Qinba Mushroom (Fa-2-b-β) Exhibit Anticancer Effects Against Chronic Myeloid Leukemia Cells

ObjectiveIn this study,We selected the the extract from Agaricus blazei Murrill-Qbsg Polysaccharide,FA-2-b-βto act on chronic myeloid leukemia K562 cell line and chronic myeloid primary cells.Agaricus Blazei Extract（FA-2-b-?）Induced apoptosis in Chronic myeloid leukemia cells by Wnt/β-catenin、caspase3 signaling.The mechanism of the effect on the apoptosis of chronic myeloid leukemia provides a Provide experimental basis for clinical application.MethodsThe cell proliferation rate was measured by CCK8 method at different time points.Apoptosis was detected by flow cytometry.The chronic myeloid leukemia model was constructed by tail vein injection and subcutaneous inoculation in NOD/SCID mice.To observe the mice to form tumor,random death row pathological model mice,specific antibody markers,Immunofluorescence and immunohistochemi-stry detection,We regularly observed the peripheral white blood cells and hemoglobin counts.ResultsFA-2-b-βinhibited primary CML bone marrow cells proliferation significantly in a concentration dependent manner.In 24h,48h,72h,r_K562-24h=-0.9111、r_K562-48h=-0.9067、r_K562-72h=-0.9213、r_p-24h=-9082、r_p-48h=-9442、r _p-72h=-9546,The total apoptotic K562cell percentages of the control group were1.18 and 0.11%at 24 and 48 h,respectively,and it increased to 93.4%,95.73%and 71.83%and 9.9/25.21 and 79.49/71.83%after treatment with FA-2-b-β（1.2,1.8 and 2.4 mg/mL）,respectively,for 24 and 48 hours,The total apoptotic primary cell percentages of the control group were3.94 and 10.7%at 24 and 48 h,respectively,and it increased to81.13%/79.81%and 17.8/30.4%and49.21/52.89 after treatment with FA-2-b-β（1.2,1.8 and 2.4 mg/mL）,There was an increase in G1 phase,but a partially decrease in the S phase in a dose-dependent manner.G1 and S phases were 89.75 and 0%in the control,but G1 phases were increased to33.73%/37.18%,34.94%/37.01%,37.36%/43.09%and 41.53%/60.64%at24 hours and 33.73%/37.65%,37.01%/45.07%,56.95%/53.11%and 58.99%/60.66%but S phases were partially reduced to 46/46%,54/49%and 58/27%at 24 and 46/46%,48/40%and 37/29%48 hours in the presence of 1.2,1.8 and 2.4 mg/ml FA-2-b-β,then RT-qPCR and Western-blot analysis was performed to assess the level of Wnt/β-catenin-related apoptosis gene and protein expression at 48h.Bcl-2 expression was reduced and Bax expression increased in a dose-dependent manner.FA-2-b-βreduces cyclin D1 expression for binding to the promoter region and reduces theβ-catenin protein and gene expression for nuclear localizationβ-catenin,BCL-2,CD44,CyclinD1,LEF/TCF-4,Cjun,mmp expression was reduced in a dose-dependent manner.Respectively,Experiment group and positive control group has similar treatment effect that significantly prolonged the survival of CML mice model and efficiency of bloods counts,body weights Most remarkedly,Although,White bloods cell counts is still high.Bcl-2,Cytochrome C,caspase8,caspase9gene/protein expression was reduced and Bax,caspase3 expression increased in a dose-dependent manner.ConclusionAnti-cancer activity of proteoglucan was to via inhibiting proliferative and inducting pro-apoptotic procedures.Daily intake of mushrooms containing proteog-lucan is sufficient as an alternative medicine in the management of CML,and might provide useful information for development of a novel therapeutic target in this area.