PUVA诱导HL-60细胞凋亡时对Fas、FasL表达的影响

Effects of PUVA on apoptosis and expression of Fas and FasL in HL-60 leukemia cells

关键词:  PUVA  白血病  HL-60细胞  凋亡  Fas  FasL

Key words:  PUVA therapy  leukemia  HL-60 cells  apoptosis  Fas  FasL

摘要:

［目的］探讨补骨脂素（PSO）加长波紫外线（UVA）光化学疗法（PUVA）诱导人白血病细胞HL-60凋亡时对Fas、FasL表达的影响。 ［方法］以HL-60细胞为研究对象，以细胞凋亡率、电镜下细胞超微结构改变以及细胞Fas、FasL在基因、蛋白水平的表达为检测指标，观察中药补骨脂中提取的PSO加波长为360 nm的UVA 对人白血病细胞诱导凋亡的作用以及部分作用途径，并采用多因素方差分析法进行统计学处理。 ［结果］（1）PSO、UVA照射及PUVA均可诱导HL-60细胞发生凋亡，对照组、80 μg/mL PSO组、5 min UVA组及PUVA（80 μg/mL PSO+5 min UVA）组凋亡率分别为（10.60±0.31）%、（26.94±0.26）%、（28.53±0.05）%、（37.72±0.09）%，各组间 P＜0.01，PUVA的作用明显强于前两者。（2）电镜下观察经PUVA处理后的HL-60细胞超微结构，可见明显的凋亡形态学改变。（3）PSO、UVA照射及PUVA均可上调HL－60细胞Fas在基因、蛋白水平的表达，下调FasL在基因、蛋白水平的表达，上述四组Fas mRNA的表达量分别为（8.40±0.59）×10³、（1.62±0.36）×10⁵、（1.50±0.26）×10⁵、（7.88±0.38）×10⁶ 拷贝数/μg；Fas蛋白的表达量分别为（3.29±0.39）%、（18.73±1.22）%、（18.17±1.07）%、（47.03±1.36）%；FasL mRNA的表达量分别为（8.69±0.44）×10⁷、（2.71±0.29）×10⁷、（2.64±0.31）×10⁷、（2.61±0.33）×10⁵ 拷贝数/μg；FasL蛋白的表达量分别为（58.67±1.75）%、（37.40±1.73）%、（33.90±1.90）%、（15.60±1.42）%，各组间 P＜0.01，PUVA的作用明显强于前两者。［结论］（1）PUVA可诱导白血病细胞HL-60发生凋亡，作用强于PSO及UVA照射。（2）PUVA诱导HL-60凋亡的途径之一为上调HL－60细胞Fas mRNA表达水平，下调FasL mRNA表达水平。

Abstract:

［Objective］To explore the effects of psoralen (PSO)and long wave ultraviolet A(PUVA) on apoptosis and expression of Fas and FasL in HL-60 leukemia cells.［Methods］The HL-60 cells were taken as the study objects and their apoptosis ratios,ultrastructure changes and the expression of Fas and FasL were detected in order to observe the effects of PSO extracted from Chinese medicine and UVA of wave length 360 nm on human leukemia cells. The factorial design and analysis of variance were used to analyze the interaction among the factors.［Results］(1)PSO, UVA and PUVA all induced the apoptosis and the apoptosis ratios of control group, 80 μg/mL PSO group, 5 min UVA group and PUVA(80 μg/mL PSO+5 min UVA) group were （10.60±0.31）%,（26.94±0.26）%,（28.53±0.05）% and（37.72±0.09）% respectively. The P value among the four groups were 0 (P＜0.01) and the effects of PUVA were stronger than the other two. (2) After HL-60 cells had been treated with PUVA, they all showed obvious ultrastructure change about apoptosis observed under the electron microscope.3.PSO, UVA and PUVA all increased the expression of Fas mRNA and protein and decreased the expression of FasL mRNA and protein. The expression of Fas mRNA of the four groups were （8.40±0.59）×10³,（1.62±0.36）×10⁵,（1.50±0.26）×10⁵ and（7.88±0.38）×10⁶ bp/μg respectively. The expression of Fas protein of the four groups were （3.29±0.39）%,（18.73±1.22）%,（18.17±1.07）% and（47.03±1.36）% respectively. The expression of FasL mRNA of the four groups were （8.69±0.44）×10⁷,（2.71±0.29）×10⁷,（2.64±0.31）×10⁷ and（2.61±0.33）×10⁵ bp/μg respectively. The expression of FasL protein of the four groups were （58.67±1.75）%,（37.40±1.73）%,（33.90±1.90）% and（15.60±1.42）% respectively. The P value among the four groups were 0 (P＜0.01) and the effects of PUVA were stronger than the other two. ［Conclusions］PUVA can induce the apoptosis of HL-60 cells and the effects are the strongest. （2） One of the pathway of PUVA to induce apoptosis is to upregulate the expression of Fas mRNA and downregulate the expression of FasL mRNA.