Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

MKN74 gastric epithelial cells infected with Enterococcus faecalis for 24hrs or 5day

ABSTRACT: MKN74 cells were infected for 24 h or 5 days. For the 24 h infection, 80% confluent MKN74 cells were washed three times in PBS and incubated in antibiotic free medium. Overnight-grown colonies of E. faecalis were collected and added to the MKN74 cell culture at a multiplicity of infection (MOI) of 50 bacteria per cell. Cultures were maintained at 37 C under a 5% CO2 humidified atmosphere. 5 day infections were carried out by treating 65% confluent MKN74 cells with E. faecalis at a MOI of 10. Every 24 h, cells were washed three times with PBS and fresh medium and bacteria were added. Control cells were processed similarly in the absence of bacteria.

ORGANISM(S): Homo sapiens

SUBMITTER: Jesper Strickertsson

PROVIDER: E-MEXP-3496 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Enterococcus faecalis infection causes inflammation, intracellular oxphos-independent ROS production, and DNA damage in human gastric cancer cells.

Strickertsson Jesper A B JA Desler Claus C Martin-Bertelsen Tomas T Machado Ana Manuel Dantas AM Wadstrøm Torkel T Winther Ole O Rasmussen Lene Juel LJ Friis-Hansen Lennart L

PloS one 20130430 4

<h4>Background</h4>Achlorhydria caused by e.g. atrophic gastritis allows for bacterial overgrowth, which induces chronic inflammation and damage to the mucosal cells of infected individuals driving gastric malignancies and cancer. Enterococcus faecalis (E. faecalis) can colonize achlohydric stomachs and we therefore wanted to study the impact of E. faecalis infection on inflammatory response, reactive oxygen species (ROS) formation, mitochondrial respiration, and mitochondrial genetic stability ...[more]

PMID: 23646188

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Project description:Emerging antibiotic resistance among clinically relevant bacteria, paired with their ability to form biofilms on medical and technical devices, represents a serious problem in terms of effective and long-term decontamination in health care environments and gives rise to an urgent need for new antimicrobial materials. Here we present the first study of the impact of AGXXÂ®, a novel broad-spectrum antimicrobial surface coating consisting of micro galvanic elements formed by silver and ruthenium, on the transcriptome of the nosocomial pathogen Enterococcus faecalis. E. faecalis was subjected to metal stress by growing it for different periods of time in the presence of AGXXÂ® or silver-coated steel meshes. Subsequently, total RNA was isolated and next-generation RNA sequencing was performed to analyze variations in gene expression levels in the presence of the antimicrobial materials with focus on known stress genes. Exposure to AGXXÂ® had a large impact on the transcriptome of E. faecalis. After 24 minutes almost 1/5 of the E. faecalis genome displayed differential expression. At each time-point the cop operon was strongly up-regulated, providing indirect evidence for the presence of free Ag+-ions. Moreover, exposure to AGXXÂ® induced a broad general stress response in E. faecalis. Genes coding for the chaperones GroEL and GroES as well as the Clp proteases ClpE and ClpB were among the top up-regulated heat shock genes. Furthermore, differential expression of genes coding for thioredoxin, superoxide dismutase and glutathione synthetase indicates a high level of oxidative stress. We postulate a mechanism of action where the combination of Ag+-ions and reactive oxygen species generated by AGXXÂ® results in a synergistic antimicrobial effect, which is superior to that of conventional silver coatings. Gene expression analysis of Enterococcus faecalis 12030 either subjected to metal stress by exposure to an antimicrobial AGXXÂ®- or Ag-coated V2A steel mesh or exposed to an uncoated V2A steel mesh or left untreated performing RNA Sequencing with an Ion ProtonTM Sequencer and subsequent data analysis with a T-REx RNA-Sequencing expression analysis pipeline.

Dataset Information

MKN74 gastric epithelial cells infected with Enterococcus faecalis for 24hrs or 5day

Publications

Enterococcus faecalis infection causes inflammation, intracellular oxphos-independent ROS production, and DNA damage in human gastric cancer cells.

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