DZNep诱导HL-60细胞凋亡和生长抑制

3-Deazaneplanocin A induces apoptosis and inhibits cell growth in human acute myeloid leukemia HL-60 cells

[目的]研究S-腺苷同型半胱氨酸水解酶抑制剂DZNep(3-Deazaneplanocin A)对人早幼粒白血病细胞HL-60增殖的影响。[方法]细胞计数法分析DZNep(10~500 nmol/L)处理对HL-60细胞增殖的影响;以DNA片段化、Annexin-V/PI法和线粒体膜电位法检测DZNep处理对HL-60细胞凋亡的影响。Western Blot检测细胞凋亡相关基因Bax和细胞色素C的表达。[结果]50 nmol/L DZNep对HL-60细胞的24、48、72h增殖抑制率达18%、44.5%、81.7%。Annexin-V/PI法显示DZNep能诱导HL-60细胞发生剂量依赖性的细胞凋亡。进一步研究发现,DZNep(50nmol/L)处理24 h诱导23.8%的HL-60细胞发生线粒体途径介导的细胞凋亡,表现为细胞DNA片段化、线粒体膜电位下降和Bax基因表达上调,同时伴有细胞浆内细胞色素C增加。[结论]DZNep能在较低浓度下抑制HL-60细胞增殖并通过激发线粒体凋亡途径诱导HL-60细胞凋亡,具有抗人早幼粒细胞白血病治疗的潜在应用前景。

[Objective]To study the effects of S-adenosyl-homocysteine hydrolase inhibitor DZNep（ 3-Deazaneplanocin A） on HL-60 human promyelocytic leukemia cells growth and apoptosis. [Methods]Cell counting was used to analyze the effects of DZNep（ 10-500 nmol/L） treatment on HL-60 cells growth. Annexin-V/PI double staining,DNA fragmentation,mitochondrial membrane potential analysis and Western Blot assay were used to identify apoptosis in HL-60 cells. [Results]24,48,72 hours treatment of 50 nmol/L DZNep respectively inhibited 18%,44. 5%,81. 7% of HL-60 cells proliferation in a time-and dose-dependent manner. DZnep could induce apoptosis in HL-60 cells. DZnep（ 50 nmol/L） induced 23. 8% of HL-60 cells apoptosis through mitochondria pathway which were identified by DNA fragmentation,decreased mitochondrial membrane potential,increased Bax expression and release of cytochrome C from mitochondria to cytoplasm. [Conclusion]These data suggest that DZNep can inhibit cell growth and induce mitochondria-mediated apoptosis at quit low concentration in HL-60 cells,which has potential value in future treatment of human acute promylocytic leukemia.