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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。
Objective Caspase-3 and Caspase-8 are abnormally expressed in many kinds of tumors and are closely related to the efficacy of leukemia. This study was designed to investigate the effect of soluble CD40 ligand (sCD40L) on the expression of Caspase-3 and Caspase-8 in human leukemia HL-60 cells and to explore its mechanism of action on HL-60 cells. Methods HL-60 cells were treated with 2.0, 4.0 and 6.0 mg / L sCD40L for 48h. The apoptosis and cell cycle of HL-60 cells were detected by flow cytometry. Caspase-3 and Caspase- 8 gene in HL-60 cells were detected by Western blotting. The protein expression of Caspase-3 and Caspase-8 in HL-60 cells was detected by Western blotting. Results HL-60 cells were treated with sCD40L at concentrations of 2.0, 4.0 and 6.0 mg / L for 48 h, the apoptotic rates of HL-60 cells were (48.17 ± 3.22)% and (60.73 ± 5.46)%, respectively (71.26 ± 5.83)%, which was significantly higher than that of the control group (36.42 ± 2.75)%, F = 36.373, P = 0.003. The expression of Caspase-3 and Caspase-8 mRNA was up-regulated by sCD40L in HL-60 cells in G0 / G1 phase. RT-PCR results showed that the expression of Caspase-3 and Caspase-8 was up-regulated by Western blotting. Conclusions sCD40L can promote the apoptosis of HL-60 cells in a concentration-dependent manner in vitro and block the cells in G0 / G1 phase. The possible mechanism is through activation of Caspase-3 and Caspase-8.