高三尖杉酯碱诱导K562细胞凋亡的时间节律性及其机制研究  

Time Rhythm of Homoharringtonine Inducing K562 Cell Apoptosis and Its Mechanism

作　　者：卢雪莹[1] 曹维克[1] 叶丽霖[1] 邓之奎[1] 张晓慧[1] 李玉峰[1] 

机构地区：[1]南京医科大学附属淮安第一医院血液科,江苏淮安223300

出　　处：《中国实验血液学杂志》2014年第3期712-716,共5页Journal of Experimental Hematology

基　　金：江苏省淮安市科技支撑计划(HAS2010025)

摘　　要：本研究旨在观察高三尖杉酯碱(HHT)作用于K562细胞后不同时间点凋亡率的变化及其机制。HHT 10ng/ml作用K562细胞后,用MTT法检测不同时间点的细胞增殖活力,Western blot法检测caspase-3凋亡蛋白表达;流式细胞术检测BCL-2蛋白表达;应用透射电子显微镜观测细胞自噬。结果表明,HHT 10 ng/ml处理后的前5 d,K562细胞增殖活性逐渐下降,但在第6天以后增殖活性又逐渐上升;同时激活型caspase-3蛋白从第1天至第7天逐渐增高,而在第8天明显降低(P<0.05);BCL-2蛋白表达水平则呈现先降低后升高(P<0.05)的特点。细胞自噬体在HHT作用第8天明显增多。结论:HHT作用后,K562细胞凋亡水平变化呈现先升高后下降的特点,其机制与HHT作用后期K562细胞自噬水平升高有关。This study was aimed to explore the change of K562 cell apoptosis at different time point after homoharringtonine （HHT） treatment and its mechanism. After treatment of K562 cells with 10 ng/ml HHT, the cell viability was tested with M＇IT assay; the expression of caspase-3 was detected with Western blot; the BCL-2 expression was analyzed with flow cytometry; the autophagosome was observed by electron microscopy. The results showed that the viability of K562 cells reduced gradually from day 1 to day 5 and ascended from day 6 to day 8 after HHT treatment. At the same time, the cleaved caspase-3 expression level of K562 cells increased gradually from day 1 to day 7, but reduced at the day 8 （ P 〈 0.05 ）. From day 1 to day 8 after HHT treatment, the BCL-2 expression level declined firstly and then went up （ P 〈 0.05 ）. Autophagosome was also seen remarkably at day 8 after HHT treatment. It is concluded that the apoptosis level of K562 cells after being treated with HHT enhances firstly and then declines , which may be associated with higher autophage level in the late stage of HHT treatment.

关 键 词：高三尖杉酯碱 K562细胞 CASPASE-3 BCL-2 

分 类 号：R733.7[医药卫生—肿瘤]