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Glycosylation-related gene expression in HT29-MTX-E12 cells upon infection by Helicobacter pylori

World Journal of Gastroenterology 2017年第37期23卷 6817-6832页

作者： Michael T Cairns Ananya Gupta Julie A Naughton Marian Kane Marguerite Clyne lokesh joshi Glycoscience Group National Centre for Biomedical Engineering ScienceNational University of Ireland Galway School of Natural Sciences National University of Ireland Galway Conway Institute of Biomolecular and Biomedical Research School of Medicine and Medical Sciences University College Dublin

AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori(H. pylori).METHODS Polarised HT29-MTX-E12 cells were infected for 24 h with H. pylori strain 26695. After infection RNA was isolated from both infected and non-infected host cells. Sufficient infections were carried out to provide triplicate samples for microarray analysis and for q RTPCR analysis. RNA was isolated and hybridised to Affymetrix arrays. Analysis of microarray data identified genes significantly differentially expressed upon infection. Genes were grouped into gene ontology functional categories. Selected genes associated with host glycan structure(glycosyltransferases, hydrolases, lectins, mucins) were validated by real-time q RT-PCR *** Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. H. pylori do not survive incubation under the adopted culture conditions unless they associate with the adherent mucus layer of the host cell. Microarray analysis identified a total of 276 genes that were significantly differentially expressed(P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2. Six of these genes are involved in glycosylation-related processes. Real-time q RT-PCR demonstrated significant downregulation(1.8-fold, P < 0.05) of the mucin MUC20. REG4 was heavily expressed and significantly downregulated(3.1-fold, P < 0.05) upon infection. Gene ontology analysis was consistent with previous studies on H. pylori *** Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures.

关键词： Glycosylation Adherent mucus Gastric HT29-MTX-E12 H pylori strain 26695 Transcriptomics

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