Dextran sodium sulfate-induced impairment of protein trafficking and alterations in membrane composition in intestinal Caco-2 cell line
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MDPI AG
Abstract
A key morphological feature of inflammatory bowel disease (IBD) is the loss of the barrier function of intestinal epithelial cells. The present study investigates endoplasmic reticulum (ER) stress in addition to alterations in protein and membrane trafficking in a dextran sulfate sodium (DSS)-induced IBD-like phenotype of intestinal Caco-2 cells in culture. DSS treatment significantly reduced the transepithelial electric resistance (TEER) and increased the epithelial permeability of Caco-2 cells, without affecting their viability. This was associated with an alteration in the expression levels of inflammatory factors in addition to an increase in the expression of the ER stress protein markers, namely immunoglobulin-binding protein (BiP), C/EBP homologous protein (CHOP), activation transcription factor 4 (ATF4), and X-box binding protein (XBP1). The DSS-induced ER-stress resulted in impaired intracellular trafficking and polarized sorting of sucrase-isomaltase (SI) and dipeptidyl peptidase-4 (DPPIV), which are normally sorted to the apical membrane via association with lipid rafts. The observed impaired sorting was caused by reduced cholesterol levels and subsequent distortion of the lipid rafts. The data presented confirm perturbation of ER homeostasis in DSS-treated Caco-2 cells, accompanied by impairment of membrane and protein trafficking resulting in altered membrane integrity, cellular polarity, and hence disrupted barrier function. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.
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Anti- and pro-inflammatory cytokines, Brush border membranes, Cholesterol, Dipeptidyl peptidase-4, Endoplasmic reticulum stress, Inflammatory bowel disease, Lipid rafts, Sucrase-isomaltase, Activating transcription factor 4, Alpha-glucosidases, Bacterial proteins, Caco-2 cells, Cell death, Cell membrane, Cell membrane permeability, Cell polarity, Cell survival, Cytokines, Dextran sulfate, Dipeptidyl peptidase 4, Epithelial cells, Humans, Inflammatory bowel diseases, Intestinal mucosa, Membrane microdomains, Protein transport, Sucrase-isomaltase complex, Transcription factor chop, X-box binding protein 1, Dipeptidyl peptidase iv, Glucose regulated protein 78, Growth arrest and dna damage inducible protein 153, Interleukin 10, Interleukin 1alpha, Interleukin 6, Lactate dehydrogenase, Sucrase isomaltase, X box binding protein 1, Alpha glucosidase, Atf4 protein, human, Bacterial protein, Cytokine, Ddit3 protein, human, Sucrase-isomaltase-maltase, Xbp1 protein, human, Article, Bioassay, Caco-2 cell line, Cell viability, Cholesterol level, Controlled study, Cytotoxicity, Electric resistance, Evans blue permeability assay, Fluorography, High performance liquid chromatography, Homeostasis, Human, Human cell, Immunoprecipitation, Lipid composition, Lipid raft, Phenotype, Protein analysis, Reverse transcription polymerase chain reaction, Rna isolation, Transepithelial electric resistance, Western blotting, Chemistry, Drug effect, Enzymology, Epithelium cell, Intestine mucosa, Membrane microdomain, Metabolism, Pathology, Physiology