Project description:Clopidogrel is associated with a series of gastrointestinal side effects, such as bleeding complications and recurrent gastric ulcer; however, their mechanisms are largely unclear. In this study, human gene expression microarray and gene ontology analysis were utilized to evaluate the effects of clopidogrel on gene expression in human gastric epithelial cells (GES-1) Keywords: Clopidogrel; Gastric injury; GES-1 cell line
Project description:Clopidogrel is associated with a series of gastrointestinal side effects, such as bleeding complications and recurrent gastric ulcer; however, their mechanisms are largely unclear. In this study, human gene expression microarray and gene ontology analysis were utilized to evaluate the effects of clopidogrel on gene expression in human gastric epithelial cells (GES-1) Keywords: Clopidogrel; Gastric injury; GES-1 cell line The Agilent Whole Human Genome Oligo Microarray was measured at 24 hours after GES-1 cells were exposure to clopidogrel ( 1.5mmol/l). The experimental set up the control group and the clopidogrel group. Three chips were performed on each group.
Project description:In this study, cultured H.pylori was used to infect the normal gastric epithelial cell line GES-1, to construct the H.pylori infection model. lncRNA microarray was applied for detecting the lncRNA in H.pylori infection model.
Project description:Research on lncRNAs expression profiles in human interactions with Helicobacter pylori (H.pylori) is rare reported. We used HTA2.0 to investigate the expression changes of lncRNAs and mRNAs in human gastric epithelial cells (GES-1 cell line) infected with or without H.pylori infection. Our study provided a preliminary exploration of lncRNAs expression profiles in H.pylori-infected cell models by microarray. A subset of aberrantly expressed lncRNAs was verified by qRT-PCR. These dysregulated lncRNAs might contribute to the pathological processes during H.pylori infection.
Project description:Research on lncRNAs expression profiles in human interactions with Helicobacter pylori (H.pylori) is rare reported. We used HTA2.0 to investigate the expression changes of lncRNAs and mRNAs in human gastric epithelial cells (GES-1 cell line) infected with or without H.pylori infection. Our study provided a preliminary exploration of lncRNAs expression profiles in H.pylori-infected cell models by microarray. A subset of aberrantly expressed lncRNAs was verified by qRT-PCR. These dysregulated lncRNAs might contribute to the pathological processes during H.pylori infection. We infected GES-1 cells with H.pylori as experimental groups,and cells without H.pylori infection were regarded as control groups. After 24hr-infection, cells of each group were selected for total RNA extraction and hybridization on Affymetrix HTA2.0 arrays. The aberrant expression profiles of lncRNAs and mRNAs were explored by microarray analysis between experimental and control groups.
Project description:Mitochondria exhibit diverse effects on cellular responses. Mitochondrial transplantation from gastric epithelial cells GES-1 to gastric cancer cells AGS reduces cancer malignancy was previously reported. We employed proteomic and metabolomic analyses to elucidate underlying mechanisms. iTRAQ/TMT-based proteomics identified 257 upregulated and 34 downregulated proteins, with Ingenuity pathway analysis revealing regulation of 14 signaling pathways. The upregulation of p53, Bax, p-AktS473, p-mTORS2448 and the down-regulation of Sirt 3, p-NRF2S40, and HO-1 were further verified by western blotting. Metabolomic profiling detected 3 upregulated and 8 down-regulated metabolites implicated in glycolysis, TCA cycle, pentose phosphate pathway (PPP), and ATP production. Further investigation showed that decreased extracellular lactate correlating with enhanced MCT1 expression (lactate importer), reduced MCT4 expression (lactate exporter), and increased LDHB expression (lactate to-pyruvate conversion). With the finding that transplanted with GES-1 mitochondria and induced accumulation of pyruvate, the AGS was solely treated with pyruvate and the result showed a cell migration of AGS was retarded. Additionally, isocitrate accumulation preceded decreased α-ketoglutarate, malate, ATP, and NADH levels. In conclusion, integrated proteomic and metabolomic analyses revealed that transplanted GES-1 mitochondria attenuate AGS gastric cancer malignancy through stagnation of glycolysis and the TCA cycle progression, highlighting potential targets for mitochondrial-based therapies.
Project description:To analyze the gene expression of human gastric epithelial cells (GES-1) regulated by paeonifl or in based on transcript tome sequencing; to identify and analyze key genes; to study the mechanism of paeoniflorin on Leuk-1cells; and to explore its medicinal value. Method: Different doses of Ulcer-healing Decoction were used to treat GES-1, and its activity was assessed by MTT assay. Transcriptome sequencing analysis was then performed to identify differentially expressed genes (DEGs). The function and pathways of the DEGs were annotated using GO and KEGG enrichment analysis. Result: The survival rate of GES-1 cells was significantly increased under different concentrations of ulcer-healing decoction intervention (P < 0.05 or P < 0.01). After 24 hours of treatment with 80 µ g/mL ulcer-healing decoction, a total of 312 crucial DEGs were observed in GES-1 cells, of which 233 DEGs were upregulated and 79 DEGs were downregulated. The top five differentially expressed genes include CYP1A1, IL24, IKBKGP1, GDF15, SCUBE1 and OASL, ADAMTS5, SULF1, ID2, and DDX60. The functions of these genes are mainly related to promoting wound,cell proliferation and migration,immunity and inflammation.GO enrichment analysis showed that the DEGs were significantly enriched in the extracellular region and involved in biological processes such as cytokine production and response, stimulus response, and exerting molecular functions such as proliferation factor receptor activity. KEGG pathway enrichment showed that the DEGs were significantly enriched in various signaling pathways. Immunohistochemistry and other methods were used to detect differentially expressed proteins in GES-1 cells induced by ulcer-healing decoction, and the results were consistent with those from RNA-seq, thereby validating the accuracy of sequencing technologies. Conclusion: The role of ulcer-healing decoction in GES-1 is primarily reflected in regulating inflammatory factors and growth factors, promoting cell proliferation, and supporting the regeneration and repair of gastric mucosal epithelial cells.
Project description:With the colonization of Helicobacter Pylori (H.pylori) in the human stomach, a large amount of Helicobacter Pylori (H.pylori)colonized into the stomach disintegrates and releases a large number plenty of toxic proteins and other bacterial structure components. TollToll-like receptors (TLRs) in gastric epithelial cells which sensitive to the components of bacteria lysate around the microenvironment attach them and activite innate immune and pro-inflammatory response once contact with antigen. The consequences after long-term stimulation of H.pylori lysate in gastric epithelial cells has not been reported . To investigate the effects of H.pylori lysate on TLRs expression and inflammatory cytokines level during long-term stimulation of gastric epithelial cells and clarify the potential mechanismHence, we detected TLRs mRNA level and inflammatory cytokines expression in GES-1 cells co-culture with lysate from 1 to 30 generations. The sensitivity to H.pylori or H.pylori lysate in GES-1 cells were also monitored. We also explored whether LPS was the main factors contributing to this change. Mongolian gerbils were used to replicate H.pylori infection animal model. After transcriptome sequencing,A TLR2/6 heterodimers agonist Pam2CSK4 and a highly selective inhibitor of PI3K LY294002 were used to clarify the TLR6 signaling pathways. In a conclusion, we discovered that the sensitivity of TLRs in human gastric epithelial cells decreased while after long-time co-culture with H.pylori lysate. Human gastric epithelial cells shows tolerance to H.pylori lysate in TLRs and inflammatory cytokines level. decreased the high level of TLRs and inflammatory cytokines after stimulating GES-1 cells with H.pylori lysate for a short-term. And Tthis process may works through the TLR6/PI3K/NF-κKB axis.