小檗胺诱导白血病细胞株NB4细胞凋亡及其机制的研究  

Effects of berbamine on growth of leukemia cell line NB4 and its mechanism

作　　者：何智文[1] 赵小英[2] 徐荣臻[2] 吴东[2] 

机构地区：[1]浙江大学医学院附属第二医院肿瘤研究所,浙江杭州310009 [2]浙江大学医学院附属第二医院血液科,浙江杭州310009

出　　处：《浙江大学学报（医学版）》2006年第2期209-214,共6页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省科技厅科研基金项目(2000KJT012)

摘　　要：目的:探讨小檗胺对白血病细胞系NB4生长的影响及其机制。方法:体外培养人白血病细胞株NB4细胞,经不同浓度小檗胺处理不同时间后,MTT法检测药物对NB4细胞的抑制作用;细胞形态学观察和DNA琼脂糖电泳检测细胞凋亡;流式细胞仪检测DNA含量及细胞周期;巢式PCR及RT-PCR检测PM L/RARα融合基因及Surv iv in基因表达及流式细胞仪检测Caspase3阳性表达率。结果:经不同浓度小檗胺处理不同时间后,NB4细胞生长出现明显的抑制,并呈现明显的时间剂量依赖性,48 h IC50为3.860μg/m l;细胞形态学观察可见细胞凋亡现象,DNA琼脂糖电泳显示典型的凋亡梯形图;流式细胞仪检测发现细胞凋亡率明显增加,经48 h作用后,凋亡率从2.83%增至12μg/m l时的58.44%;虽未发现药物作用后PM L/RARα基因表达量的改变,但检测到NB4细胞经药物作用后Surv iv in基因表达量的减少和Caspase3表达的增加,Caspase3阳性率从对照组的2.06%增至12μg/m l时的70.89%。经相关性分析,随着细胞Surv iv in表达水平的下降和Caspase3表达水平的增高,NB4细胞凋亡率相应地增高。结论:小檗胺对NB4细胞有增殖抑制作用,诱导细胞凋亡可能是其抗肿瘤作用的重要机制之一,但不是通过作用于NB4细胞特异性融合基因PM L/RARα发挥作用的,其抗凋亡的可能分子机制之一是抑制Surv iv in基因的表达和增加Caspase3的表达。Objective： To determine effects of berbamine on the growth of leukemia cell line NB4 and explore its possible mechanisms. Methods： The growth of NB4 cells was examined with MTT assay. Morphological analysis and DNA agarose eleetrophoresis were used to detect apoptosis in NB4 cells, and the apoptosis rate was measured by flow cytometry. The PML/RARα mRNA was determined by nested-PCR, and the Survivin mRNA was tested by RT-PCR. The expression of caspase 3 protein in NB4 cells was evaluated by flow cytometry. Results： The growth of NB4 cells was inhibited significantly after treated with berbamine at different concentrations for different time points, the IC50 value was 3.860 μg/ml at 48 hours. Morphology analysis showed the characteristics of apoptosis, and the DNA agarose electrophoresis showed the typical DNA ladder. The apoptosis rate increased from 2.83% to 58.44% after treated with berbamine at 12 μg/ml for 48 hours. The expression of PML/RARα mRNA presented no significant changes, however, Survivin mRNA was decreased dramatically. The protein expression of Caspase 3 increased significntly from 2.06% to 70.89% after treated with berberine at a concentration of 12 μg/ml for 48 hours. Conclusion： Berbamine could inhibit the growth of leukemia cell line NB4. The induction of cell apoptosis may be one of the mechanisms for suppressing the growth of leukemia cell line NB4. Inhibition of Survivin mRNA and upregulation of Caspase 3 protein might be also involved in cell apoptosis.

关 键 词：小檗胺/治疗应用 白血病/病理学 白血病细胞 NB4细胞 凋亡 机制 

分 类 号：R733.7[医药卫生—肿瘤]