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

发布时间：2018-03-18 16:42

本文选题：急性白血病　切入点：HL-　出处：《实用医学杂志》2017年03期 　论文类型：期刊论文

【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(P0.01)。干扰LSD1后,HL-60细胞和SHI-1细胞凋亡率明显增加(P0.01),细胞周期阻滞于G_0/G_1期(P0.01)。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。
[Abstract]:Objective: to observe the effect of lentivirus vector mediated shRNA targeting histidine specific demethylase 1 (LSD1) gene on apoptosis and cell cycle of human acute myeloid leukemia cells. Methods: human acute myeloid leukemia cells were constructed with lentivirus vector. Stable cell lines interfering with LSD1 gene in HL-60 cells of promyelocytic leukemia and SHI-1 cells of acute monocytic leukemia. Empty carrier control group and LSD1-shRNA interference group, Real-time fluorescence quantitative PCR and Western blot were used to detect mRNA and protein expression level of LSD1. Annexin V-PE-7-AAD staining was used to detect cell cycle. The expression level of LSD1 mRNA and protein was significantly lower than that of blank control group and empty vector control group. The apoptosis rate of HL-60 cells and SHI-1 cells increased significantly after interfering with LSD1, and the cell cycle was blocked in GStud / G1 phase P0.01.Conclusion the down-regulation of LSD1 expression by LSD1 shRNA is urgent. Apoptosis of myeloid leukemia cells increased and cell cycle G _ 0 / G _ (1) arrest occurred.
【作者单位】：广州医科大学附属广州市第一人民医院血液内科;
【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)
【分类号】：R733.71

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