摘要
本研究探讨藤黄酸(gambogic acid,GA)对K562细胞的抑制作用及机制。以不同浓度的藤黄酸处理K562和K562/A02细胞;用四甲基偶氮唑盐(MTT)法检测GA对细胞增殖的影响;碘化丙啶(PI)染色分析细胞周期;AnnexinV/PI双染法检测细胞的凋亡;JC-1染色法检测线粒体跨膜电位水平;流式细胞仪检测荧光素激活的caspase-3、caspase-8、caspase-9阳性细胞水平。结果显示:藤黄酸呈时间和浓度依赖性抑制K562细胞增殖。K562/A02细胞(GA>2μg/ml)比K562细胞(GA>0.5μg/ml)需要更高的藤黄酸浓度才显示增殖抑制作用。藤黄酸呈浓度依赖性诱导K562细胞凋亡。藤黄酸0、0.5、1.0、2.0μg/ml浓度对K562细胞周期未显示明显影响。藤黄酸作用后的K562细胞的线粒体跨膜电位明显降低。藤黄酸2μg/ml作用24小时激活的caspase 3、caspase 8、caspase 9阳性细胞比例与对照相比分别上升2.19%、-1.95%、34.01%;作用48小时分别上升60.4%、71.3%、77.7%。结论:藤黄酸对K562细胞株有显著的抑制作用,该作用是通过诱导细胞凋亡实现的。藤黄酸不影响K562细胞的细胞周期。藤黄酸通过线粒体跨膜电位途径和胞浆激活途径诱导K562细胞的凋亡。
This study was aimed to investigate the apoptosis-inducing effect of gambogic acid （GA） on K562 cell line and its mechanism. The K562 cells were treated with GA at different concentrations and times, the inhibition rates were detected by MTT assay. Apoptosis induced by GA was assayed by Annexin-V/PI doubling staining. The influence of GA on cell cycle was studied by propidium iodide method. The mitochondrial membrane potential was measured by JC assay. The levels of caspase 3 ,caspase 8 and caspase 9 activated by fluorescein in living K562 cells were measured by caspGLOWTM fluorescein staining kit. The results showed that after incubation with GA, K562 cell proliferation was dramatically inhibited in concentration- and time-dependent manners. K562/A02 cells need higher GA concentration （ 〉 2 μg/ml ） to show antiproliferative effect, compared with that of K562 cells （ 〉 0.5 μg/ml）. Apoptosis could be induced by GA but the influence on cell cycle was not significant. GA could decrease the mitochondrial membrane potential and increase the activated caspase 3, caspase 8, caspase 9 positive cell levels by 2.19%, - 1.95 %, 34.01% in 24 hr and 60.4%, 71.3%, 77.7% in 48 hr respectively. It is concluded that the GA can significantly inhibit the proliferation of K562 cells without influence on cell cycles. The GA triggers K562 cell apoptosis through both intrinsic and extrinsic pathways.
出处
《中国实验血液学杂志》
CAS
CSCD
2009年第6期1443-1447,共5页
Journal of Experimental Hematology
