补骨脂乙素诱导伊马替尼敏感和耐药的慢性粒细胞白血病细胞凋亡

Apoptosis of imatinib-sensitive and imatinib-resistant chronic myelocytic leukemia cells induced by isobavachalcone

目的探讨补骨脂乙素对伊马替尼敏感和耐药慢性粒细胞白血病细胞的影响。方法体外培养人慢性粒细胞白血病伊马替尼敏感细胞株K562s和伊马替尼耐药细胞株K562r,以不同浓度(0、5、10、20、40μmol/L)补骨脂乙素(IBC)处理K562s和K562r细胞不同时间(0、24、48、72 h),锥虫蓝拒染法检测IBC对K562s和K562r细胞活性的影响,Annexin V/PI、Rh123/PI双染后流式细胞术检测细胞凋亡及线粒体跨膜电位的变化,Western blotting检测凋亡相关蛋白的表达。结果锥虫蓝拒染法检测结果显示:IBC对K562s和K562r均有增殖抑制作用,且呈时间-剂量依赖性。流式细胞术检测结果表明,经[BC处理的K562s和K562r出现明显时间-剂量依赖性的凋亡及线粒体膜电位的降低。Western blotting检测结果显示:经IBC处理的K562s和K562r细胞发生caspase-3活化和PARP-1的剪切。结论 IBC能够诱导K562s和K5621,细胞凋亡,线粒体跨膜电位下降可能参与了这一过程。

Objective To explore the effects of isobavachalcone （IBC） on imatinib-sensitive and imadnib- resistant chronic myelocydc leukemia cells. Methods Imatinib-sensitive chronic myelocytic leukemia cells K562s and imatinib-resistant chronic myelocytic leukemia cells K562r were cultured in vitro. K562s and K562r cells were treated by IBC of different concentrations （0, 5, 10, 20, and 40 vmol/L） for different period of time （0, 24, 48, and 72 h）. The effects of IBC on the viability of K562s and K562r cells were detected by the trypan blue exclusion assay. The variations of cell apoptosis and mitochondrial membrane potential were detected by the Annexin V/PI staining and Rh123/PI staining, respectively. The expressions of proteins relevant to apoptosis were detected by the Western blotting. Results The results of trypan blue exclusion assay showed that IBC had an inhibitory effect on the proliferation of K562s and K562r ceils and the effect was time and dose dependent. The results of flow cytometry indicated that the apoptosis and decrease of mitoehondrial membrane potential of K562s and K562r ceils treated by IBC were significantly time and dose dependent. The results of Western blotting showed that IBC induced activation of caspase-3 and cleavage of PARP-1 in K562s and K562r cells. Conclusion IBC can induce the apoptosis of K562s and K562r cells and the decrease of mitochondrial transmembrane potential may involve in this process.