrial membrane potential induced by DHA was evaluated. Effect of ROS on release of cytochondrial C induced by DHA during apoptosis of Hut-78 cells was examined with an immunoblotting assay. Results： DHA inhibited proliferation of the Hut-78 cells in a dose-dependent manner, induced nucleus pyknosis and formed apoptotic bodies in the cells. Apoptosis rates of the Hut-78 cells iuduced with 5, 10 and 20 μg/ml of DHA were （25.1 ± 2.8 ） %, （43,6 ± 3.1 ） % and （68.9 ± 2.6 ） % respectively, which were significantly different from that of the control group （all P 〈 0.01 ）. Furthermore, treatment with 20 μg/ml of DHA droped mitochondrial membrane potential of the Hut-78 cells by （59.4 ± 2.6） % , and pretreating with ROS scavenger NAC droped the mitochondrial membrane potential by （38.4± 2.1 ） %. DHA can induce release of cytochrome C in mitochondrion, which can be significantly inhibited by pretreatment with NAC. Statistic results further demonstrated this point. Conclutions： DHA can effectively inhibit proliferation of the peripheral T lymphocytoma Hut-78 cells and induce their apoptosis. Its action mechanism may be related to ability of HAD to promote release of ROS-dependent cytochrome C.

关 键 词：双氢青蒿素 外周T细胞淋巴瘤 Hut-78细胞 细胞凋亡 活性氧 细胞色素C 

分 类 号：R733.1[医药卫生—肿瘤] R730.54[医药卫生—临床医学]