论文部分内容阅读
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。
Objective To study the mechanism of the apoptosis of leukemia cells induced by mannycin, a farnesyltransferase inhibitor. Methods The leukemic cell line U937 and HL-60 cells were treated with 2 μmol / L of mC for different time, and apoptosis was detected by flow cytometry. Western blotting was used to detect the expression of cytochrome C, caspase9, caspase8 and caspase3. Mitochondrial membrane potential (△ ψm) was determined by fluorochrome JC-1 assay, and caspase inhibitor Z-VAD-fmk was used to block the activation of caspase to explore the role of caspase in apoptosis induced by vancomycin. Results The △ ψm of U937 and HL-60 cells treated with 2 μmol / L of moxibustion for 10 h decreased significantly (relative values were 0.51 ± 0.07 and 0.41 ± 0.06, respectively) (P <0.01). The release of cytochrome c from mitochondria was induced by cyclophosphamide in the cytoplasm of the cells. Activation of caspase-9, caspase 8 and caspase-3.50 μmol / L of Z-VAD-fmk completely blocked the activation of caspase, but only partially blocked the induction of cyclophosphamide The apoptosis rate of U937 and HL-60 cells decreased by 51.69% and 56.47%, respectively. CONCLUSION: Fcyomycin induces the apoptosis of U937 and HL-60 cells through the mitochondrial pathway.