尖吻蝮蛇毒抑瘤组分Ⅰ通过促进半胱天冬酶3表达诱导K562/A02细胞凋亡

Component Ⅰ from Agkistrodon Acutus Venom Induces Apoptosis of K562/A02 Cells by Promoting Caspase 3 Expression

本研究探讨尖吻蝮蛇毒抑瘤组分Ⅰ(component I from Agkistrodon acutus venom,AAVC-Ⅰ),对人慢性髓系白血病(CML)细胞生物学特性的影响。选择K562/A02细胞株为对象,培养体系中加入不同浓度的AAVC-Ⅰ(6.25-100μg/ml),用CCK-8法检测K562/A02细胞增殖活性,Giemsa和Hochest 33258染色法观察细胞形态学的变化,流式细胞术检测K562/A02细胞凋亡情况,酶显色活性分析法检测caspase 3酶活性的变化。结果表明,AAVC-Ⅰ能够抑制K562/A02细胞的体外增殖,呈时间和剂量依赖性,48 h时半数抑制浓度为30.988μg/ml。AAVC-Ⅰ作用K562/A02细胞48 h后,镜下可见胞核固缩及凋亡小体的形成。流式细胞术分析显示,随着作用药物的浓度由0增高到50μg/ml,细胞凋亡率也由0.88%升高到53.66%。相比对照组,各实验组caspase 3凋亡蛋白的表达呈浓度依赖性增加(P<0.05)。结论:AAVC-Ⅰ能够有效地抑制人红白血病K562/A02细胞增殖并促进其凋亡,其机制可能与caspase 3蛋白的高表达促进K562/A02细胞凋亡有关。

To investigate the effects of component Ⅰ from Agkistrodon acutus venom（AAVC-Ⅰ） on the biological features of chronic myeloid leukemia cells,K562/A02 leukemia cells were cultured in the presence of AAVC-Ⅰ（6.25-100 μg/ml） and the proliferation status was assayed by CCK-8 method.Morphological changes were observed by inversed microscope after Giemsa and Hochest 33258 staining,and cell apoptosis was detected by flow cytometry.Caspase 3 activity was tested by using Chromogenic Activity Assay Kit.The results showed that AAVC-Ⅰ inhibited the growth of K562/A02 cells in time-and concentration-dependant manners,and the IC50 at 48 h was 30.988 μg/ml.Giemsa and Hochest 33258 staining showed the typical apoptotic features in K562/A02 cells after induction with AAVC-Ⅰ for 48 h.Flow cytometric analysis revealed that the percentage of the apoptotic cells reached from 0.88% up to 53.66% as the treated concentration was elevated from 0 to 50 μg/ml.Compared with the control group,the expression of caspase 3 in the tested group was enhanced in a dose-dependent manner（P0.05）.It is concluded that AAVC-Ⅰ can effectively inhibit the growth and promote apoptosis of K562/A02 cells.Elevated expression of caspase-3 may be attributed to the apoptosis of K562/A02 cells.