title: p38δ mitogen-activated protein kinase regulates the expression of tight junction protein ZO-1 in differentiating human epidermal keratinocytes.
abstract: Increasing evidence has recognized tight junctions (TJs) as the lower epidermal inside-out diffusion barrier located in granular cell layers of the epidermis. However, little is known about the regulation of TJ components in epidermis. p38 pathway is one of the mitogen-activated protein kinase pathways, which controls cell growth, differentiation, and apoptosis. We have investigated the role of p38 signaling pathway in the regulation of selected desmosomal, adherens and TJ components in human primary keratinocytes during Ca(2+)-induced differentiation, as well as in cultured squamous cell carcinoma cell lines. p38 signaling pathway was inhibited in cultured keratinocytes and cutaneous squamous cell carcinoma cells using recombinant adenoviruses, small inhibitory RNAs (siRNA) and chemical inhibitors. Expression of intercellular junction proteins was investigated using Western analysis and indirect immunofluorescence (IIF). The results showed that inhibition of p38δ function by siRNA or adenovirally delivered dominant negative mutant led to markedly decreased levels of Zonula occludens-1 (ZO-1) protein in keratinocytes, while the expression of other junctional proteins studied was not altered. Immunolocalization of ZO-1 revealed that intercellular junction areas were depleted from ZO-1. Inhibition of ZO-1 by siRNA silencing did not however result in an altered expression or subcellular localization of other TJ components studied. The expression of ZO-1 in carcinoma cells was also regulated by p38. The results indicate that ZO-1 is regulated by p38δ while the other junction proteins studied are not. Since ZO-1 is an integral component of functional TJs, various pathological processes affecting signaling via p38δ may also interfere with epithelial maturation and the formation and function of TJs.
authors.affiliation: Department of Dermatology, University of Turku and Turku University Hospital, P.O.B 52, 20521, Turku, Finland.
authors.forename: Elina
authors.initials: E
authors.lastname: Siljamäki
authors.forename: Laura
authors.initials: L
authors.lastname: Raiko
authors.forename: Mervi
authors.initials: M
authors.lastname: Toriseva
authors.forename: Liisa
authors.initials: L
authors.lastname: Nissinen
authors.forename: Tuomas
authors.initials: T
authors.lastname: Näreoja
authors.forename: Juha
authors.initials: J
authors.lastname: Peltonen
authors.forename: Veli-Matti
authors.initials: VM
authors.lastname: Kähäri
authors.forename: Sirkku
authors.initials: S
authors.lastname: Peltonen
mesh_terms: D000328:Adult; D000368:Aged; D002118:Calcium / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000473:pathology; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D015603:Keratinocytes / Q000187:drug effects / Q000502:physiology*; D008875:Middle Aged; D048292:Mitogen-Activated Protein Kinase 13 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction; D019108:Tight Junctions / Q000378:metabolism*; D055815:Young Adult; D062826:Zonula Occludens-1 Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D007093:Imidazoles; D011725:Pyridines; D034741:RNA, Small Interfering; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; D048292:Mitogen-Activated Protein Kinase 13; C093642:SB 203580; D002118:Calcium