酞菁锌介导的光动力学疗法对骨髓净化的实验研究  

Effect of zinc phthalocyanine-mediated photodynamic therapy on bone marrow purging, an experimental study

作　　者：黄慧芳[1] 陈元仲[1] 吴勇[1] 

机构地区：[1]福建省血液病研究所

出　　处：《中华医学杂志》2003年第11期986-991,共6页National Medical Journal of China

基　　金：福建省科技项目资助(99 Z 153)

摘　　要：目的　研究酞菁锌光敏剂 (ZnPcS2 P2 )介导的光动力学疗法 (PDT)对模拟慢性粒细胞白血病缓解骨髓的净化作用。方法　采用荧光分光光度法检测K562细胞及正常细胞内的光敏剂含量。锥虫蓝拒染法、MTT比色法、克隆形成实验检测不同浓度ZnPcS2 P2 介导的PDT对K562细胞增殖能力的影响。正常混合集落形成细胞 (CFU Mix)、粒 单核系造血祖细胞 (CFU GM )、红系祖细胞(CFU E)等集落形成实验检测ZnPcS2 P2 PDT对正常造血祖细胞的影响。巢式PCR方法检测酞菁锌介导的PDT作用前后混有不同比例K562细胞的骨髓细胞的bcr ablmRNA表达。结果　 (1 )与ZnPcS2 P2 共孵育 5h ,K562细胞与骨髓正常单个核细胞 (MNC)内的ZnPcS2 P2 含量分别为 1 0 1、2 2(ng/ 5× 1 0 5细胞 ) ,二者比值达到最高值。 (2 ) 0 2 5μg/mlZnPcS2 P2 孵育 5h后 ,用 670nm激光以 53mW/cm2 的功率密度、2 1J/cm2 的能量密度照射对K562细胞集落形成的抑制率为 91 1 % ,而对正常CFU Mix、CFU GM、CFU E等集落形成的抑制率分别为 1 8 0 %、1 8 6 %、1 7 8%。 (3) 0 2 5μg/mlZnPcS2 P2 介导的PDT可完全杀灭按 1∶1 0 0至 1∶1 0 0 0比例混入骨髓正常MNC中的K562细胞。结论　ZnPcS2 P2 介导的PDT能选择性杀伤K562细胞 ,有希望成为新的高效而简便的慢?Objective To probe into the purging effects of zinc phthalocyanine mediated photodynamic therapy (PDT) on simulated remission bone marrow grafts of chronic granulocytic leukemia Methods (1) K562 cells, aline chronic granulocytic leukemia cells, and normal mononuclear cells (MNC) were cultured Zinc phthalocyanine (ZnPcS 2P 2), a photosensitizer, with the terminal concentration of 1 0μg/ml was added into the cultures The K562 cells and normal MNCs in the suspensions were broken Fluoresence spectrophotometry was used to determine the concentration of zinc phthalocyanine in cells at different time points so as to find the optimal time for photodynamic purging process (2) Suspensions of K562 cells and MNCs were made and incubated with zinc phthalocyanine of different concentrations (0 062 5, 0 125, 0 25, 0 5, and 1 0 μg/ml) for 5 hours A blank control group (sodium chloride of the same volume was added), a PDT control group (without photosensitizer), and a photosensitizer control group (zinc phthalocyanine was added without PDT) were established Then the suspensions were irradiated with 670 nm laser Trypan blue dye exclusion technique was used to calculate the number of live cells for a period of 5 days The proliferative potency of K562 cells was detected by MTT colorimetric assay The OD value was detected with ELISA apparatus to calculate the inhibition rate Colony formation of K562 cells and MNCs was determined (3) K562 cells were mixed into normal MNCs at the ratios of 1∶100 and 1∶1 000 so as to create the model of simulated remission bone marrow After PDT treatment, colony formation test was done and nested PCR was used to detect the bcr abl mRNA expression in K562 cells Colony formation test was made on the MNCs treated with PDT The antiproliferative effects of PDT on normal hematopoietic progenitors were evaluated by CFU Mix, CFU GM and CFU E assays Results (1) The zinc phthalocyanine content in the MNCs reached its peak within the first hour of incubat

关 键 词：光动力学疗法 骨髓净化 实验研究 酞菁锌光敏剂 慢性粒细胞白血病 荧光分光光度法 骨髓细胞 

分 类 号：R733.7[医药卫生—肿瘤]