蟾毒灵诱导伯基特淋巴瘤细胞凋亡及初步机制研究  

A preliminary study on apoptosis induced by bufalin in human Burkitt lymphoma cells

作　　者：张楠[1] 于文君[1] 韦炜[1] 阎骅[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院血液科,上海200025

出　　处：《内科理论与实践》2015年第3期188-192,共5页Journal of Internal Medicine Concepts & Practice

基　　金：国家自然科学基金项目(项目编号:81170509)

摘　　要：目的 :探讨蟾毒灵(bufalin)对伯基特(Burkitt)淋巴瘤细胞的影响。方法:体外培养人伯基特淋巴瘤细胞株Daudi,以0、5、10、20、40、80 nmol/L浓度蟾毒灵处理细胞,细胞计数试剂盒8(CCK-8)检测细胞抑制率,膜联蛋白(annexin)Ⅴ/碘化丙啶(PI)双染后,流式细胞仪检测细胞凋亡,蛋白质印迹(Western blotting)检测凋亡相关蛋白胱天蛋白酶及B细胞淋巴瘤(Bcl)-2、c-Myc的变化,实时定量(real-time)PCR检测c-Myc RNA水平的变化。结果:CCK-8检测结果显示,蟾毒灵对Daudi细胞增殖有明显的抑制作用,呈药物浓度依赖性,且经48 h处理后的抑制率明显高于24 h,半数抑制浓度(IC50)为(20.03±2.56)nmol/L;流式细胞检测结果表明,蟾毒灵可诱导Daudi细胞发生凋亡;蛋白质印迹检测结果显示,不同浓度蟾毒灵处理Daudi细胞48 h后,胱天蛋白酶3、9及多聚腺苷二磷酸核糖聚合酶1(PARP1)均出现明显剪切,并伴有Bcl-2及c-Myc癌基因的下调;实时定量PCR检测显示,c-Myc癌基因在m RNA水平也出现明显下调。结论:蟾毒灵能够诱导Burkitt淋巴瘤Daudi细胞株发生凋亡,下调的c-Myc及Bcl-2癌基因可能参与了这一过程。Objective To investigate the effects of bufalin on Burkitt lymphoma cells. Methods Burkitt lymphoma cells Daudi were incubated with different concentrations（0, 5, 10, 20, 40, 80 nmol/L） of bufalin in vitro, the inhibition rate was detected by cell counting kit（CCK-8）, apoptosis was detected by annexin Ⅴ/propidium iodide（PI） double staining,apoptotic proteins and B-cell lymphoma（Bcl）-2, c-Myc oncogenes were measured by Western blotting, and m RNA level of c-Myc was determined by real-time PCR. Results CCK-8 test indicated that bufalin significantly inhibited the growth of Daudi cells, the inhibition rate was correlated with the dose, and the half maximal inhibitory concentration（IC50） was（20.03 ±2.56） nmol/L. Results of flow cytometry showed that apoptosis induced by bufalin was in dose dependent man-nerand associated with the activation of caspase families including caspase 3, 9 and polyadenosine diphosphate-ribose polymerase-1（PARP-1）. Furthermore, inhibition of Bcl-2 and c-Myc oncogenes was demonstrated by Western blotting, and real-time PCR showed that m RNA level of c-Myc was inhibited. Conclusions Bufalin can significantly induce apoptosis of Burkitt lymphoma cells, and down-regulation of c-Myc and Bcl-2 may be involved in this process.

关 键 词：BURKITT淋巴瘤 蟾毒灵 细胞凋亡 C-MYC癌基因 

分 类 号：R733.1[医药卫生—肿瘤]