阿糖胞苷联合光动力疗法对人白血病HL-60细胞作用的研究

Combined effects of cytarabine and PDT treatment on leukemia cell HL-60

目的 探讨阿糖胞苷（Ara-c）在癌光啉（PSD-007）介导的光动力疗法（PDT）杀伤人早幼粒白血病HL-60细胞中的联合作用.方法 实验分为空白对照组、PDT单独作用组（PDT l～4组,为光敏剂剂量（5、7.5μg/ml）和激光能量密度（1.2、2.4 J/cm^2）的两两组合）、Ara-c单独作用组（Ara-c A组和Ara-c B组中Ara-c质量浓度分别为0.3 μg/ml和1.2μg/ml）和联合作用组即上述PDT单独作用组和Ara-c单独作用组的两两组合.所有分组分别按3种时序即P24A时序（PDT作用24h后再加入Ara-c共同作用24h）、A24P时序（Ara-c作用24h后进行PDT再共同作用24h）和PA24时序（PDT作用的同时加入Ara-c再共同作用24h）进行处理.采用CCK-8法检测各组细胞活性,用金氏公式分析联合效应,并用流式细胞术检测细胞周期变化.结果 小剂量PSD-007介导的PDT和Ara-c联合时,3种时序的联合作用均表现为协同效应;而大剂量PSD-007介导的PDT和Ara-c联合时,P24A和A24P 2种时序的联合作用效应为协同或相加,PA24时序则主要为相加或拮抗.流式细胞仪检测细胞周期变化结果显示,Ara-c和PSD介导的PDT均能引起细胞周期G0/G1期阻滞.结论 Ara-c与PSD-007介导的PDT联合作用对HL-60细胞的杀伤有协同作用,其协同作用与剂量和作用时序相关,小剂量比大剂量协同效果明显,且Ara-c和PDT间隔24h作用比2者同时作用于该细胞的协同效果明显.

Objective To evaluate the combined effects of photodynamic therapy （PDT） with PSD-007 and cytarabine （Ara-c） on human acute promyelocyte leukemia cell HL-60.Methods The experiments were divided into four groups：control group,PDT-only groups （PDT 1-4 groups：the combination of PSD-007 concentrations （5 μg/ml and 7.5 μg/ml） and the energy density of laser （EDL） （1.2 J/cm2 and 2.4 J/cm^2）,Ara-c-only groups （Ara-c A group：0.3μg/ml,Ara-c B group：1.2μg/ml） and combination groups （the pair-wise combinations of the PDT doses and Ara-c doses above）.All combination groups were treated with three treating methods,including P24A （the cells were treated with PDT for 24 h,and then cocultured with Ara-c for another 24 h）,A24P （treated with Ara-c for 24 h before PDT,and then cocultured with PDT for 24 h）,and PA24 （treated with the Ara-c and PDT for 24 h）.CCK-8 method was used to test the cell viability,and the combined effect was analyzed using King formula.The changes of cell cycles were analyzed using flow cytometry.Results In the case of low-dose PDT,the combination groups showed coordinated effect with all three methods,whereas in the case of high-dose PDT,they showed additive or coordinated effect in P24A and A24P,but it showed antagonistic effect in the schedule of PA24.Cell cycles were inhibited to G0/G1 phase by both PDT and Ara-c.Conclusions Coordinated effects could be found when HL-60 cells were treated with the combination of Ara-c and PDT.The effects depended on the dose of Ara-c and PDT and the operation schedules.The effects at low dose were more obvious than that of high dose,and allowing a 24 h period in between the addition of PDT and Ara-c also promoted the effects.