MicroRNA expression in CagA-expressing cells
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ABSTRACT:
ORGANISM(S): Rattus norvegicus
PROVIDER: GSE27009 | GEO | 2011/04/21
SECONDARY ACCESSION(S): PRJNA137713
REPOSITORIES: GEO
Similar Datasets
Project description:Comprehensive analysis of microRNA expression were performed with 238 rat genes of microRNA between CagA-expressing and -non-expressing cells. CagA expression was controlled by tetracycline-off system in RGM1 cells. In the study presented here, RGM1 cells were used to acquire expression profiles of 238 microRNA genes.
2011-04-21 | E-GEOD-27009 | biostudies-arrayexpress
Project description:microRNA expression in CagA-expressing cells
| PRJNA137713 | ENA
Project description:The proteins from the Fanconi Anemia (FA) pathway of DNA repair maintain DNA replication fork integrity by preventing the unscheduled degradation of nascent DNA at regions of stalled replication forks. Here, we ask if the bacterial pathogen H. pylori exploits the fork stabilisation machinery to generate double stand breaks (DSBs) and genomic instability. Specifically, we study if the H. pylori virulence factor CagA generates host genomic DSBs through replication fork destabilisation and collapse. An inducible gastric cancer model was used to examine global CagA-dependent transcriptomic and proteomic alterations, using RNA sequencing and SILAC-based mass spectrometry, respectively. The transcriptional alterations were confirmed in gastric cancer cell lines infected with H. pylori. Functional analysis was performed using chromatin fractionation, pulsed-field gel electrophoresis (PFGE), and single molecule DNA replication/repair fiber assays. We found a core set of 31 DNA repair factors including the FA genes FANCI, FANCD2, BRCA1, and BRCA2 that were downregulated following CagA expression. H. pylori infection of gastric cancer cell lines showed downregulation of the aforementioned FA genes in a CagA-dependent manner. Consistent with FA pathway downregulation, chromatin purification studies revealed impaired levels of Rad51 but higher recruitment of the nuclease MRE11 on the chromatin of CagA-expressing cells, suggesting impaired fork protection. In line with the above data, fibre assays revealed higher fork degradation, lower fork speed, daughter strands gap accumulation, and impaired re-start of replication forks in the presence of CagA, indicating compromised genome stability. By downregulating the expression of key DNA repair genes such as FANCI, FANCD2, BRCA1, and BRCA2, H. pylori CagA compromises host replication fork stability and induces DNA DSBs through fork collapse. These data unveil an intriguing example of a bacterial virulence factor that induces genomic instability by interfering with the host replication fork stabilisation machinery.
2022-02-22 | PXD031168 | Pride
Project description:Helicobacter pylori infection promotes chronic inflammation and plethora of DNA damages including strand breaks, base alterations, point mutations, microsatellite instability and epigenetic alterations. The gastric epitheliaI cells, in order to prevent genomic instability, require an integrous DNA damage repair (DDR) machinery which, however, has been reported to be modulated by the infection. CagA is a major Hp virulence factor that deregulates host cell functions such as proliferation, apoptosis and chromosomal integrity. Its pathogenic activity is partly regulated by tyrosine phosphorylation on repeated EPIYA-motifs. Our aim was to identify putative effects of H. pylori infection and CagA protein on DDR machinery, investigating the transcriptome of AGS cells, infected with P12 H. pylori wild-type strain, as well as, its corresponding CagA knock-out and the CagA phosphorylation-defective mutant following tyrosine (EPIYA) substitution by phenylalanine (EPIFA) in the EPIYA-C motifs. Upon RNA-Sequencing on polyA-selected transcripts we performed Differential Expression Analysis, Pathway Enrichment Analysis and visualization on KEGG Pathway Maps per DDR mechanism. Key DDR components that were observed to be downregulated in a CagA-related manner were validated via Western Blot utilizing AGS and the non-cancerous GES1 cell lines. Transcriptome analysis revealed that a notable number of DDR genes were downregulated during H. pylori infection resulting to potential modulation of Base Excision Repair, Mismatch Repair and a more intricate deregulation of Nucleotide Excision Repair, Homologous Recombination and Non-Homologous End-Joining. CagA contributes to MUTYH, FEN1, APE1, POLD1 and LIG1 downregulation and those observations were verified on the protein expression level, with the exception of APE1 that was on contrary observed to be upregulated. Our study accentuates the role of CagA, as a significant contributor of the H. pylori infection-mediated DDR modulation, disrupting the balance between DNA damage introduction and repair thus favoring genomic instability and potentially contributing to gastric carcinogenesis.
