【摘要】 目的:观察慢病毒载体介导shRNA靶向沉默组蛋白赖氨酸特异性去甲基化酶1（LSD1）基因对人急性髓系白血病细胞凋亡与细胞周期的影响。方法:利用慢病毒载体构建人急性早幼粒细胞白血病HL-60细胞和急性单核细胞白血病SHI-1细胞LSD1基因干扰的稳定细胞系。设置空白对照组、空载体对照组和LSD1-shRNA干扰组,采用实时荧光定量PCR和Western blot方法分别检测LSD1的mRNA和蛋白表达水平;Annexin V-PE/7-AAD染色后利用流式细胞术检测细胞凋亡;PI染色后检测细胞周期。结果:HL-60细胞和SHI-1细胞LSD1-shRNA干扰组LSD1 mRNA和蛋白表达水平相较于空白对照组及空载体对照组均显著下调（P<0.01）。干扰LSD1后,HL-60细胞和SHI-1细胞凋亡率明显增加（P<0.01）,细胞周期阻滞于G₀/G₁期（P<0.01）。结论:shRNA下调LSD1表达使人急性髓系白血病细胞凋亡增加并发生细胞周期G₀/G₁期阻滞。更多还原

【Abstract】 Objective To investigate the effect of down-regulation of lysine specific demethylase 1(LSD1)by shRNA on the apoptosis and cell cycle of human acute myelogenous leukemia cells.MethodsThe lentiviral vector-mediated LSD1-shRNA was transfected into human acute promyelocytic leukemia HL-60 cells and acute monocytic leukemia SHI-1 cells.The expressions of LSD1 mRNA and protein were examined by real time quantitative PCR and Western blot,respectively.The flow cytometry was applied to detect the apoptosis and cell cycle distribution after Annexin V-PE/7-AAD and PI dying,respectively.ResultsThe expressions of LSD1 mRNA and protein in HL-60 and SHI-1 LSD1-sh RNA group were significantly decreased compared with the blank control group and the negative shRNA group(P<0.01,respectively).The apoptosis levels of HL-60 and SHI-1 cells were significantly increased after knockdown of LSD1(P<0.01).Moreover,the cell cycle distribution in the G0/G1 phases was also significantly increased(P<0.01).Conclusion LSD1-shRNA promotes cell apoptosis and increases the percentage of cells in the G0/G1 phases of human acute myelogenous leukemia cells.更多还原