摘要
目的:通过体外实验和动物实验,探索华蟾素对白血病细胞株的作用。方法:通过四氮唑蓝(MTT)比色法、Hoechst荧光染色法、原位末端标记(TUNEL)法、DNA凝胶电泳及流式细胞术(FCM)观察华蟾素对人白血病细胞株Jurkat的增殖、细胞凋亡、bcl-2表达的影响;在DBA/2小鼠的腹腔中注射L1210细胞,比较治疗组和对照组小鼠的生存期,评价药物疗效。结果:与对照组相比,1∶100～1∶25浓度的华蟾素能明显抑制Jurkat细胞生长,Hoechst荧光染色可见典型的凋亡形态学改变,TUNEL可见深棕色凋亡细胞,DNA电泳出现凋亡特有的“梯子”式条带,AnnexinV/PI双染色的FCM也证实凋亡的存在。在经1∶100、1∶50、1∶25浓度的华蟾素处理2d后,细胞凋亡率分别为18.78%、24.40%和40.29%。凋亡细胞的bcl-2表达由44.07%下调至16.30%。动物实验中,L1210白血病腹水瘤小鼠经华蟾素治疗后,生存期延长35.90%。结论:华蟾素能抑制Jurkat细胞生长并诱导凋亡,可能是通过下调bcl-2的表达来实现。动物实验证实,华蟾素能延长L1210白血病腹水瘤小鼠的生存期。
Objective To investigate the effect of cinobufacini on leukemia cell line in vitro and in the animal experiment. Methods The effect of cinobufacini on the proliferation, the apoptosis and the bcl-2 gene expression of human leukemia cell line Jurkat was detected using MTT assay, Hoechst immunofluoresence staining, terminal deoxyucleotidyl transferase mediated nick end labeling （TUNEL） assay, DNA agarose gel electrophoresis and flow cytometry. DBA/2 murine injected intraperitoneally with L1210 （lymphatic leukemia cell line）was established as the animal leukemia model. The anti-leukemia effect of cinobufacini was evaluated by comparing the life span between the treatment group and the control group. Results Compared with that in the control group, the proliferation of the Jurkat cell line was inhibited significantly by cinobufacini at the concentration of 1：100-1：25. The morphological changes of the typical apoptosis were observed by Hoecbst staining. The dark brown positive apoptosis cells were found by TUNEL. Agarose gel eletrophoresis of DNA revealed the apoptosis characteristic of “Ladder”. Apoptosis was confirmed by Annexin V/PI double staining using flow cytometry. After the Jurkat cell line was treated with cinobafacini of different concentrations （1：100, 1：50, 1：25） for 2 days, the apoptosis rates were 18.78%, 24.40%, and 40.29%, respectively. The bcl-2 expression was downregulated from 44.07% to 16.3%. In the L1210 leukemia mice model, the life span in the treatment group was 35.9 % longer than that in the control group. Conclusions Cinobafacini can inhibit the growth and induce the apoptosis of the Jurkat cell line might through down-regulating bcl-2 expression. Cinobufacini can prolong the life span of the murine treated with L1210 leukemia cell line.
出处
《诊断学理论与实践》
2005年第4期308-312,共5页
Journal of Diagnostics Concepts & Practice
