The Mechanism of Norcantharidin Promoting Apoptosis in Diffuse Large B Lymphoma Cells Through UHRF1
To explore the effects of norcantharidin(NCTD)on apoptosis of diffuse large B-cell lymphoma(DLBCL)cells and its molecular mechanisms.Human DLBCL cell lines SU-DHL-4 and U2932 were used to divided into control group and NCTD group(0,2.5,5,10,20,40 μmol/L).Cell survival rate was detected by MTT method,apoptosis rate was detected by flow cytometry,and Western blot was used to detect the expression of apoptosis related proteins.The differences of UHRF1 in DLBCL and normal tissues were analyzed by TCGA and GEPIA databases,and the correlation between UHRF1 gene and apoptosis was analyzed through Spearman analysis.AutoDock 4.0 software was used to perform macromolecular docking between UHRF1 and NCTD,and Western blot was used to detect the impact of NCTD on the expression of UHRF1 protein.Our study found that compared with the control group,the survival rate of DLBCL cells in the NCTD(10,20,40 μmol/L)group was significantly reduced(P＜0.05),and the trend was concentration and time dependent.The apoptosis rates of the NCTD 10,20,and 40 μmol/L groups were significantly increased(P＜0.05,P＜0.01),while the expression of Cleaved-Caspase-3 protein increased and the level of Bcl-2 protein decreased.The ratios of Cleaved-caspase-3/Caspase-3 and Bax/Bcl-2 were significantly increased(P＜0.01).Through GEPIA and TCGA database analysis,it was found that UHRF1 is highly expressed in DLBCL compared to normal tissues(P＜0.05,P＜0.001),which is a risk factor affecting the survival of DLBCL and closely related to cell apoptosis.Molecular docking results showed that UHRF1 could form a stable conformation with NCTD,and the binding energy was-24.36 kJ/mol.The expression of UHRF1 protein in NCTD group was significantly lower than that in control group(P＜0.05).Therefor,high expression of UHRF1 is an independent risk factor affecting the survival of DLBCL patients,and NCTD can promote DLBCL cell apoptosis by downregulating the expression of UHRF1 protein.
diffuse large B-cell lymphomanorcantharidinapoptosisUHRF1
NETL
NSTL
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