组蛋白去甲基化酶KDM3B在急性髓系白血病中的靶基因鉴定

Identification of the histone demethylase KDM3B target gene in acute myeloid leukemia

目的:组蛋白H3赖氨酸9位(H3K9)的甲基化是造血和白血病发生中关键的表观遗传学修饰,由组蛋白甲基化酶和去甲基化酶精确调控。组蛋白H3K9去甲基化酶KDM3B则是一个潜在的白血病发生抑制基因。我们旨在探讨白血病中KDM3B的潜在靶基因。方法:干涉多个急性髓系白血病细胞株中的KDM3B基因后进行微阵列芯片实验,采用基因集富集分析(GSEA)及Venn分析数据,筛选出若干KDM3B潜在的靶基因,并用实时定量PCR对CDKN1A基因的结果进行验证。结果:对比GSEA的分子标签数据库中标志基因集(H)和已建立的基因集(C2),在NB-4和PLB-985细胞中(分别影响),与KDM3 B正相关的基因分别有2 375和2 007个,其中P<0.05的基因分别有89和67个;与KDM3B负相关的基因分别有1 882和2 250个,其中P<0.05的基因分别有62和67个,包括FLT3、NRAS、EVI1和IL4等基因。对比GSEA的分子标签数据库中染色体位置(C1)、已建立的基因集(C2)、模序(C3)、肿瘤相关基因集(C4)、GO基因集(C5)、致瘤基因集(C6)、免疫学基因集(C7)和标志基因集(H),在NB-4和PLB-985细胞中(共同影响),与KDM3 B正相关的基因有4 815个,其中P<0.05的基因有130个;与KDM3B负相关的基因有4 400个,其中P<0.05的基因有97个,包括POU5F1和CDKN1A等基因。干扰NB-4和PLB-985细胞中KDM3B基因的表达,我们选取影响最强的600个探针用Venn分析,在两种细胞系中表达均下降的基因有16个,包括血液病相关基因CCDN3、TRIM24、MAPKI3、SOX6等;在NB-4细胞中表达降低但在PLB-985细胞中表达升高的基因有17个,包括RARRES3和CD58等基因。实时定量PCR结果显示干扰PLB-985细胞中KDM3B后CDKN1A的表达是对照组的1.18倍。结论:KDM3B调控急性髓系白血病相关基因,CDKN1A是一个潜在的KDM3B靶基因。

Objective：Histone H3 lysine 9（H3K9） methylation is the key epigenetic modification in hematopoiesis and leukemia,and is regulated by histone methylase and demethylase.Histone H3K9 demethylase KDM3B is a potential leukemia suppressor gene.We aimed to investigate the target gene of KDM3B in leukemia.Methods：Microarray analysis was performed on multiple acute myeloid leukemia cell lines after intervention of KDM3B,and the results were verified by Real-time quantitative PCR.Results：Compared with the gene set hallmark（H） and curate（C2） in the molecular tag database of the GSEA,2 375 and 2 007 genes were positively correlated with KDM3B in NB-4 and PLB-985 cells,among which 89 and 67 were P〈0.05,1 882 and 2 250 negative genes associated with KDM3B,of which 62 and 67 were P〈0.05,including FLT3,NRAS and EVI1,IL4 genes.Compared with the gene set position（C1）,curate（C2）,motif（C3）,computational（C4）,gene ontology（C5）,oncogenic signatures（C6）,immunologic signatures（C7） and hallmark（H）,in the NB-4 cells and PLB-985 cells,there were 4 815 genes were positively correlated with KDM3B,among which there were 130 genes with P〈0.05.4 400 genes were negatively correlated with KDM3B,and 97 genes were P〈0.05,including POU5F1 and CDKN1A and so on.To investigate the expression of gene in NB-4 and PLB cells,we elected 600 of the most influential probes with Venn analysis,and 16 genes were down-regulated in both cell lines,including blood cell related genes CCDN3,TRIM24,MAPKI3,SOX6 and 17 genes reduced expression in NB-4 cells but up-expressed in PLB-985 cells,including RARRES3 and CD58.Real-time quantitative PCR results showed that the expression of CDKN1A was 1.18 times higher than that of normal cells after interfering with KDM3B in PLB-985 cells.Conclusion：KDM3B regulates acute myeloid leukemia-associated genes,and CDKN1A is a potential KDM3B target gene.