采用定量蛋白质组学技术筛选急性髓系白血病HL-60细胞的甲基化相关基因(英文)

Screening for methylation-silenced genes in acute myeloid leukemia HL-60 cell line by a quantitative proteomic approach

目的:筛选人急性髓系白血病细胞株HL-60甲基化沉默基因,为揭示白血病的发病机制及防治提供科学依据。方法:应用荧光差异双向凝胶电泳(two-dimensional fluorescence difference gel electro-phoresis,F-2D-DIGE)分离甲基转移酶抑制剂5-杂氮-2-脱氧胞苷(5-aza-2-dC)处理与未处理的HL-60细胞的总蛋白质,Decyder和PDquest图像分析软件识别差异表达蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术鉴定差异表达蛋白质。结果:建立了5-aza-2-dC处理与未处理的HL-60细胞蛋白质的F-2D-DIGE图谱,图像分析识别了53个差异表达的蛋白质点,质谱分析鉴定了35个差异表达的蛋白质,其中32个蛋白质在5-aza-2-dC处理后的HL-60细胞中表达上调,3个蛋白质表达下调。结论:35个差异表达蛋白可能与甲基化相关,为白血病的表观遗传学研究提供了有价值的信息。

Objective To screen for new methylation association genes in HL-60 to reveal the pathogenesis of leukemia,and provide important theoretical and scientific basis for the prevention and cure of leukemia.Methods Two-dimensional fluorescence difference gel electrophoresis（F-2D-DIGE） was performed to separate the total proteins from acute myelogenous leukemia（AML） cell line HL-60 cells with or without 5-aza-2-deoxycytidine（5-aza-2-dC） treatment.Imaging software Decyder 6.5 and PDQuest were used to detect the differential expression protein spots,and matrix-assisted laser desorption/ionizaion time-of-flight mas spectrometer（MALDI-TOF MS） was adopted to identify the differential expression proteins.Results F-2D-DIGE maps of 5-aza-2-dC-untreated HL-60 and-treated HL-60 cells were established.A total of 53 differential protein spots were detected,and 35 differential proteins were successfully identified.Of the identified proteins,32 proteins were up-regulated,and 3 proteins were down-regulated in HL-60 cells after 5-aza-2-dC treatment.Conclusion Thirty-five differential proteins may be associated with methylation in HL-60 cell line,which provides the important clues for epigenetic study of leukemia.