| Objective:To clear IL-24for bone marrow mononuclear cells of leukemia cultured in vitro the influence ofproliferation and apoptosis. Preliminary disscuss the possible mechanisms of IL-24induce BMMNCapoptosis, as to provide theoretical basis for clinical treatment of leukemia.Methods:(1) Collecting bone marrows which confirmed leukemia from May2012to October2013in theHematology Department of the First Auxiliary Hospital of the Shihezi University outpatient andhospitalization were without any cytokines、hormones and chemical drug treated, It was include30cases ofpatients with acute leukemia group including15cases of acute lymphoblastic leukemia,15cases of acutemyeloid leukemia.15cases of normal bone marrow. The separation of mononuclear cells with humanlymphocyte separation medium/density gradient centrifugation and then cultured in vitro. Adjust the celldensity to2x106/ml, at the concentration of0,20,50,100ng/ml IL-24vaccinate cells,each concentrationhad three holes,respectively in0,24,48,72hours collect cells after cultured in vitro.(2) The proliferation ofthese cells were detected by MTT assay and cell count.(3)The apoptosis rate of these cells were detectedby flow cytometry.(4) The expression of bcl-2and caspase-3mRNA of these cells were detected byRT-PCR method.(5) The change of the morphology of cells were observed by Wright stain. The databaseof the obtained data was established in Excel,using spss17.0statistical analysis.Results:（1）MTT assay showed that dosing concentration in0,20,50ng/ml, along with the increase ofthe concentration of IL-24, adult acute leukemia BMMNC proliferation inhibition rate increased, and whenthe concentration of IL-24to100ng/ml, the inhibition rate was lower than50ng/ml. The effect of IL-24was also time-dependent. when in0,24,48h its proliferation inhibition rate was increased with timegradually rise, but when in72h its proliferation inhibition rate was relatively lower than48h. Dosing48hwhen compared to other point in time,different concentrations of IL-24,had the strongest cell proliferationinhibition rates.Normal control group was not affected after dosing,cell count results were consistent withthe above results.（2）Add0,20,50,100ng/ml IL-2448h, the differences of acute leukemia bone marrowmononuclear cells apoptosis rates were statistically significant（P<0.05）.After treated with the same,thedifferences of the normal bone marrow mononuclear cells apoptosis rate were not statisticallysignificant(P>0.05).In dosing concentration of50ng/ml,dosing time for48h,the cell apoptosis rate was thehighest（.3）Dosing concentration0,20,50,100ng/ml IL-24for48h,In the concentration of0、20、50ng/ml，along with the increase of the concentration of IL-24,the expression of bcl-2in acute leukemia wasdecreased,(but in100ng/ml its expression intension was enhanced compared with50ng/ml); In theconcentration of0、20、50ng/ml，along with the increase of the concentration of IL-24the expressions ofcaspase-3in acute leukemia BMMNC were increased,(but in100ng/ml its expression intension wasdecreased). Through the statistical analysis of the expression of acute leukemia in different dosingconcentrations had different ststistically significants,there was no ststistically significant differencecompared with the control cells.(4) Wright’s staining showed the no apoptotic cell nucleus was large,roundand the shape of the early apoptotic cell was irregular,and the nuclear chromatin concentrated,karyopyknosis presented hemispherical、circular or crescent-shaped,stained darker than noapoptosis cells.Conclusion:(1) That IL-24inhibit the proliferation of the adult acute leukemia BMMNC.(2) That IL-24can induce adult leukemia BMMNC apoptosis.(3)That IL-24can decrease the expression of bcl-2mRNAand increase the expression of caspase-3mRNA.(4)The experimental results provide pathogenesis and newtheoretical basic for clinical treatment of adult acute leukemia. |
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