神经前体细胞表达下调因子8类泛素修饰抑制因子MLN4924对单核巨噬细胞白血病细胞增殖和凋亡的影响

发布时间：2018-09-03 10:41

【摘要】：目的观察神经前体细胞表达下调因子8(NEDD8)类泛素修饰抑制因子MLN4924对单核巨噬细胞白血病细胞增殖的影响,并探讨其作用机制。方法 MLN4924 0.25~4.0μmol·L-1与U937细胞、THP-1细胞、Mo7e细胞和RAW264.7细胞作用24 h,采用MTT法检测细胞存活。MLN4924 0.25~8.0μmol·L-1与RAW264.7细胞作用24 h,用Western蛋白质印迹法检测NEDD8类泛素化修饰的主要底物cullin蛋白表达水平。MLN4924 0.5μmol·L-1作用于RAW264.7细胞24 h,用流式细胞仪检测细胞周期和细胞凋亡率。结果 MLN4924 0.25~4.0μmol·L-1作用24 h对RAW264.7细胞、U937细胞、THP-1细胞和Mo7e细胞增殖具有抑制作用,IC50分别为0.52,3.11,2.89和0.76μmol·L-1;对正常小鼠骨髓巨噬细胞增殖无明显影响。MLN4924 0.25~8.0μmol·L-1作用24 h,NEDD8类泛素化修饰的cullin蛋白表达减少(P0.01),表明RAW264.7细胞NEDD8修饰逐渐减少。MLN4924 0.5,1.0和2.0μmol·L-1作用于RAW264.7细胞24 h,G0/G1期细胞百分率由正常对照组的(61.3±1.3)%分别升高到(64.5±1.5)%(P0.05),(69.5±2.3)%(P0.01)和(72.8±1.7)%(P0.01),细胞凋亡率由正常对照组的(1.5±0.3)%分别升高到(2.3±0.5)%,(4.3±0.9)%(P0.05)和(7.5±0.8)%(P0.01)。结论 MLN4924能够明显抑制单核巨噬细胞白血病细胞增殖,并诱导其细胞周期阻滞和细胞凋亡。
[Abstract]:Objective To observe the effect of MLN4924 on the proliferation of monocyte-macrophage leukemia cells and explore its mechanism. Methods MLN4924 0.25-4.0 micromol.L-1 and U937 cells, THP-1 cells, Mo7e cells and RAW264.7 cells were treated for 24 hours. MTT assay was used to detect cell survival. The expression of Cullin protein in RAW264.7 cells treated with 924 0.25-8.0 micromol.L-1 for 24 hours was detected by Western blotting. The expression of Cullin protein in RAW264.7 cells treated with MLN4924 0.5 micromol.L-1 for 24 hours was detected by flow cytometry. Results The expression of Cullin protein in RAW264.7 cells treated with MLN4924 0.25-4.0 micromol.0 micromol.L-1 for 24 hours was detected by Western blotting. The proliferation of W264.7 cells, U937 cells, THP-1 cells and Mo7e cells was inhibited by IC50 of 0.52, 3.11, 2.89 and 0.76 micromol L-1, respectively. No significant effect on the proliferation of bone marrow macrophages was observed in normal mice. MLN 4924 0.5, 1.0 and 2.0 micromol L-1 treated RAW264.7 cells for 24 hours, the percentage of G0/G1 phase cells increased from (61.3 1.3)% to (64.5 1.5)% (P 0.05), (69.5 2.3)% (P 0.01) and (72.8 1.7)% (P 0.01) respectively, and the percentage of apoptosis increased from (1.5 0.3)% to (2.3 (4.3 0.9)% (P 0.05) in the normal control group. Conclusion MLN4924 can inhibit the proliferation of monocyte-macrophage leukemia cells and induce cell cycle arrest and apoptosis.
【作者单位】：北京中医药大学中药学院;军事医学科学院基础医学研究所免疫学研究室;
【基金】：国家科技重大专项(2013ZX09103003)~~
【分类号】：R329.2

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