title: Histone deacetylase inhibitors up-regulate the expression of tight junction proteins.
abstract: Histone deacetylase (HDAC) inhibitors promote cell maturation, differentiation, and apoptosis through changes in gene expression. Differentiated epithelial cells are characterized by apical tight junctions (TJ), which play a role in cell-cell adhesion, polarity, and the permeability barrier function of epithelia. The relationship between cellular differentiation and expression of TJ-associated proteins is not known. Here, we investigated whether HDAC inhibitors affect the expression of TJ proteins in cultured cells by immunoblotting, immunofluorescence, and quantitative real-time, reverse transcription-PCR. We find that the HDAC inhibitor sodium butyrate significantly up-regulates the protein levels of cingulin, ZO-1, and ZO-2 in Rat-1 fibroblasts, cingulin in COS-7 cells, and cingulin and occludin in HeLa cells. Levels of mRNA for cingulin, ZO-1, and ZO-2 are also increased in sodium butyrate-treated Rat-1 fibroblasts. Up-regulation of cingulin is reversible and dose dependent and requires de novo protein synthesis and protein kinase activity, because it is inhibited by cycloheximide and by the protein kinase inhibitor H-7. Up-regulation of TJ proteins by sodium butyrate is linked to the ability of sodium butyrate to inhibit HDAC activity, because suberoylanilide hydroxamic acid, a HDAC inhibitor of a different structural class, also up-regulates cingulin, ZO-1, and ZO-2 expression in Rat-1 fibroblasts. These results indicate that cellular differentiation correlates with kinase-dependent up-regulation of the expression of specific TJ proteins.
authors.affiliation: Department of Molecular Biology, University of Geneva, 30 Quai Ernest Ansermet, 1211 Geneva 4, Switzerland.
authors.forename: Mauro
authors.initials: M
authors.lastname: Bordin
authors.forename: Fabio
authors.initials: F
authors.lastname: D'Atri
authors.forename: Laurent
authors.initials: L
authors.lastname: Guillemot
authors.forename: Sandra
authors.initials: S
authors.lastname: Citi
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology; D016475:3T3 Cells; D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D019556:COS Cells; D002448:Cell Adhesion; D002454:Cell Differentiation; D003513:Cycloheximide / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005347:Fibroblasts / Q000378:metabolism; D006367:HeLa Cells; D056572:Histone Deacetylase Inhibitors; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D015151:Immunoblotting; D058610:Isobutyrates; D008565:Membrane Proteins / Q000096:biosynthesis; D051379:Mice; D008840:Microfilament Proteins; D008856:Microscopy, Fluorescence; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019108:Tight Junctions; D015854:Up-Regulation; D000077337:Vorinostat
chemical_list: D002087:Butyrates; C061407:CGN protein, human; D004791:Enzyme Inhibitors; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D058610:Isobutyrates; D008565:Membrane Proteins; D008840:Microfilament Proteins; D047428:Protein Kinase Inhibitors; D000077337:Vorinostat; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C020380:isobutyric acid; D003513:Cycloheximide