5-ALA介导的光动力治疗在耐药白血病中的作用机制研究  

Mechanism Research of ALA-PDT on Resistant Leukemia

作　　者：韩晓凤[1] 黄洪晖[1] 倪蓓文[1] 钟璐[1] 钟济华[1] 陈芳源[1] 

机构地区：[1]上海交通大学医学院附属仁济医院血液科,上海200127

出　　处：《应用激光》2014年第1期76-80,共5页Applied Laser

摘　　要：目的:本研究观察了5-氨基乙酰丙酸介导的光动力治疗(ALA-PDT)对耐药白血病细胞株HL-60/ADR的杀伤作用及对耐药白血病原代细胞存活率的影响。方法:以耐药白血病细胞株HL-60/ADR为实验模型,同时在11例白血病患者原代细胞中进行检测。实验分为4组,对照组、单纯ALA组、单纯光照组及ALA+PDT组。用MTT法测定细胞的存活率,采用阳离子脂质荧光探针JC-1检测线粒体跨膜电位,用Real-time PCR检测PDT前后HL-60/ADR细胞株中bcl-2基因及多药耐药基因MRP的表达变化。结果:ALA-PDT后HL-60/ADR细胞株线粒体跨膜电位出现快速下降,0,2,4h时线粒体跨膜电位崩塌的细胞比例分别升高至55.91%±2.60%、64.27%±1.08%、82.17%±0.43%,与对照组相比皆有显著差异(P<0.05),呈时间依赖性。而单纯ALA组和单纯光照组则无明显变化。HL-60/ADR细胞株在ALA-PDT后Bcl-2和MRP基因均呈明显下降趋势。在初发和复发难治急性白血病原代细胞中ALA+PDT组均显示较强的光动力效应。结论:ALA-PDT诱导的HL-60/ADR细胞的杀伤可能与其影响线粒体跨膜电位有关,即通过影响线粒体功能促进细胞凋亡。同时表明ALA介导的光动力作用部分是通过在基因转录水平下调抗凋亡基因Bcl-2而促进凋亡的发生,另一方面通过下调耐药基因MRP的表达而部分逆转耐药。同时ALA-PDT对原代白血病细胞同样有较大的抑制作用。Objective.. To investigate the mechanisms of AI.A-PDT induced cell death in resistant HI.-60/ADR leukemia cell line and to determine the inhibition of cell proliferation after ALA-PDT on resistant primary leukemia cells. Methods： Using HL 60/ADR ceil line as a model, the ceils were divided into four groups, only ALA group, only PDT group, ALA＋PDT group and control group. Flow cytometry analysis was used to detect variations in Mitochondrial Membrane Potential （MMP） by u- sing the lipophilic cation 5,5＇,6,6＇- tetrachloro- 1,1＇, 3,3- tetraethyl benzimidazol-carbocyanine iodide （JC-1）. Real time-PCR were used to evaluate the mRNA expression of Bcl2 and MRP. Results：The fraction of cells with the disrupted membrane po- tential was significantly higher after ALA combined with irradiation with time dependence. The results revealed that ALA PDT down regulated Bcl2 and MRP expression in HL-60/ADR cell line. ALA-PDT can inhibit the proliferation of leukemia primary cells. Conclusions：ALA-PDT may alter mitochondrial function of the cells, resulting in loss of MMP and down regulated Bcl2 and MRP gene expression.

关 键 词：5-氨基乙酰丙酸 光动力学治疗 HL-60 ADR 细胞凋亡 线粒体 bcl- 2 多药耐药蛋白 

分 类 号：R730.57[医药卫生—肿瘤]