Subcellular and functional proteomic analysis of the cellular responses induced by Helicobacter pylori
Subcellular and functional proteomic analysis of the cellular responses induced by Helicobacter pylori
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DOI：
10.1074/mcp.m500029-mcp200
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发表时间：
2006-04-01
影响因子：
7
通讯作者：
Chow, LP
中科院分区：
文献类型：
--
作者：
Chan, CH;Ko, CC;Chow, LP
幽门螺杆菌感染是几种消化系统疾病的重要致病因素，包括消化性溃疡、慢性胃炎和胃癌。此外，幽门螺杆菌可诱导胃上皮细胞表达疾病特异性蛋白。本研究的目的是鉴定幽门螺杆菌感染的胃上皮AGS细胞中差异表达的蛋白质。建立了一个体外模型，使用100个感染的复数，并通过功能分析来评估H.用蛋白质组学方法鉴定蛋白质图谱的变化，包括二维荧光差示凝胶电泳法和质谱仪。许多蛋白质的表达发生了变化，其中28个被鉴定为蛋白质合成和折叠相关蛋白、细胞骨架蛋白、代谢酶、转录和翻译相关蛋白、血管生成/转移相关蛋白、细胞通讯/信号转导相关蛋白或其他(氧调节蛋白和癌蛋白)。通过免疫印迹分析和免疫组织化学染色，进一步检测了层粘连蛋白-1链前体、Valosin含蛋白、热休克70 kDa蛋白、线粒体基质蛋白P1、FK506结合蛋白4、T-复合蛋白1、烯醇化酶α和14-3-3β等8种蛋白在癌组织和癌旁正常组织中的表达谱，以确定可能参与幽门螺杆菌诱导的胃病发病机制的分子靶点。根据我们的结果，含有Valosin的蛋白、线粒体基质蛋白P1、T-复合体蛋白1、烯醇化酶α和14-3-3β可能通过介导抗凋亡和增殖反应在Hp诱导的胃癌发生中起关键作用。
Helicobacter pylori infection is a crucial factor in the pathogenesis of several digestive disorders, including peptic ulcers, chronic gastritis, and gastric cancer. Moreover H. pylori induces disease-specific protein expression in gastric epithelial cells. The aim of the present study was to characterize proteins differentially expressed in H. pylori-infected gastric epithelial AGS cells. An in vitro model was established using a multiplicity of infection of 100 and evaluating the effectiveness of H. pylori infection by functional analyses. Changes in protein patterns were identified using a proteomic approach consisting of two-dimensional fluorescence difference gel electrophoresis and mass spectrometry. The expression of many proteins was found to be altered, and 28 of these were identified and classified as protein synthesis- and folding-related proteins, cytoskeleton proteins, metabolic enzymes, transcription- and translation-related proteins, angiogenesis/metastasis-related proteins, cell communication/signal transduction-related proteins, or others (oxygen-regulated protein and oncoprotein). The expression profiles of eight of these proteins, laminin gamma-1 chain precursor, valosin-containing protein, heat shock 70-kDa protein, mitochondrial matrix protein P1, FK506-binding protein 4, T-complex protein 1, enolase alpha, and 14-3-3 beta were further examined in cancerous and paired surrounding normal tissues by immunoblot assay and immunohistochemical staining to identify molecular targets that may be involved in the pathogenesis of H. pylori-induced gastric diseases. On the basis of our results, valosin-containing protein, mitochondrial matrix protein P1, T-complex protein 1, enolase alpha, and 14-3-3 beta may play a crucial role in H. pylori-induced gastric carcinogenesis by mediating antiapoptotic and proliferative responses.