幽门螺杆菌CagA对AGS细胞RNA转录相关磷酸化蛋白的影响  

The impact of CagA of Helicobacter pylori to phosphorylated RNA transcriptionassociated proteins in human gastric adenocarcinoma epithelial AGS cells

作　　者：肖迪[1] 赵飞[1] 宋衍燕[1] 孟凡亮[1] 何利华[1] 王海滨[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,北京102206

出　　处：《胃肠病学和肝病学杂志》2009年第4期332-336,共5页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家十一五科技支撑计划:中国幽门螺杆菌感染诊断和治疗的规范化研究(2007BAID4B02)

摘　　要：目的研究幽门螺杆菌(Helicobacter pylori,H.pylori)细胞毒素相关蛋白A(CagA)作用后人胃腺癌上皮细胞(AGS)RNA转录相关蛋白磷酸化的变化情况,进一步揭示CagA的致病机制,为寻找生物标记蛋白奠定基础。方法采用金属离子亲和吸附富集技术富集H.pylori、H.pyloriCagA缺失株(H.pylori△CagA)与AGS细胞相互作用4 h、以及培养相同时间的AGS细胞的磷酸化蛋白,利用二维凝胶电泳技术分离磷酸化蛋白,ImageMaster2D分析软件比较分析识别差异蛋白,4700型MALDI源蛋白分析器确认蛋白。结果CagA致使AGS细胞的7个RNA相关的磷酸化蛋白出现差异表达,其中1个磷酸化蛋白表达量上调、4个磷酸化蛋白表达量下调、2个新出现磷酸化的蛋白。hnRNP D0 127位的苏氨酸及137位的丝氨酸发生了磷酸化。结论CagA作用后,致使hnRNP D0蛋白磷酸化。AGS细胞与RNA转录相关蛋白磷酸化的变化,呈现出诱导细胞凋亡、增殖及有利于细胞免疫逃逸的趋势。Objective To study the change of RNA transcription-associated proteins in human gastric adenocareinoma epithelial AGS cells with CagA of Helicobacter pylori （H. priori） treatment, to reveal pathogenic mechanism of CagA and to lay the foundation of biomarker proteins. Methods Phosphorylated proteins in AGS cells were obtained by immobilized metal ion affinity chromatography after incubating H. priori 26695 strain, H. priori 26695 CagA deficient strains and AGS cells for four hours. Phosphorylated proteins were extracted using 2-dimensional polyacrylamide gel electrophoresis （2-DE） and analyzed by hnageMaster 2D analysis software. Proteins were confirmed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry （MALDI-TOF/TOF）. Results Seven RNA transcription-asso-ciated proteins altered expression in AGS cells by CagA. One was up-regulated. Four were down-regulated and two were phosphorylated, hnRNP D0 127 site threonine and 137 site serine were phosphorylated. Conclusion hnRNP D0 was phosphorylated after CagA treatment. The alternations of phosphorylated RNA transcription-associated proteins indicate a trend that induces cell apoptosis, proliferation and benefits cell immune escape.

关 键 词：磷酸化蛋白质 亲和层析 细胞毒素相关蛋白A 

分 类 号：R735.2[医药卫生—肿瘤] Q513.3[医药卫生—临床医学]