Korean J Gastroenterol.  2014 Jul;64(1):10-17. 10.4166/kjg.2014.64.1.10.

Proteomics Analysis for Helicobacter pylori-infected Gastric Mucosa

• Kang HS
• Hong SN
• Park HR
• Kwon MJ
• Lee JH
• Kim JJ

• KMID: 1791889
• DOI: http://doi.org/10.4166/kjg.2014.64.1.10

Abstract

BACKGROUND/AIMS
Helicobacter pylori infection is linked to the development of gastric cancer. H. pylori-associated gastric inflammation is considered to be the first important step in the histogenesis of such neoplasia. However, studies that compare proteome of gastric mucosa infected with or without H. pylori are lacking.
METHODS
We employed proteomics analysis on the endoscopic biopsy specimens of gastric mucosa obtained from two groups (30 cases): healthy subjects without H. pylori infection (15 cases), and gastritis patients with H. pylori infection (15 cases). The pooled proteins obtained from gastric mucosa infected with or without H. pylori were separated by two-dimensional gel electrophoresis and analyzed by a computer-aided program. The altered protein expressions were then identified by mass spectrometry and validated by Western blotting and immunohistochemistry.
RESULTS
On mass spectrometry using MALDI TOF(TM) Analyzer, the up-regulation of Keratin 1, ezrin, adenosine triphosphate (ATP) synthase subunit alpha mitochondrial isoform c, Keratin type I cytoskeletal 19, and Keratin type I cytoskeletal 9 were identified; in contrast, 71 kd heat shock cognate protein, ATP synthase subunit alpha mitochondrial precursor, and annexin IV were down-regulated. Among them, membrane cytoskeleton linker ezrin was validated using Western blot and immunohistochemistry.
CONCLUSIONS
Expression of ezrin was significantly different between the gastric mucosa with and without H. pylori infection. Therefore, ezrin could be considered a promising potential molecular marker for detecting H. pylori infection in gastric mucosa.

Keyword

Helicobacter pylori; Proteomics; Ezrin; Stomach neoplasms; Gastritis

MeSH Terms

Blotting, Western
Cytoskeletal Proteins/metabolism
Down-Regulation
Electrophoresis, Gel, Two-Dimensional
Female
Gastric Mucosa/*metabolism/microbiology
Gastritis/complications/metabolism/pathology
Gastroscopy
Helicobacter Infections/complications/metabolism/*pathology
*Helicobacter pylori
Humans
Immunohistochemistry
Male
Proteome/*analysis
*Proteomics
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Up-Regulation
Cytoskeletal Proteins
Proteome

Figure

• 

  Fig. 1. Two-dimensional gel electrophoresis profiles of Helicobacter pylori(−) stomach and H. pylori(＋) nodular gastritis. A total of 100  g of whole-cell protein from each strain was applied on 170-mm immobilized pH gradient strips with a range of pH 3–10, followed by 12% sodium dodecyl sulfate (SDS-PAGE), and visualized by silver staining (up-regulated 5 protein spots [round] and down-regulated 5 protein spots [square] in H. pylori infected gastric mucosa are indicated at figure). IEF, isoelectric focusing.

• 

  Fig. 2. Up- (A) and down (B)-regulated protein spots in Helicobacter pylori infected gastric mucosa of 2-dimensional gel electrophoresis map. Circles indicate the differentially expressed proteins whose expression level was at least more than two times higher than the other group. The expression level was determined by the relative spot volume of the proteins compared to the total amount of the protein in the gel, and is expressed as the percentage volume (right panel).

• 

  Fig. 3. Western blot for ezrin in gastric mucosa infected with and without Helicobacter pylori.

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  Fig. 4. Immunohistochemistry for ezrin in gastric mucosal infected with and without Helicobacter pylori (×100).

Reference

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