Subcellular and Functional Proteomic Analysis of the Cellular Responses Induced by Helicobacter pylori

Abstract

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 ␥-1 chain precursor, valosin-containing protein, heat shock 70-kDa protein, mitochondrial matrix protein P1, FK506-binding protein 4, Tcomplex protein 1, enolase ␣, and 14-3-3 ␤ 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 ␣, and 14-3-3

Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value

“…Cultures of the biopsy tissue samples used to isolate Helicobacter pylori were performed on Tryptic Soy Blood Agar with 5% defribinated sheep blood under microaerophilic conditions (10% CO2, 5% oxygen, 85% in air) at 37ºC for 5 to 7 days (Andersen and Espersen, 1992;Chan et al, 2006). Then, the samples were subcultured to isolate single colonies.…”

“…The resulting samples were cleaned up using ZipTipC18 pipette tips (Milipore). Finally, samples from the Zip-Tip were eluted into clean micro-centrifuge tubes and concentrated using a vacuum concentrator until they were completely dry (Eppendorf, USA), at this stage, the samples were ready for MALDI-TOF/ TOF mass analysis (Baik et al, 2004;Chan et al, 2006).…”

Biomarkers and diagnostic tools for detection of Helicobacter pylori

“…More than 90 % of population in developing and Eastern countries and half of the world population have been affected with H. pylori infection (Chan et al 2006a;Malaty 2007a). The prevalence rate of infection during the first decade and by the sixth decade of life in industrialized countries (West Europe and North America) is approximately 10 and 60 %, respectively.…”

“…This protein is related to the increased virulence and risk of gastric carcinoma. CagA is encoded by a gene placed in the Cag pathogenicity island (Cag PAI), which is found in more than 90 % of H. pylori isolated in Eastern countries and almost 60 % in Western countries (Chan et al 2006b). The CagPAI is 40 kb in size and a complex of Cag genes including CagI, CagE, CagL, CagH, and CagM, which are a major virulence factor of H. pylori (Chan et al 2006a;Cui and Shao 2007).…”

“…CagA is encoded by a gene placed in the Cag pathogenicity island (Cag PAI), which is found in more than 90 % of H. pylori isolated in Eastern countries and almost 60 % in Western countries (Chan et al 2006b). The CagPAI is 40 kb in size and a complex of Cag genes including CagI, CagE, CagL, CagH, and CagM, which are a major virulence factor of H. pylori (Chan et al 2006a;Cui and Shao 2007). H. pylori makes this protein via a type IV secretion system and physically injects it into the mucosal cells using its pilus (Fleming et al 2009;Peek et al 2010).…”