基于二氢卟吩e6的光动力疗法对骨髓净化作用的实验研究

Huang Hui-fang, Chen Yuan-zhong, Wu Yong

Fujian Institute of Hematology, Union Hospital, Fujian Medical University, Fuzhou 350001, China.

Chin Med J (Engl). 2005 Jan 20;118(2):105-10.

BACKGROUND

An effective purging technique plays an important role in autologous hematopoietic stem cells transplantation. Photodynamic therapy (PDT) provides a novel approach for this purpose. This study dealt with the purging effects of di-sulfo-di-phthalimidomethyl phthalolcyanine zinc (ZnPcS2P2)-based photodynamic therapy (ZnPc-PDT).

METHODS

Fluorescence intensity of cell extracts was measured using a fluorescence spectrophotometry. The proliferative potency of K562 cells and HL60 cells was detected using MTT colorimetric assay, Typan blue dye exclusion method, colony formation test. The proliferative potency of normal hematopoietic cells was evaluated using mixture colony-forming unit (CFU-Mix), granulocyte-macrophage colony-forming unit (CFU-GM), and erythrocyte colony-forming unit (CFU-E) assays. K562 cells were mixed with normal mononuclear cells (MNCs) at ratios of 1:100 and 1:1000 for creating the model of simulated remission bone marrow. Colony formation test and nested-RT-PCR were carried out to detect the residual K562 cells in cell mixture.

RESULTS

After a 5-hour incubation with ZnPcS2P2, the content of ZnPcS2P2 in normal MNCs was the lowest value. At the same time, the content in K562 cells and HL60 cells was very high. Therefore, the time point was selected as the optimal one for irradiating the cell suspensions. ZnPc-PDT could significantly kill proliferative K562 cells and HL60 cells in a dose-dependent manner. At the concentration of 1.0 microg/ml, the inhibitory rate of ZnPc-PDT on the colony formation was 100% for K562 cells, 89.7% for HL60 cells. 0.25 microg/ml ZnPc-PDT could completely photoinactivate residual K562 cells in the simulated remission bone marrow. Under an identical condition, the inhibitory rates of CFU-Mix, CFU-GM, CFU-E were 18.0%, 18.6%, and 17.8% respectively.

CONCLUSION

ZnPc-PDT appears to be a promising approach for bone marrow purging.

背景

有效的清除技术在自体造血干细胞移植中起着重要作用。光动力疗法（PDT）为此提供了一种新方法。本研究探讨了基于二磺基二邻苯二甲酰亚胺甲基酞菁锌（ZnPcS2P2）的光动力疗法（ZnPc-PDT）的清除效果。

方法

采用荧光分光光度法测定细胞提取物的荧光强度。采用MTT比色法、台盼蓝染料排斥法、集落形成试验检测K562细胞和HL60细胞的增殖能力。采用混合集落形成单位（CFU-Mix）、粒细胞-巨噬细胞集落形成单位（CFU-GM）和红细胞集落形成单位（CFU-E）试验评估正常造血细胞的增殖能力。将K562细胞与正常单核细胞（MNCs）按1:100和1:1000的比例混合，建立模拟缓解骨髓模型。采用集落形成试验和巢式RT-PCR检测细胞混合物中残留的K562细胞。

结果

与ZnPcS2P2孵育5小时后，正常MNCs中ZnPcS2P2的含量为最低值。同时，K562细胞和HL60细胞中的含量非常高。因此，选择该时间点作为照射细胞悬液的最佳时间点。ZnPc-PDT能以剂量依赖的方式显著杀伤增殖的K562细胞和HL60细胞。在浓度为1.0μg/ml时，ZnPc-PDT对K562细胞集落形成的抑制率为100%，对HL60细胞的抑制率为89.7%。0.25μg/ml ZnPc-PDT可使模拟缓解骨髓中残留的K562细胞完全光失活。在相同条件下，CFU-Mix、CFU-GM、CFU-E的抑制率分别为18.0%、18.6%和17.8%。