| Background and objectives:Acute promyelocytic leukemia (APL) is characterized by abundant malignant promyelocyticblasts in the bone marrow and severe life threatening bleeding tendency. The genetic hallmark ofAPL is the balanced reciprocal translocation t (15; 17) (q22; q12), resulting in rearrangement ofthe retinoic acid receptor alpha gene (RARa) located onchromosome 17 and the promyelocyticgene (PML) on chromosome 15. APL is a highly curable subtype of acute myeloid leukemia withthe application of all-trans retinoic acid and arsenic trioxide, but there were also some severeadverse effects such as retinoic acid syndrome and toxicity of arsenic trioxide. So it is urgent tofind some secure drugs to cure the APL.Agaricus blazei murrill was an edible mushroom,which was effective to cure liver cancer,colon carcinoma and prostatic carcinoma. NF-κB was a kind of eukaryotic cell nucleartransportation factor, which was distributed widely and participated in many kinds of biologyaction, such as cellular signal transmission, regulating the expression of genes. The abnormalactivation of NF-κB could promote cell growth and restrain apoptosis. There was abnormalactivation of NF-κB in 93% of APL haemopoietic stem cell. In our study, HL-60 cells werecultured in vitro, treated with different concentrations of ABM crude polysaccharides, and thenthe proliferation, apoptosis, and the activation of NF-κB of HL-60 cells were detected.Methods:1. HL-60 cells were cultured in RPMI-1640 medium which contains 10% FBS, 100 U/mLpenicillin and 100 U/mL streptomycin.2. HL-60 cells were treated with ABM crude polysaccharides (1 mg/mL, 2 mg/mL, 5 mg/mL,10 mg/mL) for 12, 24 and 48 hours. The apoptosis and proliferation of HL-60 cells weremeasured by flow cytometry and WST-1 respectively.The activity of NF-κB was measured by immunofluoresce.Results:1. After treated with ABM crude polysaccharides (1 mg/mL, 2 mg/mL, 5 mg/mL, 10 mg/mL )for 12, 24 and 48 hours the proliferation of HL-60 cells were measured by WST-1.①Theproliferation of HL-60 cells was inhibited obviously in 10 mg/mL group after treated withABM crude polysaccharides for 12 h (P<0.05);②The proliferation of HL-60 cells wasinhibited obviously between 2 mg/mL and 10 mg/mL groups after treated with ABM crudepolysaccharides for 24 h (P<0.05);③The proliferation of HL-60 cells was inhibitedobviously in dose dependent manner after treated with ABM crude polysaccharides for 48 h(P<0.05); It was indicated that the proliferation of HL-60 cells was inhibited in dosedependent manner after treated with ABM crude polysaccharides.2. After treated with ABM crude polysaccharides (1 mg/mL, 2 mg/mL, 5 mg/mL, 10 mg/mL )for 12, 24 and 48 hours the apoptotic rate of HL-60 cells was measured by FCM, and the earlyapoptotic rate of HL-60 cells increased in each group comparing with its control (P<0.05).①The early apoptosis of HL-60 cells increased in the 10 mg/mL group more than others aftertreated with ABM crude polysaccharides for 12 h (P<0.05);②The early apoptosis of HL-60cells increased in dose dependent manner after treated with ABM crude polysaccharides for 24h (P<0.05);③After treated for 48 h, the early apoptosis of HL-60 cells increased in the 1-5mg/mL group in dose dependent manner (P<0.05), but the early apoptosis of HL-60 cellsdecreased in the 10 mg/mL group.3. HL-60 cells were treated with ABM crude polysaccharides (1 mg/mL, 2 mg/mL, 5 mg/mL, 10mg/mL ) for 48 h and then measured by immunofluorescence with IPP 6.0 software, the IODof NF-κB in the HL-60 nucleus were 5481.82±5340.31, 4570.21±2828.79, 2320.33±3896.60,1404.11±1615.22, while the IOD of NF-κB in the nucleus of the control group was6952.58±19628.98, It was indicated that the IOD of NF-κB decreased in dose dependentmanner (P<0.05), the NF-κB activity was inhibited by ABM crude polysaccharides. Conclusion:1. ABM crude polysaccharides could inhibit the proliferation of HL-60 cells in a dosedependent manner.2. ABM crude polysaccharides could induce the apoptosis of HL-60 cells in a dose dependentmanner.3. ABM crude polysaccharides could inhibit the activity of NF-κB. |
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