小白菊内酯诱导耐药白血病细胞K562/ADR凋亡的研究  

Apoptosis of leukemia K562/ADR cell with multidrug resistance induced by parthenolide

作　　者：石倩倩[1] 丁亚辉[2] 祁瑞哲 高瀛岱 杨铭 纪庆 罗成[1] 

机构地区：[1]天津科技大学食品工程与生物技术学院,天津300457 [2]南开大学药学院,天津300071 [3]实验血液学国家重点实验室,中国医学科学院血液病医院(血液学研究所),天津300020

出　　处：《中国药理学通报》2012年第12期1736-1740,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 8100137,21072106)

摘　　要：目的初步探讨小白菊内酯(PTL)对耐药白血病细胞K562/ADR细胞增殖和凋亡的影响及其作用机制。方法MTT法及LDH释放法检测小白菊内酯对K562和K562/ADR细胞增殖抑制作用,流式细胞术检测细胞凋亡和细胞内活性氧(ROS)的变化,Western blot法检测凋亡相关蛋白Bax、Bcl-2、caspase-3、caspase-9的表达变化。结果 PTL抑制K562/ADR细胞的增殖,呈剂量依赖,半数抑制浓度(IC50)值分别为(4.36±0.37)μmol.L-1、(10.6±2.81)μmol.L-1。10μmol.L-1PTL作用24 h诱导K562/ADR细胞的凋亡率为(31.21±0.40)%,其IC50值与凋亡率与K562细胞相比差异无显著性(P>0.05)。经10μmol.L-1PTL处理1 h,K562/ADR细胞内ROS增加;处理24 h,Bcl-2与Bax的比值降低、caspase-3、caspase-9酶源蛋白表达降低。用N-乙酰半胱氨酸(NAC)预处理细胞,能抑制细胞ROS水平升高和细胞凋亡。结论 PTL能诱导耐药白血病细胞凋亡,作用机制与其刺激细胞内ROS升高相关。Aim To explore the effects of parthenolide（PTL） on cell cytotoxicity and the apoptosis of K562/ADR,and its mechanisms.Methods MTT assay and LDH assay was applied to detect the effect of inhibition on K562,K562/ADR for 72 h.The apoptosis rate and reactive oxygen species（ROS） level of cells were examined by flow cytometry.The expression of proteins Bax,Bcl-2,Bax,caspase-3 and caspase-9 were measured by western blot.Results PTL was shown to inhibit the growth of K562/ADR in a dose-dependent manner,with mean IC50 of（4.36±0.37） μmol·L-1,and LC50 of（10.6±2.81） μmol·L-1.The apoptosis rate of K562/ADR was（31.21±0.40）% induced by 10 μmol·L-1 PTL for 24 h,which had no significant difference from those of K562 cell.Increase of ROS levels were shown in K562 and K562/ADR cells after treatment of 10 μmol·L-1 PTL for 1 h.The expression of Bcl-2/Bax,caspase-3 and caspase-9 decreased induces by 10 μmol·L-1 PTL for 24 h.ROS increase and cell apoptosis of K562/ADR could be inhibited by the antioxidant N-acetyl-L-cysteine（NAC） pretreatment.Conclusion PTL induces multidrug resistance cell K562/ADR apoptosis,and its possible mechanism is associated with the ROS increases in the cellular induced by PTL.

关 键 词：小白菊内酯 细胞凋亡 活性氧 耐药 白血病 N-乙酰半胱氨酸 

分 类 号：R285.5[医药卫生—中药学]