【作者】 朱志图； 侯科佐； 刘云鹏； 何莉； 张敬东； 李丹；

【Author】 Zhu Zhi-Tu Hou Ke-Zuo Liu Yun-Peng Department of Medical Oncology, The first Affiliated Hospital, China Medical University, Shenyang 110001, China

【机构】 中国医科大学附属第一医院肿瘤内科；

【摘要】 目的:研究蟾蜍灵与全反式维甲酸(ATRA)联合对急性早幼粒细胞白血病(APL)细胞株NB4细胞的诱导分化作用。方法:采用锥虫蓝拒染法,显微镜下观察形态变化, 计算不同浓度下细胞生存率(细胞生存率=加药组存活细胞数／对照组存活细胞数×100％),绘制增殖曲线。Wright- Giemsa染色形态学分析及流式细胞仪进行DNA含量检测细胞凋亡。细胞诱导分化的检测:(1) 形态学分析:采用 Wright-Giemsa染色,在显微镜下观查形态学变化。(2) NBT 还原试验:计数200个细胞,计算NBT阳性百分比。(3)细胞表面标记抗原CD11b检测:采用流式细胞仪检测。结果: 蟾蜍灵以时间、剂量依赖的方式抑制NB4细胞增殖,24h、48h 和72h抑制细胞增殖50％的药物浓度(IC50)分别为51nmol／ L,35 nmol／L,17 nmol／L。蟾蜍灵20nmol／L处理24h, 形态学观察NB4细胞出现典型的凋亡形态学变化,流式细胞仪解析12h、24h及48h凋亡形成的亚二倍体(sub2N)细胞分别为13．7％,20．5％及37．3％。蟾蜍灵5 nmol／L与ATRA30 nmol／L联合明显诱导NB4细胞向成熟粒细胞分化、联合应用较单独应用ATRA时NBT还原率增加了59．7％及CD11b 表达增加了22．4％结论:蟾蜍灵以时间、剂量依赖的方式抑制NB4细胞增殖,高浓度时诱导NB4细胞凋亡。蟾蜍灵与ATRA联合应用可显著增强ATRA对NB4细胞的诱导分化作用。更多还原

【Abstract】 Objective: To investigate the effect of bufalin combined with all-trans retinoic acid-induced (ATRA) differentiation of acute promyelocytic leukemia (APL) cells NB4 in culture. Mathods :NB4 cells were serial subcultivation .Cell viability was determined by trypan blue dye exclusion. The apoptosis of NB4 cells was assessed by morphological analysis. Differentiation of NB4 cells was assessed by morphological analysis and the nitro blue tetrazolium(NBT) reduction test and expression of the granulocyte/macrophage-specific antigen CD11b. Raaults: Bufalin combined with ATRA can induce differentiation of NB4 cells towards mature stages, NBT reduction was increased 59.7% and CD11b expression was also increased 22.4% in combination of bufalin and ATRA were higher than that of ATRA alone , while the concentration of ATRA needed in the combination was reduced to 30% and the time of differentiation was reduced from 7 days to 4days. Conclusion: The combination of ATRA with bufalin can significantly enhance the differentiation of NB4 cells in vitro culture by ATRA.更多还原