雄黄_____雄黄；阿霉素；K562(论著)

促凋亡蛋白Caspase3与白血病细胞耐药的相关性

http://www.100md.com 《西安交通大学学报》 2005年第5期

雄黄,,雄黄；阿霉素；K562,AO2细胞株；Caspase3；细胞凋亡；多药耐药,1材料与方法,2结果,3讨论,参考文献

     摘要：目的观察阿霉素(ADR)耐药的人慢性粒细胞白血病细胞株K562/AO2在ADR和雄黄作用下细胞活力、凋亡及加入Caspase3抑制剂前后Caspase3酶活性的变化，探讨促凋亡蛋白Caspase3与白血病细胞耐药的关系。方法 ①四甲基偶氮唑盐法(MTT)测定药物作用下细胞株的细胞活力(A值)；②DNA琼脂糖凝胶电泳观察凋亡细胞的DNA断裂情况；③酶显色活性分析法(CAA)测定Caspase3活性水平的变化。结果 ①MTT法：K562/AO2 细胞的A值与雄黄呈时间、剂量依赖关系。②雄黄作用组有凋亡发生。③未加抑制剂组Caspase3活性明显升高(P<0.05)，对雄黄呈时间、剂量依赖关系，加入抑制剂后此关系消失，对ADR无此关系。结论 Caspase3酶活性水平的变化能显著影响药物诱导肿瘤细胞凋亡的能力，在肿瘤耐药的发生发展中起着重要的作用。

    关键词：雄黄；阿霉素；K562/AO2细胞株；Caspase3；细胞凋亡；多药耐药

    Correlation between drug resistance of leukemic cells and Caspase3Zhao Xinhan, Tian Honggang, Quan Ping, Liu Shaanxi

    (Department of Oncology, First Hospital of Xian Jiaotong University, Xian 710061, China)ABSTRACT: Objective To explore the relationship between drug resistance of leukemic cells and Caspase3, this study took adriamycin (ADR)resistant human chronic granulocytic leukemic cell strain K562/AO2 as research subject, observing the cell survival and the morphological change of cell apoptosis under the action of ADR and arsenic sulfide and the Caspase3 activity before and after putting in the Caspase3 inhibitor. Methods ① The 3(4,5dimethylthiazo2yl)2, 5diphenyltetrazolium bromide (MTT) method was used to determine the cell survival(A value) of K562/AO2 cell strain under the action of ADR and arsenic sulfide. ② DNA agarose gel electrophoresis was performed to observe the DNA cleavage of apoptotic cells. ③ The enzyme colorimetric activity assay (CAA) method was used to measure the change of the Caspase3 activity of K562/AO2 cell strain. Results ① The A value of K562/AO2 cells had a time and dosage dependent relation with arsenic sulfide. ② Apoptosis occurred in the K562/AO2 cell strain affected by arsenic sulfide. ③ Compared with the cell strains with the Caspase3 inhibitor added, the Caspase3 activity of those without the Caspase3 inhibitor increased remarkably (P<0.05). For the K562/AO2 cell strain without the Caspase3 inhibitor added, the activity of Caspase3 had a time and dosage dependent relation with arsenic sulfide; the cell strain with the inhibitor added did not have any time and dosage dependent relation with arsenic sulfide. For the K562/AO2 cell strain, the activity of Caspase3 did not have any time and dosage dependent relation with ADR no matter the Caspase3 inhibitor was added or not. Conclusion Its found that the change of the Caspase3 protein expression level is able to significantly change the ability of drugs inducing cell apoptosis, and it plays an important role in the occurrence and development of drugresistance of tumors. ......

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