Résumé
Aim: To investigate the effect of proteasome inhibitor MG132 on the proliferation and apoptosis of acute myeloid leukemia cells. Methods: qRT-PCR was used to detect the expression of ADRM1 mRNA in nine blood tumor cell lines. The expression of ADRM1 in HL60 cells was interfered by shRNA; HL60 cells before and after ADRM1 interfered were treated with different MG132 concentrations for 24 h; Then, the cell proliferation and viability were measured with CCK-8 by microplate reader. Meanwhile, the expressions of ADRM1 and UCH37 protein were detected by Western blot. Apoptosis of HL60 and NB4 cells treated with different MG132 concentrations was analyzed by flow cytometry. Results: ADRM1 mRNA was up-regulated in blood tumor cell lines. ADRM1 shRNA and scrambled shRNA HL60 cells were successfully constructed. Cell proliferation and viability were inhibited by AD-RM1 shRNA interference or decreased with the increase of MG132 concentration; meanwhile, ADRM1 and UCH37 protein expressions were down-regulated. The apoptosis of HL60 and NB4 cells increased with the increase of MG132 concentrations. The apoptotic effect of MG132 on HL60 cells was stronger than that of NB4 cells. Conclusions: ADRM1 mRNA is overexpressed in blood tumor cell lines; ADRM1 down-regulation induces UCH37 protein decrease and cell proliferation inhibition. MG132 induces AML cell apoptosis and restrains the proliferation and viability through down-regulating the expression of ADRM1 and UCH37 protein. The apoptotic effect of MG132 on different types of AML cells exists individual differences.
Résumé
Objective To investigate the effect and the mechanism of different G-CSF-priming protocols on leukemia cell lines (HL-60 and U937) in vitro and provide the clinical guidance to clinical treatment of acute leukemia.Methods The leukemia cell lines HL-60 and U937 were used as model to detect the effects of three drugs alone and combined two drugs (HA) or three drugs (HAG) respectively.Cell viability and cell growth inhibition were performed by cell count kit-8 (CCK-8) assay.Apoptotic marker AnnexinV/PI,cell membrane surface antigen CD11b,cell cycle,mitochondrial membrane potential (JC-1) and Caspase-3 were determined by flow cytometry.Results After using of HAG for 48 h,HL-60 and U937 cells counts were decreased significantly and the apoptotic marker Annexin V was significantly increased. To compare the single drug group with two drug combination group,the result was significantly different (P ＜0.05),and the apoptosis of U937 cells was higher than HL-60 cell line.CD11b expression among the three groups did not change (P ＞ 0.05).Using of CAG and MAG,the mitochondrial nembrane potential of HL-60 and U937cells was increased,the three-drug combination group was significantly higher than single-drug group and control group (P ＜0.05); Caspase-3 was activated,the fluorescence intensities of Caspase-3 of the three-drug combination group and single drug group were significantly higher (P ＜0.05) comparing with the control group.Conclusion HAG regimen could induce leukemia cells to apoptosis through the reduction of mitochondrial membrane potential and the activation of Caspase-3 to induce apoptosis of leukemia cells.