Study Of Apoptosis Bufotalin Induces Of Acute Promyelocytic Leukemia Cells

Objective:Through Bufflin effect on acute leukemia-HL 60 cells,explore the Bufflin of acute leukemia cells-HL 60 induced apoptosis inhibition of proliferation of research,so as to provide basis for the treatment of acute leukemia Bufflin,to provide theoretical basis for clinical applications.Methods:Using different concentrations of Bufflin applyed on the acute myeloid leukemia cell HL-60;(1)cell proliferation assay: apply trypan blue dyeing experiment on application of Bufflin of HL-60 cell staining,cell count,cell proliferation inhibitor cell vitality by formula calculation,draw the proliferation inhibition curve;The application of MTT method to detect cell proliferation and cell vitality,and the cell energy and inhibition rate are calculated by the formula,and the curve is drawn.The application flow cytometer detects the DNA content and the cell cycle in the cell,and tests the DNA content and the period of the cells within the cell.(2)determination of apoptosis: Hochest33258 / PI staining method was applied to different concentrations of toad after his spirit action after cell double dye fluorescence microscope to observe cells form and quantity;Annexin V-FITC/PI double marking method was applied to double dye-HL 60 cells,flow cytometry is used to detect apoptosis percentage,for Bufflin – HL-60 cells for cell proliferation and apoptosis after testing.Results:1.Bufflin can effectively restrain the HL 60 cell proliferation,and drug concentration dependence,through calculating the IC50 of 6.627?g/ml,and can block cells in G0 / G1 phase,thus inhibiting cell proliferation and induce apoptosis.2.By using the Hoechst33258 / PI double dye experiment and the Hoechst33258/PI double labeling method,we found that Bufflin was able to induce acute myeloid leukemia cell HL-60 apoptosis.3.Bufflin 1.25?g/ml concentration cell apoptosis appeared,in Bufflin concentration is 5.0?g/ml ~ 10.0?g/ml can achieve good effect induced apoptosis,and the first part of the experiment on cell proliferation inhibition of determination of the experimental results are consistent.Conclusion:1.This experiment through the detection of cell proliferation ability,apply trypan blue dye from experiment,determined by MTT and flow cytometry instrument to detect cellular DNA content and cell cycle experiment method,revealing the Bufflin of acute myeloid leukemia HL 60 cells with inhibition of proliferation,and presents the drug concentration dependence;2.Through the determination of cell apoptosis experiment,it is concluded that the Bufflin can inhibit the HL-60 cell proliferation activity,induction of apoptosis,and its effect in the HL-60 apoptosis induced by the effect of drug concentration,1.25 ? g/ml drug concentration cell apoptosis appeared,in toad his spirit concentration of 5.0?g/ml-10.0?g/ml can achieve good effect induced apoptosis;3.Bufflin has the effect of inhibiting the proliferation and induced apoptosis of the HL-60 cell line in acute myeloid leukemia.4.This study has clearly Bufflin of HL-60 cells can inhibit proliferation and induce the apoptosis of effect,but the specific mechanism is not yet clear,needs further research.