Dataset Information

The quantitative proteomic analysis of extracellular vesicles derived from gastric cell line infected with Helicobacter pylori Tx30a, TN2wt strains, and non-infection

ABSTRACT: Human gastric adenocarcinoma AGS cells were infected with H. pylori Tx30a or TN2wt at a multiplicity of infection = 100 and incubated at 5% CO2,37oC for 48 h. The cells/debris were removed from the cell culture supernatant by centrifugation at 2,000 x g for 30 min at 4oC and filtered through a 0.45 m filter. The EVs were collected by ultracentrifugation at 110,000 x g for 70 min using Himac CP80NX Preparative Ultracentrifuge (HITACHI, Tokyo, Japan). After removing the supernatant, the EVs were washed with PBS and then collected by ultracentrifugation at 100,000 x g for 70 min at 4oC. Finally, the EVs were resuspended in PBS, and stored at -80oC. The AGS-derived EVs were lysed with 2% sodium dodecyl sulfate/ 7M urea, and equal amounts of proteins from each sample were precipitated with acetone. The proteins were then denatured and reduced with 50% trifluoroethanol and 4 mM dithiothreitol followed by alkylation of the free cysteine residues and trypsinization. The desalted peptides were then separated using liquid chromatography. The mass spectrometer was operated in information-dependent acquisition mode, and acquired spectra were searched against the AGS cell protein database. Differential protein expression was analyzed to determine the relative abundances of proteins in EVs derived from AGS cells infected with H. pylori Tx30a or TN2wt, compared to non-infected cells. H. pylori proteins in AGS-derived EV, the spectra were searched against the H. pylori ATCC 51932 (Tx30a) and TN2wt protein database.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Krisana Asano

PROVIDER: PXD056529 | JPOST Repository | Mon Mar 17 00:00:00 GMT 2025

REPOSITORIES: jPOST

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Dataset's files

Source:

			Action	DRS
	230919_Phawinee_AGS-TN2_1_SWATH.wiff	Wiff
	230919_Phawinee_AGS-TN2_1_SWATH.wiff.scan	Wiff
	230919_Phawinee_AGS-TN2_2_SWATH.wiff	Wiff
	230919_Phawinee_AGS-TN2_2_SWATH.wiff.scan	Wiff
	230919_Phawinee_AGS-TN2_3_SWATH.wiff	Wiff

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Publications

Proteomic Profiling of Extracellular Vesicles Reveals Potential Biomarkers for Helicobacter pylori Infection and Gastric Cancer.

Subsomwong Phawinee P Asano Krisana K Akada Junko J Matsumoto Takashi T Nakane Akio A Yamaoka Yoshio Y

Helicobacter 20250301 2

<h4>Background</h4>Helicobacter pylori (H. pylori) has been identified as a type I carcinogen and contributes to a high rate of gastric cancer (GC), especially in Eastern Asia. Extracellular vesicles (EVs) have the potential to be used to detect various cancer types and diseases. However, the protein markers in EVs for the prognosis of H. pylori infection and GC are unknown. We aim to identify the proteins within EVs derived from a gastric epithelial cell line (AGS) infected with H. pylori by us ...[more]

PMID: 40033163

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Project description:Studying the proteomes of tissue-derived extracellular vesicles (EVs) can lead to the identifica-tion of biomarkers of disease and can provide a better understanding of cell-to-cell communica-tion in both healthy and diseased tissue. The aim of this study was to apply our previously es-tablished tissue-derived EV isolation protocol to mouse lungs in order to determine the changes in the proteomes of lung tissue-derived EVs during allergen-induced eosinophilic airway in-flammation. A mouse model for allergic airway inflammation was used by sensitizing the mice intraperitoneal with ovalbumin (OVA), and one week after the final sensitization, the mice were challenged intranasal with OVA or PBS. The animals were sacrificed 24 h after the final chal-lenge, and their lungs were removed and sliced into smaller pieces that were incubated in cul-ture media with DNase I and Collagenase D for 30 min at 37 °C. Vesicles were isolated from the medium by ultracentrifugation and bottom-loaded iodixanol density cushions, and the proteo-mes were determined using quantitative mass spectrometry. More EVs were present in the lungs of the OVA-challenged mice compared to the PBS-challenged control mice. In total, 4510 proteins were quantified in all samples. Among them, over 1000 proteins were significantly altered (fold change >2), with 614 proteins being increased and 425 proteins being decreased in the EVs from OVA-challenged mice compared to EVs from PBS-challenged animals. The associated cellular components and biological processes were analyzed for the altered EV proteins, and the proteins enriched during allergen-induced airway inflammation were mainly associated with gene on-tology (GO) terms related to immune responses. In conclusion, EVs can be isolated from mouse lung tissue, and the EVs’ proteomes undergo changes in response to allergen-induced airway in-flammation. This suggests that the composition of lung-derived EVs is altered in diseases asso-ciated with inflammation of the lung, which may have implications in type-2 driven eosino-philic asthma pathogenesis.