KDM6A基因对急性髓系白血病细胞增殖、凋亡的影响  

Effects of KDM6A gene on proliferation and apoptosis of acute myeloid leukemia cells

作　　者：骆婷婷 刘洋[1] 张瑞[1] 于文燕[2] LUO Tingting;LIU Yang;ZHANG Rui;YU Wenyan(Laboratory of Hematology Center,First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China;Basic Medical College of Xinjiang Medical University)

机构地区：[1]新疆医科大学第一附属医院血液病中心实验室,乌鲁木齐830054 [2]新疆医科大学基础医学院

出　　处：《山西医科大学学报》2021年第9期1109-1114,共6页Journal of Shanxi Medical University

基　　金：新疆维吾尔自治区自然科学基金项目(2020D01C237)。

摘　　要：目的体外观察过表达和敲减赖氨酸特异性去甲基化酶6A(KDM6A)基因对急性髓系白血病(AML)细胞增殖、凋亡的影响,为急性髓系白血病基因靶向治疗提供理论依据。方法体外培养U937细胞,分为5组:U937细胞组、sh-control组(转染慢病毒空载体)、sh-KDM6A组(转染shKDM6A慢病毒载体)、pcDNA3.0组(转染pcDNA3.0)、pcDNA3.0-KDM6A组(转染pcDNA3.0-KDM6A过表达载体)。采用MTT法和流式细胞术分别检测细胞增殖和凋亡,荧光定量PCR检测KDM6A、c-Myc、Cyclin D1、Bcl-2、Bax、Caspase-3 mRNA表达水平,Western blot检测KDM6A、H3K27me3、c-Myc、Cyclin D1、Bcl-2、Bax、Caspase-3蛋白表达水平。结果与sh-control组相比,sh-KDM6A组细胞存活率、c-Myc、Cyclin D1、Bcl-2 mRNA及H3K27me3、c-Myc、Cyclin D1、Bcl-2蛋白表达显著升高(P<0.05),凋亡率、KDM6A、Bax、Caspase-3 mRNA及蛋白显著下降(P<0.05);与pcDNA3.0组相比,pcDNA3.0-KDM6A组细胞存活率、c-Myc、Cyclin D1、Bcl-2 mRNA水平及H3K27me3、c-Myc、Cyclin D1、Bcl-2蛋白水平显著降低(P<0.05),凋亡率、KDM6A、Bax、Caspase-3 mRNA水平及蛋白水平显著升高(P<0.05)。结论KDM6A过表达在急性髓系白血病中通过抑制细胞增殖促进细胞凋亡来发挥抗肿瘤作用。Objective To observe the effects of overexpression and knockdown of lysine-specific demethylase 6A(KDM6A)gene on the proliferation and apoptosis of acute myeloid leukemia(AML)cells in vitro,so as to provide theoretical basis for gene targeted therapy of AML.Methods U937 cells were cultured in vitro,and divided into five groups:U937 cell group,sh-control group(transfected with slow virus empty vector),sh-KDM6A group(transfected with shKDM6A lentiviral vector),pcDNA3.0 group(transfected with pcDNA3.0)and pcDNA3.0-KDM6A group(transfected with pcDNA3.0-KDM6A).KDM6A was knocked down by infection with lentivirus in U937 cells,while the expression of KDM6A was up-regulated by transfection with pcDNA3.0-KDM6A overexpression plasmid.MTT assay and flow cytometry were used to detect the cell proliferation and the apoptosis respectively.The mRNA levels of KDM6A,c-Myc,Cyclin D1,Bcl-2,Bax and Caspase-3 were detected by fluorescence quantitative PCR,and the protein expression levels of KDM6A,H3K27me3,c-Myc,Cyclin D1,Bcl-2,Bax and Caspase-3 were detected by Western blot.Results Compared with sh-control group,the cell survival rate,and the expression levels of c-Myc,Cyclin D1,Bcl-2 mRNA and H3K27me3,c-Myc,Cyclin D1,Bcl-2 proteins were significantly increased in sh-KDM6A group(P<0.05),while the apoptosis rate,and the mRNA and protein levels of KDM6A,Bax,Caspase-3 were significantly decreased(P<0.05).Compared with pcDNA3.0 group,the survival rate,and the levels of c-Myc,Cyclin D1,Bcl-2 mRNA and H3K27me3,c-Myc,Cyclin D1,Bcl-2 proteins in pcDNA3.0-KDM6A group were significantly decreased(P<0.05),while the apoptosis rate,and the mRNA and protein levels of KDM6A,Bax,Caspase-3 were significantly increased(P<0.05).Conclusion Overexpression of KDM6A plays an anti-tumor role in AML by inhibiting the cell proliferation and promoting the cell apoptosis.

关 键 词：赖氨酸特异性去甲基化酶6A 细胞增殖 细胞凋亡 急性髓系白血病 

分 类 号：R733.7[医药卫生—肿瘤]