2020-12-17 | GSE162056 | GEO
Project description:Although H.pylori infection is known to induce host genomic double strand breaks, (DSBs), the role of CagA, a central virulence factor with oncogenic properties in DSB generation remains unclear. Here, using transcriptomic and proteomic analyses, we show that CagA downregulates the expression of major DNA repair factors such as FANCD2, BRCA1 and BRCA2 of the Fanconi Anemia (FA) pathway of DNA repair and leads to compromised host replication fork speed, stability and toxic DSB induction. Our results highlight the mechanism of CagA-mediated genome instability at the level of replication forks and can be used to devise strategies that mitigate the genotoxic effects of H.pylori-infection.
2022-03-24 | GSE132356 | GEO
Project description:TC1: gastric epithelial (AGS) cells infected with wild type H. pylori (G27) and isogenic mutants in cagA and vacA for 0, 0.5, 3, 6, and 12 hours. Total RNA was used to make single stranded Cy5 labelled probe and compared to Cy3 labelled probe from uninfected AGS cells. Hybridizations of G27 (trial 4) and cagA- (trial 3) timecourses were done in parallel. A technical replicate of the G27 time course (trial 5) and hybridization of vacA- (trial 3) time course were done in parallel. The cagA 6 and 12 hour time points were technically replicated (trial 4) (the cagA 6 hour sample of trial 3 was lost). TC2: Biological replication and expansion of TC1, using more isogenic mutants and timepoints. AGS cells were mock infected, infected with G27, and isogenic mutants in cagN, cagA, cagE, and a deletion of the cag PAI for 0. 1, 3, 6, 12, and 24 hours. Probe synthesis and hybridization was done as in TC1. Note: there may have been a sample mix-up with PAI 12, swapping it with G27 or cagN 12.
2005-09-30 | E-SMDB-1661 | biostudies-arrayexpress
Project description:H. pylori virulence factors have been suggested to be important in determining the outcome of infection. The H. pylori adhesion protein BabA2 is thought to play an crucial role in bacterial colonization and in induction of a severe gastric inflammation, particularly in combination with expression of CagA and VacA. However, the influence of these virulence factors on the pathogenesis of H. pylori infection, is still poorly understood. To address this question, the inflammatory gene expression profiles from two groups of patients infected with triple-negative strains (lacking expression of CagA, BabA2 and VacAs1, but expressing VacAs2) and triple-positive strains (expressing CagA, VacAs1 and BabA2, but lacking expression of VacAs2) were investigated. The gene expression pattern in the antrum gastric mucosa from patients infected with different H. pylori strains was very similar, and no differentially expressed genes could be identified by pair-wise comparisons. Our data thus suggest a lack of correlation between the host inflammatory responses in the gastric mucosa and expression of the BabA2, CagA and VacAs1 genes. Two groups of patients who infected with double-positive strains (type I strains, expressing CagA and VacAs1 genes) and triple-negative strains (lacking the CagA, BabA2 and VacAs1, but expressing VacAs2) were investigated in this study.
2008-05-01 | E-GEOD-6143 | biostudies-arrayexpress
Project description:MicroRNA-seq of cell lines expressing pri-miR-9 paralog constructs
| PRJNA429035 | ENA
Project description:This study investigates the predictive and prognostic values of inflammatory markers and microRNA in stage IV colorectal cancer. The expression of inflammatory markers and microRNA in plasma will be correlated with tumor location, with dietary patterns and with survival during treatment.
| 2320890 | ecrin-mdr-crc
Project description:MicroRNA-seq of cell lines expressing pri-miR-9 paralog constructs II
| PRJNA723320 | ENA