Study On BMK-induced Apoptosis In Human Acute Myeloid Leukemic HL-60 Cell

Study on BMK-induced Apoptosis in Human Acute Myeloid Leukemic HL-60 cell Objectives: To investigate the effect of Buthus martensii Karsh scorpion venom (BMK) on cell growth and apoptosis in human acute promyelocytic leukemic HL-60 cell and its anti-tumor molecular mechanism.Methods:(1) Detecting the effect of BMK on cell growth in HL-60 cell line with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay;(2) Examining the potential role of BMK-induced apoptosis in HL-60 cell by flow cytometry (FCM) analysis with Annexin V-FITC Apoptosis Detection Kit and caspase-3 staining kit;(3) Determining the wild p53 gene expression level affected by BMK in HL-60 cell via Reverses transcription polymerase chain reaction method.Results:(1) We examined the effect of BMK on cell growth with MTT assay and found that BMK had a dose-dependent inhibitory effect on HL-60 cell growth after 48h treatment. The 50% inhibitory concentration (IC50) of BMK for HL-60 cells was 139.67mg/L.(2) Flow cytometric analysis demonstrated that after incubation with BMK100μg/ml and 200μg/ml for 24h, there was a dose-dependent increase in Annexin V binding to HL-60 cells, 13.95 % and 26.53 % respectively, compared to untreated cells 6.89 % (P<0.05).(3) Also, FCM analysis showed the caspase-3 activation by BMK in HL–60 cells, with an increased amount (7.59) % of active caspase3-positive cells comparing that (2.82) % of untreated control group.(4) We did not see p53 specific band by Reverses transcription polymerase chain reaction (RT-PCR), indicating that BMK-induced apoptosis in HL–60 cells is p53 non-dependent.Conclusions:(1) BMK has a dose-dependent growth inhibitory effect in human acute myeloid leukemic HL-60 cells.(2) BMK induces apoptosis of HL-60 cells.(3) BMK activates caspase 3 in HL-60 cells.(4) The induction apoptosis by BMK in HL–60 cells is p53 non-dependent.