摘要目的 研究幽门螺杆菌(Hdicobacterpylori)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质谱答定确认蛋白.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伞面认识幽门螺杆菌的致病作用有重要意义.

abstractsObjective To research the differential 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 af-finity 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 ex-pressed 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. priori can cause a wide range change to human gastric adenocarcinoma epithelial cell protein pheshorylation. This change character has great significance to further comprehensive understanding of the pathogenesis of H. pylori.