Experimental Studies Of The Effects Of ZnPcH₁-Based-Photodynamic Therapy On SHI-1 Cells

[Objective]①To probe into the kinetic change of ZnPcH1 in normal bone marrow MNC and human monocytic leukemia cell line SHI-1.②To probe into the effects of ZnPcH1 Based Photodynamic Therapy on proliferation and apoptosis influence of SHI-1 cells.[Methods]①Kinetic change of ZnPcH1 in normal bone marrow MNC and SHI-1 cells:Fluorescence intensity of cell extracts was measured by fluoresence spectrophotometry.②The antiproliferative effects of ZnPcH1-PDT on SHI-1 cells: The proliferative potency of SHI-1 cells was detected by MTT colorimetric assay, Typan blue dye exclusion assay and colony formation test. There were three control groups (without any treatment, light only, ZnPcH1 only).③Several parameters were performed to detect the apoptotic cells, i.e. AO/EB stains, TUNEL stain, Cell Cycle test and Annexin-V/PI double staining analysis detected by flow cytometry.④the subcellular localization of ZnPcH1 in SHI-1 cells was determined by laser scanning confocal microscope. [Results]①There existed significant difference of kinetic change of the content ofZnPcH1 in normal hematopoietic cells and SHI-1 cells. After five hours, incubation, the ratio of ZnPcH1 content in SHI-1 cells to that in MNC reached its peak. Therefore, the light delivery was carried out five hours after the incubation.②ZnPcH1-PDT could significantly kill SHI-1 cells in a dose dependent manner. At the concentration of 1.0μM, the inhibitory rate of ZnPcH1-PDT on the colony formation was 100% for SHI-1 cells. However, no substantial differences existed among the three control groups.③ZnPcH1-PDT could induce apoptosis in SHI-1 cells in a time-dependent manner. The phenomena of apoptosis were observed in ZnPcH1-PDT-cells by AO/EB stains. ZnPc-PDT-cells presented nuclear chromatin condensation. TUNEL assay and Annexin-Ⅴstaining show apoptosis rate increase in a time-dependent manner. sub-G1 apoptotic peak can be observed in all ZnPc-PDT-cells by cell cycly analysis.④Detection of the subcellular location of ZnPcH1 was carried out by the fluorescent probe and the results were analyzed by direct observation, false color fusion and coefficient correlation theory. which showed ZnPcH1 located in mitochondria, lysosome and endoplasmic reticulum in SHI-1 cell.[Conclusions]①There exists significant difference of kinetic change of the content of ZnPcH1 in normal hematopoietic cells and SHI-1 cells, After five hours, incubation, the ratio of ZnPcH1 content in SHI-1 cells to that in MNC reached its peak.②ZnPcH1-based photodynamic treatment could inhibit proliferation and induce apoptosis in SHI-1 cells.③ZnPcH1-PDT could induce subcellular multi-sites damages.