蛋白质组学技术分析5-aza-2dC处理白血病细胞HL-60前后的差异表达蛋白质  

Proteomic analysis of differentially expressed proteins in leukemia cell HL-60 following treatment with 5-aza-2'-deoxycytidine

作　　者：谭潭[1] 梁婷[1] 汤参娥[1] 肖艳华[1] 易红[1] 张鹏飞[1] 彭芳[1] 陈主初[1] 肖志强[1] 

机构地区：[1]中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室,长沙410008

出　　处：《国际病理科学与临床杂志》2009年第2期93-99,共7页Journal of International Pathology and Clinical Medicine

基　　金：教育部跨世纪优秀人才培养计划基金(教育部科技函[2002]48);湖南省科技厅重点科研项目(06SK2004);湖南省自然科学基金项目(07JJ5031)~~

摘　　要：目的:研究甲基化转移酶抑制剂5-杂氮-2’-脱氧胞苷(5-aza-2’-deoxycitydine,5-aza-2dC)对人急性髓系白血病细胞HL-60细胞蛋白质表达谱的影响,探讨5-aza-2dC抗急性髓系白血病作用的机制。方法:采用二维凝胶电泳(2-DE)技术分离5-aza-2-dC处理与未处理HL-60细胞的总蛋白质,PDquest图像分析软件识别差异蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定差异蛋白质点。然后采用Western印迹检测差异蛋白质半乳糖凝集素1(Galectin-1)在药物处理与未处理HL-60细胞中的表达水平。结果:建立了5-aza-2dC处理与未处理HL-60细胞蛋白质的2-DE图谱,识别了35个差异表达的蛋白质点,鉴定了27个差异表达的蛋白质,其中包括Galectin-1在内的21个蛋白质在5-aza-2dC处理后的HL-60细胞中表达上调,6个蛋白质表达下调或缺失。Western印迹结果证实Galectin-1在5-aza-2dC处理HL-60细胞中表达上调。结论:21个表达上调蛋白质的编码基因可能是HL-60细胞的甲基化失活基因,它们可能与急性髓系白血病发病以及5-aza-2-dC抗人急性髓系白血病细胞作用有关。Objective To investigate the effect of methylation transferase inhibitor 5-aza-2＇ -de- oxycitydine （5-aza-2dC） on the protein expression profile in human acute myeloid leukemia （AML） cell line HL-60, and to explore the mechanism of 5-aza-2dC on AML. Methods Two-dimensional gel electrophoresis （2-DE） was performed to separate the proteins of treated and untreated HL-60 ceils with 5- aza-2dC; PDQuest software was used to analyze 2-DE images; and MALDI-TOF-MS was used to identify the differentially expressed proteins between the treated and untreated HL-60 cells. Western blot was performed to examine the expression level of Galectin-1 protein, one of the differential expression proteins. Results 2-DE patterns of the treated and untreated HL-60 cells with 5-aza-2dC were established, and a total of 35 differential protein spots were detected. Twenty non-redundant differential proteins were identified by MALDI-TOF-MS, of which 21 proteins were up-regulated and 6 proteins down-regulated after the 5-aza-2dC treatment. Western blot confirmed that the Galectin-1 protein was up-regulated after the 5-aza- 2dC treatment. Conclusion Encoding genes of 21 up-regulated proteins may be methylation-silencing genes in HL-60 cells, and be related to the pathogenesis of AML and 5-aza-2-dC treating AML.

关 键 词：蛋白质组学 二维凝胶电泳 5-杂氮-2’-脱氧胞苷 白血病 

分 类 号：R733.7[医药卫生—肿瘤]