尖吻蝮蛇蛇毒抑瘤组分1诱导白血病K562细胞凋亡的线粒体机制

本研究探讨尖吻蝮蛇毒抑瘤组分1(component1from Agkistrodon acutus venom,AVVC-1)诱导人慢性髓系白血病(CML)K562细胞线粒体凋亡途径。选择K562细胞株为实验对象,采用Jc-1检测细胞线粒体的膜电位,Western blot检测线粒体内细胞色素C蛋白的表达,免疫荧光检测细胞色素C蛋白的分布。结果表明,不同浓度梯度AVVC-1(12.5,25,50,100μg/ml)处理6 h后,K562细胞线粒体膜电位明显下降(P<0.01)。AVVC-1 30μg/ml处理48 h后,K562细胞线粒体内细胞色素C蛋白表达量明显下降(P<0.05),伴随胞浆内细胞色素C的荧光强度增强。结论:尖吻蝮蛇毒抑瘤组分1可以促使细胞色素C的释放并激活线粒体死亡途径,从而诱导K562细胞发生凋亡,并由此发挥其抗肿瘤活性。

Mitochondrial Mechanisms of Apoptosis of Human Leukemia K562 Cells Induced by AVVC-1

This study was purposed to investigate apoptosis pathway of leukemia K562 cells induced by anticoagulant fraction from Agkistrodon acutus venom （AVVC-1）. The mitochondrial transmembrance potential （Am） of leukemia K562 cells was detected by flow cytometry with JC-1 single staining. The expression of cytochrome C in the mitochondrial of leukemia K562 cells was analyzed by Western blot after AVVC-1 treatment. The distribution of cyto- chrome C in leukemia K562 cells was measured by immuno-fluorescence test. The results showed that the potential of mitochondrial membrane decreased after treatment with different concentrations of AVVC-1 （ 12.5, 25, 50, 100 μg/ml） for 6 h （ P 〈 0.01 ）. The expression level of cytochrome C protein in mitochondria obviously declined after treatment with 30 μg/ml AVVC-1 for 48 h, and the fluorescent intensity of cytochrome C in cytosol was enhanced at the same time. It is concluded that AVVC-l-induced K562 cell apoptosis is related with mitochondrial damage, and cytochrome C may be a useful agent for investiatin human leukemia therapy by usin AVVC-1.