土木香内酯对K562/ADR细胞增殖的抑制作用及其机制探讨

Inhibitory effect of alantolactone on the proliferation of K562/ADR cells and its mechanism

目的 探讨土木香内酯对慢性髓性白血病耐药细胞株K562/ADR增殖的抑制作用及其机制.方法 选用不同浓度（0、1、2、4、6、8、10 μmol/L）土木香内酯处理K562/ADR细胞不同时间,用MTT法测定土木香内酯对K562/ADR细胞增殖的抑制作用,流式细胞术检测土木香内酯对K562/ADR细胞凋亡的影响,Westem blot检测凋亡相关蛋白的表达.结果 土木香内酯对K562/ADR细胞有明显的增殖抑制作用,且呈明显的时间-剂量依赖性,其作用24 h的半数有效抑制浓度（IC50）值约为4.7μtmol/L （P＜0.05）;流式细胞术检测结果显示,0、2.5、5、7.5 μmol/L土木香内酯处理24 h后细胞凋亡率分别为1.35％、16.91％、29.61％和46.26％,亦呈剂量依赖性（P＜0.05）;同时土木香内酯能够降低Bcl-2、BCR-ABL的表达,上调Bax、细胞色素C、活化的caspase-9、caspase-3和PARP的表达.结论 土木香内酯对K562/ADR细胞具有明显的增殖抑制作用,其作用可能是通过caspase依赖的线粒体途径诱导细胞凋亡实现的.

Objective To explore the inhibitory effect of alantolactone on the proliferation of adriamycin-resistant human chronic myelogenous leukemia cell line K562/ADR cells and its mechanism.Methods K562/ADR cells were treated with various concentrations of alantolactone （0,1,2,4,6,8,and 10 μmol/L） for different time points.Cell viability was analyzed with MTT assay.The effect of alantolactone on the apoptosis of K562/ADR cells was measured by flow cytometry.The expression of apoptosis-related proteins after treatment with alantolactone was analyzed using Western blot.Results Alantolactone could effectively inhibit the proliferation of K562/ADR cells in dose-and time-dependent manner,the IC50 value of alantolactone treatment of K562/ADR cells for 24h was 4.7 μmol/L （P〈0.05）.Flow cytometric analysis displayed that the apoptotic rates were 1.35%,16.91%,29.61% and 46.26%,respectively,after treatment with alantolactone at 0,2.5,5 and 7.5 μmol/L.Meanwhile,the expression of Bcl-2 and BCR-ABL proteins were significantly decreased and that of Bax,cytochrome C,cleaved-caspase-9,cleaved-caspase-3 and cleaved-PARP increased by alantolactone treatment.Conclusion Alantolactone had obvious inhibitory effect on the proliferation of K562/ADR cells through the caspase dependent mitochondrial （or intrinsic） apoptotic pathway.