光化学疗法对人白血病细胞凋亡及Fas表达的影响

Effects of PUVA on Apoptosis and Expression of Fas in Leukemia Cells

目的观察光化学疗法(PUVA)诱导人白血病细胞HL-60、K562、NB4凋亡时对Fas表达的影响。方法人白血病细胞分别与不同浓度补骨脂素(PSO)、接受或不接受长波紫外线(UVA)照射后共同培养,电镜下观察细胞超微结构改变,荧光定量PCR技术检测细胞Fas基因的表达,流式细胞仪(FCM)检测细胞凋亡率和Fas蛋白的表达,采用多因素方差分析法进行统计学处理。结果PUVA处理后的人白血病细胞超微结构出现明显的凋亡形态学改变;PSO、UVA照射及PUVA可使细胞凋亡率增加,可上调白血病细胞Fas在基因、蛋白水平的表达,PUVA的作用显著强于前两者(P<0.01)。结论PUVA可诱导人白血病细胞发生凋亡,作用强于PSO及UVA照射。PUVA诱导白血病细胞凋亡的途径之一为上调HL-60细胞Fas基因表达。

Objective To explore the effects of psoralen （PSO） and long wave ultraviolet A （PUVA） on apoptosis and expression of Fas in HL-60, K562 and NB4 leukemia cells. Methods The cells were incubated with PSO in different concentrations irradiated with or without UVA. The changes of ultrastructure of cells were observed under the electron microscope. The expression of Fas gene was detected by fluorescent quantitation PCR. The apoptosis ratio and the expression of Fas protein were detected through the flow cytometry. The factorial design and analysis of variance were used to analyze the interaction among the factors. Results There were obvious ultrastructure changes about apoptosis in leukemia cells after treated with PUVA. PSO, UVA and PUVA all increased the apoptosis ratio and expression of Fas gene and protein, and the effects of PUVA were stronger than the other two （P〈0.01）. Conclusions PUVA can induce the apoptosis of leukemia cells and the effects are the strongest. One of the pathways of PUVA to induce apoptosis is to upregulating the expression of Fas gene.