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

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 electrophoresis, 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，为白血病表观遗传学研究提供了有价值的信息。

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

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.