CAJ | 학술논문

目的探讨小檗碱对耐药急性淋巴细胞白血病细胞株EU-4的凋亡诱导作用及其机制.方法 0、10、1 00 μmol／L小檗碱作用EU-4细胞72 h,采用流式细胞术检测细胞凋亡,免疫印迹法检测Caspase-3、PARP及X连锁凋亡抑制蛋白(XIAP)的蛋白表达,比色法检测Caspase-3的活性变化,基因沉默技术观察降低XIAP表达对于细胞凋亡的影响.结果 0、10、100μmol／L小檗碱作用EU-4细胞72 h后凋亡率分别为(9.08±1.20)％、(22.36±2.16)％、(59.81±4.17)％,呈剂量-效应关系.凋亡过程中Caspase-3的活性增强,分别为1.70±0.25、1.92±0.10、2.89±0.25.小檗碱抑制EU-4细胞XIAP的表达(P＜0.05),并呈剂量和时间依赖关系.单纯下调XIAP蛋白的表达引起细胞凋亡,对照siRNA转染组和XIAP siRNA转染组细胞凋亡率分别为(9.23±1.66)％和(22.15±0.63)％.结论小檗碱可以诱导急性淋巴细胞白血病EU-4细胞凋亡,XIAP下调引起的Caspase-3的激活参与了EU-4细胞的凋亡.
목적 탐토소벽감대내약급성림파세포백혈병세포주EU-4적조망유도작용급기궤제.방법 0、10、1 00 μmol／L소벽감작용EU-4세포72 h,채용류식세포술검측세포조망,면역인적법검측Caspase-3、PARP급X련쇄조망억제단백(XIAP)적단백표체,비색법검측Caspase-3적활성변화,기인침묵기술관찰강저XIAP표체대우세포조망적영향.결과 0、10、100μmol／L소벽감작용EU-4세포72 h후조망솔분별위(9.08±1.20)％、(22.36±2.16)％、(59.81±4.17)％,정제량-효응관계.조망과정중Caspase-3적활성증강,분별위1.70±0.25、1.92±0.10、2.89±0.25.소벽감억제EU-4세포XIAP적표체(P＜0.05),병정제량화시간의뢰관계.단순하조XIAP단백적표체인기세포조망,대조siRNA전염조화XIAP siRNA전염조세포조망솔분별위(9.23±1.66)％화(22.15±0.63)％.결론 소벽감가이유도급성림파세포백혈병EU-4세포조망,XIAP하조인기적Caspase-3적격활삼여료EU-4세포적조망.
Objective To study the molecular mechanism of berberine induced apoptosis in chemoresistant EU-4 acute lymphocytic leukemia cells.Methods EU-4 cells were treated with 0,10 and 100 μmol/L berberine for 72 h.The apoptosis induced by berberine was detected by flow cytometry.The expression of Caspase-3,PARP and X-linked inhibitor of apoptosis protein (XIAP) were determined by Western bolt assay.Caspase-3 activity was measured using microplate reader.After transfected with XIAP siRNA,the apoptosis was detected by flow cytometry.Results After treated with 0,10 and 100 μmol/L berberine for 72 h,the apoptosis rates of EU-4 cells were (9.08±1.20) ％,(22.36±2.16) ％ and (59.81±4.17) ％,respectively.Berberine induced potent apoptosis in a dose-dependent manner.The apoptosis involved activation of Caspase-3.The Caspase-3 activities were 1.70±0.25,1.92±0.10 and 2.89±0.25,respectively.Berberine inhibited XIAP expression in a dose-and time-dependent manner (P ＜ 0.05).Down-regulation of XIAP by siRNA increased apoptosis of EU-4 cells.The apoptosis rates were (9.23±1.66) ％ and (22.15±0.63) ％.Conclusion Berberine could induce apoptosis of EU-4 cells,and inhibition of XIAP leading to Caspase-3 activation is responsible for the apoptotic effect on EU-4 cells.