5-aza-2-dC处理鼻咽癌细胞前后的差异蛋白质组学研究  

Differential proteomic analysis of 5-aza-2-dC treated and untreated nasopharyngeal carcinoma cells

作　　者：张文静[1] 易红[1] 李茂玉[1] 张鹏飞[1] 李萃[1] 汤参娥[1] 阮林[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》2007年第4期277-283,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(教技函[2002]48)~~

摘　　要：目的:比较5-杂氮-2′-脱氧胞苷(5-aza-2-dC)处理鼻咽癌细胞前后蛋白质组的差异,为筛选鼻咽癌的甲基化失活基因提供依据。方法:用去甲基化药物5-aza-2-dC处理鼻咽癌细胞系5-8F细胞,应用双向凝胶电泳(2-DE)技术分离5-aza-2-dC处理与未处理5-8F细胞的蛋白质,PDQuest图像分析软件识别药物处理与未处理5-8F细胞的差异表达蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定差异表达的蛋白质。Western印迹法检测差异蛋白质nm23-H1在药物处理与未处理5-8F细胞中的表达水平。结果:建立了5-aza-2-dC处理与未处理5-8F细胞蛋白质的2-DE图谱,识别了49个差异表达的蛋白质点,鉴定了33个差异表达的蛋白质,其中15个蛋白质在5-aza-2-dC处理5-8F后表达上调。Western印迹分析证实了nm23-H1在药物处理与未处理5-8F细胞中的差异表达水平。结论:15个5-aza-2-dC处理后表达上调蛋白质的编码基因可能是5-8F细胞的甲基化沉默基因。Objoetive To compare the proteome difference between nasopharyngeal carcinoma （NPC） cell line 5-8F cells treated and untreated with 5-aza-2-dC, and to provide experimental evidences for screening methylation silenced genes in NPC. Methods Two-dimensional gel electrophoresis （2- DE） was performed to separate proteins from treated and untreated 5-8F cells with 5-aza-2-dC, a demeth- ylation agent. PDQuest software was used to analyze 2-DE images, and MALDI-TOF-MS was used to identify the differentially expressed proteins between the 2 groups of 5-8F cells. The expression levels of differential protein nm23-H1 in the 2 groups of 5-8F cells lines were detected by Western blotting. Resuits 2-DE reference patterns of treated and untreated 5-8F cells with 5-aza-2-dC were established. Forty-nine differential protein spots were found between the 2 groups of 5-8F cells, a total of thirty-three non- redundant proteins were identified by MALDI-TOF-MS, and 15 proteins of them were up-regulated after 5-aza-2-dC treatment. The differential expression level of nm23-H1 in the 2 groups of 5-8F cells were confirmed by Western blotting. Conclusion Encoding genes of 15 up-regulated proteins after 5-aza-2-dC treatment may be methylation silenced genes in NPC cell line 5-8F cells.

关 键 词：鼻咽癌 甲基化沉默基因 双向凝胶电泳 质谱 

分 类 号：R739.63[医药卫生—肿瘤]