学术
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。[著者文摘]
文章出处:
《国际病理科学与临床杂志》-2007年27卷4期 -277-283页
栏目信息:
分 类 号:
文献标识码:
A
文章编号:
1673-2588（2007）04-0277-07
Differential proteomic analysis of 5-aza-2-dC treated and untreated nasopharyngeal carcinoma cells
ZHANG Wen-jing, YI Hong, LI Mao-yu, ZHANG Peng-Fei, LI Cui, TANG Cen-e, RUAN Lin, CHEN Zhu-chu, XIAO Zhi-qiang （ Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China）
Abstract:
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.[著者文摘]
Key words:
nasopharyngeal carcinoma （NPC） ; methylation silenced genes; two-dimension-al gel electrophoresis; mass spectrometry
基金资助:
教育部跨世纪优秀人才培养计划基金（教技函[2002]48）
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