【摘要】 目的 Caspase-3和Caspase-8在多种肿瘤中异常表达,与白血病的疗效等密切相关。本研究分析可溶性CD40配体(soluble CD40ligand,sCD40L)对人白血病HL-60细胞Caspase-3及Caspase-8表达的影响,探讨其对HL-60细胞的作用机制。方法 2.0、4.0和6.0mg/L的sCD40L作用HL-60细胞48h,采用流式细胞术检测HL-60细胞凋亡水平及细胞周期,RT-PCR检测HL-60细胞中Caspase-3和Caspase-8基因的表达水平,蛋白质免疫印迹法检测HL-60细胞中Caspase-3和Caspase-8的蛋白表达。结果 2.0、4.0和6.0mg/L的sCD40L作用HL-60细胞48h后,浓度依赖性诱导HL-60细胞的凋亡,凋亡率分别为(48.17±3.22)%、(60.73±5.46)%及(71.26±5.83)%,与对照组(36.42±2.75)%比较,差异有统计学意义,F=36.373,P=0.003。sCD40L阻滞HL-60细胞在G0/G1期,RT-PCR结果显示,Caspase-3和Caspase-8 mRNA的表达上调;蛋白质印迹法结果显示,Caspase-3和Caspase-8蛋白表达增强。结论 sCD40L在体外能够以浓度依赖性促进HL-60细胞凋亡,阻滞细胞在G0/G1期,其可能机制是通过激活Caspase-3和Caspase-8。更多还原

【Abstract】 OBJECTIVE Caspase-3and Caspase-8are abnormally expressed in a variety of malignant tumors,and closely related to the therapeutic effect of leukemia.The objective of this study was to investigate the effect of soluble CD40 Ligand the expressions of Caspase-3and Caspase-8in leukemia HL-60 cells and to explore its mechanism.METHODS After treatment with sCD40 Lfor 48h,Flow cytometry was used to detect apoptosis and distribution of cell cycle.Real time PCR analysis showed the mRNA expression level of Caspase-3and Caspase-8,Western-blot assay was used to detect the protein level of them in leukemia HL-60 cells.RESULTS After treatment with 2.0,4.0and 6.0 mg/L sCD40 Lfor 48h,the apoptosis of HL-60 cells was induced in concentration-dependent way,the rate was(48.17±3.22)%,(60.73±5.46)% and(71.26±5.83)%,compared with the control group(36.42±2.75)%,and there was statistical significance(F=36.373,P=0.003).The cell cycle of HL-60 was arrested in G0/G1 phase,Real time PCR and Western-blot showed that the mRNA and protein expressions levels of Caspase-3and Caspase-8 were increased.CONCLUSION sCD40 Lcan significantly induce apoptosis of HL-60 cells in a concentration-dependent manner,the cell cycle of HL-60 was arrested in G0/G1 phase,it may be related to their activation of Caspase-3and Caspase-8expression.更多还原