ZnPcS₂P₂-Based-Photodynamic Therapy On Su-DHL-4 Cells And Suberlocalizationon Of Photosensitizer ZnPcS₂P₂ In K562 Cells,HL-60 Cells And Su-DHL-4 Cells

High-dose chemotherapy supported by hematopoietic stem/progenitor cells transplantation (HSCT) presents an effective access of final cure of leukemia.Photodynamic therapy. Autologous HSCT (Auto-HSCT) manifests several advantages over allogenous HSCT (Allo-HSCT), such as easy transplantation of grafts, obviating the need for a HLA-matched donor, without the restriction of age and graft versus host disease (GVHD). Auto-HSCT, therefore, presents a promising approach for leukemia therapy. Auto-HSCT, however, bears the risk of reinfusing residual leukemic cells that contribute to disease relapse and purging procedures, therefore, are needed to eliminate the risk. Myeloablative therapy with bone marrow or peripheral blood progenitor cell transplantation may play an important role in the treatment of lymphoma, A major concern in autografting programs is that cell harvest contains occult tumor cells that may contribute to disease relapse. No report about the effects of PDT on follic lymphoma cells .PDT is a novel cancer therapy that has been shown to destroy many types of solid tumors in animal models and in humans.PDT Typically uses red light to excite photosensitive drugs that preferentially localize within the tumor tissue. The excited photosensitizers can react with molecular oxygen (O₂) to produce singlet oxygen (¹O₂) and other reactive oxygen species that ultimately destroy the tumor. singlet oxygen (¹O₂) has its life-time less than 0.05 s,in cells with a diffuse length of less than 0.02um and the targets of PDT are thus the loci where the photosensitizer is localized.A new amphipathic photosensitizer, the di-sulfonated -phthalimidomethyl phthalocyanine zinc (ZnPcS₂P₂), is a second-generation PDT photosensitizer that is a highly efficient inducer of apoptosis in K562 cells,HL-60 cells and su-DHL-4 cells and appears to activate a mitochondrial pathway of apoptosis on K562 cells,HL-60 cells. Therefore, we were interested in determining the subcellular localization of ZnPcS₂P₂ in these cells. The term''localization''will be defined in this paper as the areas of the cell where the photosensitizer accumulates. One technique used to determine localization is doublelabel confocal fluorescence microscopy. In this technique, a drug and an organelle-specific dye are both administered to a cell. The drug and organelle dye must have distinguishable fluorescence bands so that separate fluorescence images may be acquired of the drug and dye localization within the same cell via double-label confocal microscopy. The fluorescence images of the drug and of the organelle organelle specific dye provide the subcellular distribution of the two compounds. The areas of overlap between the two images provide information about the spatial accumulation of the drug within different organelles, but this is not an accurate method of determining the amount of overlap between two images; subjectivity can reduce the reliability of these descriptions. More accurate information about drug localization can be obtained through quantitative analysis.Thus we using the correlation coefficient. quantifying the localization of the photosensitizer in the organelles of K562 cells,HL-60 cells and su-DHL-4 cells.The major results and conclusion were obtained as follows :1.ZnPcS₂P₂ -PDT could significantly kill su-DHL-4 cells in a dose dependent manner. At the concentration of 0.41μg/ml, the inhibitory rate of ZnPcS₂P₂ -PDT using MTT assays is 50%.2. Apoptosis in su-DHL-4 induced by ZnPcS₂P₂ -PDT. TUNEL assay showed that ZnPcS₂P₂ -PDT induced apoptosis of su-DHL-4 cells in a dose-dependent manner. Staining of cells with AO/EB revealed that ZnPcS₂P₂ -PDT induced nuclear chromatin condensation and fragmentation .3.ZnPcS₂P₂ located on the all the three organelle.this study provided direct evidence that mitochondria,lysosome and endoplasmic reticulum play an important role in the apoptosis in K562 cells ,HL-60 cells and su-DHL-4 cells induced by ZnPcS₂P₂ - PDT. From above results, we conclude that ZnPcS₂P₂-PDT was able to inhibit the proliferation of su-DHL-4 cells and induce su-DHL-4 cells apoptosis. ZnPcS₂P₂ located on mitochondria,lysosome and endoplasmic reticulum.