shRNA下调LSD1基因表达对人急性髓系白血病细胞凋亡与细胞周期的影响  

Effect of LSD1 knock-down by small hairpin RNA on the apoptosis and cell cycle of human acute myelogenous leukemia cells

作　　者：林秀梅[1] 曾丽华[1] 许世林[1] 王顺清[1] 毛平[1] 

机构地区：[1]广州医科大学附属广州市第一人民医院血液内科,510180

出　　处：《实用医学杂志》2017年第3期354-358,共5页The Journal of Practical Medicine

基　　金：广东省医学科研基金项目(编号:A2012492);广东省自然科学基金项目(编号:2014A030313676);广州市医药卫生科技项目重点项目(编号:20121A021005);广州市医药卫生科技项目一般引导项目(编号:20141A0100010);广东省科技计划项目(编号:2011B031800290)

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G_0/G_1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G_0/G_1期阻滞。Objective To investigate the effect of down-regulation of lysine specific demethylase 1 （LSDI） by shRNA on the apoptosis and cell cycle of human acute myelogenous leukemia cells. Methods The lcntiviral vector-mediated LSDI-shRNA was transfected into human acute promyelocytic leukemia HL-60 cells and acute monocytic leukemia SHI- 1 cells. The expressions of LSD 1 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 AnnexinV-PE/7-AAD and PI dying, respectively. Results The expressions of LSD 1 mRNA and protein in HL-60 and SHI-1 LSDI-shRNA 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 LSDI-shRNA promotes cell apoptosis and increases the percentage of cells in the G0/G1 phases of human acute myelogenous leukemia cells.

关 键 词：急性白血病 HL-60 SHI-1 赖氨酸特异性去甲基化酶1 细胞凋亡 细胞周期 

分 类 号：R733.71[医药卫生—肿瘤]