不同转移潜能鼻咽癌细胞株的差异膜蛋白质组研究.pdfVIP

国际病理科学与临床杂志 Int J Pathol Clin Med 2012, 32(6) 479 不同转移潜能鼻咽癌细胞株的差异膜蛋白质组研究 陈瑜，李娇阳，李茂玉，肖志强 (中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室，长抄 410008) [摘 要] 目的:采用定量蛋白质组学技术筛选鼻咽癌转移相关的膜蛋白质。方法:在富集高转移鼻咽癌细 胞株 5 - 8F 和不转移鼻咽癌细胞株 6 一 10B 膜蛋白质的基础上，采用稳定同位素 180 标记结合串联 质谱技术比较两株细胞膜蛋白质组的差异，采用 Western 印迹对差异表达膜蛋白质 EphA2 的表达 水平进行验证，采用生物信息学方法对差异蛋白质进行 GO(gene oncology) 功能聚类和全基因组及 代谢途径数据库 (Kyoto encyclopedia of genes and genomes , KEGG) 信号通路分析。结果:鉴定了 31 个 5 - 8F 与 6 一 10B 细胞的差异膜蛋白，其中 25 个蛋白质在 5 - 8F 中表达上调、 6 个蛋白质表 达下调， Western 印迹技术验证了差异膜蛋白质 EphA2 的表达。生物信息分析显示，差异膜蛋白 的功能主要涉及细胞黠附、受体再循环和细胞连接等生物学过程，并参与了 14 个 KEGG 通路，其 中许多通路涉及细胞蒙古附、细胞连接以及细胞运动。结论: 31 个差异膜蛋白质可能在鼻咽癌转移 过程中发挥重要作用，为进一步研究鼻咽癌转移的分子机制提供了新线索。 [关键词] 鼻咽癌;转移;膜蛋白质组;定量蛋白质组学;180 标记 Differential membrane proteomic study of nasopharyngeal carcinoma cell lines with different metastatic potentials CHEN Yu, LI Jiaoyang, LI Mao归， XIAO Zhiqiang (均μ阳晒ryof臼ncerPro阳阳of胁呐01出lthofωm胁IgyaHo;等呼叫臼刷刷I Ui阳Jmity，α刷阱a41ω阴阳) Abstract Objective: To screen metastasis-related membrane proteins of nasopharyngeal carcinoma (NPC). Methods: A丘er enriching membrane proteins from high metastatic NPC cellline 5-8F and non-metastatic NPC cellline 6-lOB，由e post-digestion isotope 180 labeling method, followed by QTOF-MS/MS, was conducted to identify 也e differentially expressed membrane proteins in the 2 celllines. 1he 旺pression of differentia1 membrane protein EphA2 was measured by Western blot. Bioinformatics analysis was used to reveal the biological functions and Kyoto encyclopedia of genes and genomes (阻GG) pathways. Results: A total of 31 differential membrane proteins were identmed, of which 25 proteins were up-regulated in 5-8 F cells and 6 proteins were down-regu1ated. One of the differential proteins, EphA2, was further confirmed by Western blot. Bioinformatics analysis showed that the functions of the differential membrane proteins were mainly involved in cell adhesion, receptor recycling, and cell junction. The differential membrane proteins were also involved in 14 KEGG pat