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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。
OBJECTIVE: To compare the differences of proteome between 5-aza-2’-deoxycytidine (5-aza-2-dC) and nasopharyngeal carcinoma cells before and after treatment, and to provide basis for screening methylated inactivation genes of nasopharyngeal carcinoma. Methods: Nasopharyngeal carcinoma cell line 5-8F was treated with demethylation drug 5-aza-2-dC. The 5-aza-2-dC and untreated 5-aza- -8F cells, and PDQuest image analysis software to identify differentially expressed protein spots in drug-treated and untreated 5-8F cells and to identify differentially expressed proteins by matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF-MS). Western blotting was used to detect the expression level of nm23-H1 in drug-treated and untreated 5-8F cells. RESULTS: A 2-DE map of 5-aza-2-dC-treated and untreated 5-8F cell proteins was established, 49 differentially expressed protein spots were identified and 33 differentially expressed proteins identified, 15 of which The expression of 5-aza-2-dC was up-regulated after 5-8F treatment. Western blot analysis confirmed the differential expression level of nm23-H1 in drug-treated and untreated 5-8F cells. CONCLUSION: The gene encoding 15-aza-2-dC upregulated protein may be the methylation-silencing gene of 5-8F cells.