The Different Proteomics Research Of Arsenic Trioxide Inducting Leukemia Cell Apoptosis

bjective: To observe the effect of apoptosis induction about variousconcentrations of As₂O₃on AML HL-60cell line and the differential proteinexpression before and after the intervention on AML HL-60cell.To exploremechanism of As₂O₃for inducting HL-60cell apoptosis by searching some of thedifferences between the function of the protein,and provide the experimentalbasis for As₂O₃curing AML.Method:1.Using HL-60cell line as leukemiamodel.2.Grouping and intervention:The experiments were divided into sixgroups,including the experimental groups to join with0.1,1,2,4,8umol/L As₂O₃and control group without any drug.With three replicates hole,using the tumorcells in vitro cultivation of72h.3.By Wright staining method for identification ofcell and observation the morphology of rats about As₂O₃on apoptosis of HL-60cell.4. Using Flow Cytome try to test the effects of different concentrations ofAs₂O₃on HL-60cell line5.Useing Nano HPLC and Q Exactive look fordifferences protein..5.1Each group was extracted from total protein after cultruedfor72h,5.2Quantify the protein by BRADFORD, Confirm again by SDS-PAGEelectrophoresis.5.3Enzyme solution in the solution.5.4Mass spectrometry.5.5Bioinformatics analysis.6. Statistical analysis:Statistical anaylsis waspeformedsing by using the statistical programs SPSS16.0, data were expressed as average±standard devidation. P values less than0.05is considered to bestatistical significance. Result:Wright staining demonstrated that cultured cells areleukocytes，with the drug concentration increasing apoptotic cells is more obvious，less cytoplasm,cell size smaller, chromatin to the nuclear membrane close to thenuclear membrane and part of cell membrane with filopodia-like protrusions,intracellular apoptotic bodies appeared and nuclear pyknosis, karyorrhexis the typicalapoptotic morphological changes In an inverted optical microscope,2.Flowcytometric detection results Indicates, the effect of apoptosis Induction about As₂O₃on AML HL-60cell is In a concentration dependent enhancement. Compared withcontrol group,there was significant difference.3. Mass spectrometric identificationand search database results Indicate that there are102protein before and afterintervention of high expression or low expression. Among them,PARP,ATPase andEF-2, etc promote or inhibite the process of As₂O₃inducing HL-60cellatoapoptosis.Lay the foundation of clinical application for arsenic trioxidetreatment for leukemia.