氟达拉滨联合丙戊酸对慢性粒细胞白血病细胞凋亡诱导作用

Synergistic Effect between Fludarabine and Valproic Acid in Apoptotic-inducing of Human K562 Cells in Vitro

目的 研究氟达拉滨（Fludarabine,FDB）联合丙戊酸（valproic acid,VPA）对慢性粒细胞白血病（chronic myeloidleukemia,CML）细胞株K562增殖及凋亡的影响,并探讨其可能的作用机制.方法 慢性粒细胞白血病细胞株K562,单独使用不同浓度的FDB（1,5,10μmol/L）或VPA（1,2.5,5 mmol/L）,或联合使用FDB和VPA处理K562细胞,采用MTT法检测细胞增殖情况,并观察其有效的作用浓度;流式细胞仪检测K562细胞周期变化;western blot检测凋亡相关蛋白及组织蛋白酶B的表达变化.结果 单独使用FDB或VPA均可明显抑制K562细胞的增殖,呈剂量依赖性,并使K562细胞停滞在G0/G1期,联合使用VPA能增强FDB对K562细胞的凋亡诱导作用.同时检测到FDB和VPA能诱导溶酶体中组织蛋白酶B的表达.结论 联合使用FDB和VPA可诱导K562细胞溶酶体中组织蛋白酶B的表达及活性,从而启动溶酶体介导的细胞凋亡过程,明显抑制K562细胞的增殖,并促进K562细胞的凋亡,提高治疗效果,为临床抗肿瘤治疗提供新的指导方向.

Objective To investigate the proliferation inhibitory and apoptotic induction effect of fludarabine （FDB） and valp roic acid （VPA） in human chronic myeloid leukemia （CML） cells K562 cells.Methods The cell proliferation inhibitory rates of K562 cells treated with different concentration of FDB （1,5,10 μmol/L） or VPA （1,2.5,5 mmol/L） alone and in combination were detected by MTT assay.The change of cell cycle of K562 cells was detected by flow cytometer.The ex pression level of apoptosis maker-caspase-3 and cathepsin B was measured by western blot.Results Different concentration of FDB or VPA alone or in combination could obviously inhibit the proliferation of K562 cells in a dose dependent manner （P＜0.05）,FDB and VPA could block K562 cells in G0/G1 phase and activate the cell apoptosis through lysosome cathep sin B pathway.Conclusions FDB and VPA had anti tumor effect on K562 cells in vitro through inhibiting cell proliferation and inducing cell apoptosis,which might be related to the regulation of lysosome cathepsin B pathway.FDB and VPA may be used as new anti-tumor agents for the treatment of CML.