马钱子碱对人慢性粒细胞白血病KCL-22细胞的增殖及凋亡作用

目的 探索马钱子碱对人慢性粒细胞白血病KCL-22细胞的增殖抑制及诱导凋亡作用,为白血病的治疗提供新的方向。方法 体外培养白血病KCL-22细胞株,采用MTS法检测不同浓度马钱子碱对KCL-22细胞增殖的影响; 通过FITC-Annexin V/PI双染法检测不同浓度马钱子碱处理后细胞的凋亡作用; 运用Western blotting法检测凋亡相关蛋白的表达水平变化。结果 马钱子碱可明显抑制KCL-22细胞增殖,且具有浓度和时间依赖性。马钱子碱可诱导KCL-22细胞发生凋亡,并以早期凋亡为主,在50～400 μg/ml范围内呈剂量效应关系。经马钱子碱处理后,Bcl-2蛋白表达降低、Bax和Cyt-C蛋白表达增高,Caspase-9,Caspase-3均被切割活化。结论 马钱子碱可显著抑制慢性白血病KCL-22细胞增殖,并可能通过调控Bax/Bcl-2平衡、释放Cyt-C及激活Caspase-3,9依赖的内源性线粒体途径诱导细胞凋亡。

Effects of Brucine on Proliferation and Apoptosis of Chronic Myelogenous Leukemia KCL-22 Cell Line

Objective To study the inhibition and apoptosisof brucine on chronic myelogenous leukemia KCL-22 cells,and then provide a new direction for the treatment of CML.Methods Different concentrations of brucine was added into the KCL-22 cell line in vitro.The inhibition effect of brucine on growth of KCL-22 cells was measured by MTS method.FITC-Annexin-V/PI double labeling method was used to assay the apoptosis rateof KCL-22 after different concentration of brucine treatment.The expression ofapoptosis-related protein was assayed with Western blotting.Results Brucine displayed significant anti-proliferative effect on KCL-22 cells in a dose-and time-dependent manner.Brucine could significantly induced KCL-22 cell apoptosis in the concentration range from 50 μg/ml to 400 μg/ml,mainly for early apoptosis.Western blotting showed that the expression ofBcl-2 was reduced,the expression of Bax and Cyt-C were increased,and the Caspase-9, Caspase-3 were activated.Conclusion Brucinecould inhibits the proliferation of KCL-22 cells,and induce apoptosis possiblythough endogenous mitochondrial pathway by regulating the Bax/Bcl-2 balance,releasing Cyt-C and activating Caspase-9 and Caspase-3.