补骨脂素加长波紫外线对HL60细胞凋亡的影响.DOCVIP

临床医学论文-补骨脂素加长波紫外线对HL60细胞凋亡的影响 【摘要】? 　　目的：探讨补骨脂素（psoralen, PSO）加长波紫外线（ultravioletA, UVA）的光化学疗法（PUVA）对人急性髓细胞白血病（FABM2）细胞株HL60细胞凋亡的作用及部分作用途径。方法：以HL60细胞株为研究对象，以0、10、20、40、80 μg/ml PSO加波长为360 nm的UVA，作用于HL60细胞，通过流式细胞仪检测HL60细胞在PUVA后的凋亡情况；电镜下观察细胞超微结构改变；荧光定量聚合酶链式反应（polymerase chain reaction, PCR）及流式细胞技术检测Fas配体（Fas ligand, FasL)在基因和蛋白水平的表达情况。结果：单纯PSO、单纯UVA照射及PUVA均可诱导HL60细胞发生凋亡，PUVA的作用显著强于前两者。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。PSO、UVA照射及PUVA均可下调FasL在基因、蛋白水平的表达，PUVA的作用显著强于前两者。结论： PUVA可诱导白血病细胞HL60发生凋亡，作用强于PSO及UVA照射。HL60被PUVA诱导凋亡后FasL基因表达水平下调。 【关键词】? 白血病 HL 60细胞 细胞凋亡 Fas配体 紫外线疗法 光化学疗法 补骨脂素 　　Methods: The HL60 cells were taken as the study objects and their apoptosis rates, ultrastructure changes and the expression of FasL were detected in order to observe the effects of PSO and ultravioletA (UVA) of wave length 360 nm. The factorial design and analysis of variance were used to analyze the interaction among the factors. 　　Results: PSO, UVA and PUVA all induced the apoptosis and the effects of PUVA were stronger than those of the other two. After HL60 cells had been treated with PUVA, they all showed obvious ultrastructure changes due to apoptosis observed under the electron microscope. PSO, UVA and PUVA all decreased the expressions of FasL gene and protein. The effects of PUVA were stronger than those of the other two. 　　Conclusions: PUVA can induce the apoptosis of HL60 cells and the effects are stronger than those of PSO or UVA alone. The expression of FasL gene in HL60 cells is downregulated during the apoptosis induced by PUVA. 　　Keywords: leukemia; HL60 cells; apoptosis; Fas ligand; ultraviolet therapy; photochemotherapy; psoralen 　　补骨脂素（psoralen, PSO）是中药补骨脂种子中的主要成分，研究表明其具有光敏性质与抗肿瘤活性。本实验从中药补骨脂中提取出PSO，以不同浓度PSO加波长为360 nm的紫外线（ultravioletA, UVA）（简称PUVA），作用于HL60细胞，通过流式细胞仪检测HL60细胞在PUVA后的凋亡情况；电镜下观察细胞超微结构改变；荧光定量聚合酶链式反应（polymerase chain reaction, PCR）及流式细胞技术检测Fas配体（Fas ligand, FasL)在基因和蛋白水平的表达，以探讨PUVA法诱导HL60细胞凋亡的作用及部分作用途径。 　　1? 材料与方法 　　1.1? 细胞株? 人急性髓细胞白血病M2