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目的 :研究眼镜蛇毒组分C对白血病细胞的直接作用及机制。方法 :应用MTT、DNA电泳、流式细胞仪、RT_PCR等方法 ,观察眼镜蛇毒组分C对HL6 0等 9株白血病细胞系的毒性作用、量效关系及白血病细胞经过眼镜蛇毒组分C处理后的生物化学、Bcl - 2 /Bax表达水平变化。结果 :眼镜蛇毒组分对 9株人白血病细胞系均有明显的抑制作用 ,IC50 为 0 .0 0 4 6～ 4 .4 0 μg/ml,且呈较好的量效关系 (r为 0 .6 6～ 0 .99)。眼镜蛇毒组分C能诱导HL6 0细胞发生凋亡生物化学变化 ;流式细胞仪分析发现眼镜蛇毒组分C能使一定比例的J6_1、56 2、HL6 0细胞发生凋亡 ,而且凋亡率随蛇毒浓度的增高而增加 ;RT_PCR方法检测发现眼镜蛇毒组分C能使HL6 0细胞Bcl_2基因的表达下调 ,而Bax变化不明显。结论 :眼镜蛇毒组分C对多种白血病细胞株均具有较强的抑制作用 ;此种抑制作用与剂量呈正相关。眼镜蛇毒组分C能诱导白血病细胞发生凋亡 ,此作用与Bcl_2基因的表达下调有关。
Objective: To study the direct effect of component C of cobra venom on leukemia cells and its mechanism. METHODS: MTT, DNA electrophoresis, flow cytometry, RT-PCR and other methods were used to observe the toxic effect of cobra venom component C on 9 leukemic cell lines such as HL60, the dose-effect relationship, and the treatment of leukemia cells with cobra venom component C. Biochemistry, changes in Bcl-2/Bax expression levels. RESULTS: The cobra venom component significantly inhibited 9 human leukemia cell lines with an IC50 of 0.046 to 440 μg/ml and a good dose-effect relationship (r=0. 6 6-0.99). Cobra venom component C can induce biochemical changes of apoptosis in HL60 cells. Flow cytometry analysis revealed that cobra venom component C can cause a certain percentage of J6_1, 562, HL60 cells to undergo apoptosis, and the rate of apoptosis increases. Increased snake venom concentration increased; RT-PCR method found that cobra venom component C can make HL60 cells Bcl_2 gene expression down, while Bax changes are not obvious. Conclusion: Cobra venom component C has a strong inhibitory effect on various leukemia cell lines; this inhibition is positively correlated with the dose. Cobra venom component C can induce apoptosis in leukemic cells, and this effect is related to the down-regulation of Bcl-2 gene expression.