【摘要】 目的研究法尼酰基转移酶抑制剂手霉素(Manumycin)诱导白血病细胞凋亡的机制。方法用2 μmol/L 手霉素处理白血病细胞系 U937和 HL-60细胞不同时间,用流式细胞术检测细胞凋亡。免疫印迹技术检测细胞色素 C、caspase9、caspase8、caspase3的表达。用荧光染料 JC-1检测法测定线粒体膜电位(△ψm),并用 caspase 抑制剂 Z-VAD-fmk 阻断 caspase 的活化,探讨 caspase 在手霉素诱导白血病细胞凋亡中的作用。结果 2 μmol/L 手霉素处理 U937和 HL-60细胞16 h,△ψm显著下降,相对值分别是0.51±0.07和0.41±0.06(P<0.01)。手霉素诱导细胞色素 C 从线粒体释放到细胞质,激活 caspase-9、caspase8和 caspase-3。50 μmol/L 的 Z-VAD-fmk 可完全阻断 caspase 激活,但仅部分阻断手霉素诱导的 U937和 HL-60细胞凋亡,细胞凋亡率分别减少51.69%和56.47%。结论手霉素通过线粒体途径诱导 U937和 HL-60细胞凋亡。更多还原

【Abstract】 Objective To investigate the apoptosis induced by manumycin in U937 and HL-60 cell lines,and to explore the role of mitochondria apoptotic pathway in manumycin-inducing apoptosis.Methods Leukemic cells line U937 and HL-60 were treated by manumycin at 2 μmol/L for different time.Apoptosis of leukemia cells was detected by flow cytometry.The cytosolic proteins were extracted using a digitonin buff- er.The protein expression of cytochrome C,caspase-9,caspase-8,and caspase-3 were determined by western blot.Mitochondrial membrane potential was detected by JC-1.Results In U937 and HL-60 cells,manumy- cin induced mitochondrial depolarization after 6 h treatment.The average red/green fluorescence ratios at 6 h were significantly(P<0.01)lower than those at time 0,being 0.51±0.07 and 0.41±0.06 for control group respectively.Manumycin induced cytochrome C release from the mitochondria into the cytosol after 6 h treatment,and activated caspase-9,caspase-8,and caspase-3 after a 16h treatment.The broad-spectrum caspase-inhibitor Z-VAD-fmk at 50 μmol/L was able to inhibit caspase cleavage completely,but only reduced the manumycin-induced apoptosis rates by 51.69% and 56.47% in U937 and HL-60,respectively.Conclu- sion Manumycin induced apoptosis in U937 and HL-60 cell lines via mitochondria apoptotic pathway.更多还原