幽门螺杆菌感染后人胃腺癌上皮细胞微量磷酸化蛋白质差异分析  

Phosphoproteomic analysis of differential expression of AGS cellular proteins in response to Helicobacterpylori infection

作　　者：肖迪[1] 宋衍燕[2] 赵飞[1] 何利华[1] 孟凡亮[1] 张慧芳[1] 张建中[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所诊断室,北京102206 [2]北京市朝阳区疾病预防控制中心,100021

出　　处：《中华微生物学和免疫学杂志》2009年第5期449-453,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家十一五科技支撑计划（2007BAID4B02）

摘　　要：目的研究幽门螺杆菌（Helicobacter pylori）作用后人胃腺癌上皮细胞（AGS）中微量磷酸化蛋白的变化情况。方法采用金属离子亲和吸附富集技术富集幽门螺杆菌与AGS细胞相互作用以及AGS细胞的磷酸化蛋白，除盐纯化后利用二维凝胶电泳技术分离磷酸化蛋白，MALDI—TOF／TOF质谱鉴定确认蛋白。结果幽门螺杆菌作用后AGS细胞有21种蛋白出现差异表达，其中15种蛋白表达量下调，4种蛋白出现新表达，1种蛋白表达量上调，1种蛋白不表达。差异表达蛋白涉及细胞的钙离子调节、转录、翻译、蛋白折叠与运输、核糖体的装配、中心体复制、染色体稳定、细胞结构形态、细胞增殖与凋亡等。结论幽门螺杆菌能引起广泛的胃腺癌黏膜上皮细胞蛋白磷酸化特征的变化，这种变化谱特征对进一步全面认识幽门螺杆菌的致病作用有重要意义。Objective To research the differenti-al expression of trace phosphorylated proteins in human gastric adenocarcinoma epithelial （AGS） cells infected by Helicobacter pylori. Methods H. pylori 26695 strain infected AGS cells 4 h and AGS cells was cultivated for 4 h as a comparison. The proteins of AGS and comparison AGS cells were extracted. Their phosphorylated proteins were enriched by metal ion affinity adsorption enrichment techniques. After desalinated and purified the phosphorylated proteins samples were separated by 2-dimensional polyacrylamide gel electrophoresis （2-DE） technique. Computer assisted image analysis was used to analyze the differential proteomic expression. The significantly differentially expressed proteins were unambiguously assigned identities by matrix-assisted laser desorption/ionization-time of flight mass spectrometry （MALDI-TOF/TOF）. Results Fifteen kinds of proteins were down-regulated, 4 kinds of new proteins were observed, 1 kind of proteins were up-regulated, 1 kind of proteins unexpression. The 21 proteins that were significantly differentially expressed , including cellular calcium ion homeostasis, transcription, interpretation, protein folding and transport, ribosomal assembly, centrosome replication, chromosome stability, cellular structure, cellular proliferation and apoptosis. Conclusion H. pylori can cause a wide range change to human gastric adenocarcinoma epithelial cell protein phoshorylation. This change character has great significance to further comprehensive understanding of the pathogenesis of H. pylori.

关 键 词：亲和吸附 磷酸化蛋白 二维凝胶电泳 致病机制 

分 类 号：R735.2[医药卫生—肿瘤]