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dc.contributor.authorSigurdsson, Valgardur
dc.contributor.authorIngthorsson, Saevar
dc.contributor.authorHilmarsdottir, Bylgja
dc.contributor.authorGustafsdottir, Sigrun M
dc.contributor.authorFranzdottir, Sigridur Rut
dc.contributor.authorArason, Ari Jon
dc.contributor.authorSteingrimsson, Eirikur
dc.contributor.authorMagnusson, Magnus K
dc.contributor.authorGudjonsson, Thorarinn
dc.date.accessioned2014-05-20T09:33:39Z
dc.date.available2014-05-20T09:33:39Z
dc.date.issued2013-04
dc.date.submitted2014-05-20
dc.identifier.citationPLoS ONE 2013, 8(4):e60798en
dc.identifier.issn1932-6203
dc.identifier.pmid23573284
dc.identifier.doi10.1371/journal.pone.0060798
dc.identifier.urihttp://hdl.handle.net/2336/317190
dc.descriptionTo access publisher's full text version of this article. Please click on the hyperlink in Additional Links field.en
dc.description.abstractBranching morphogenesis is a mechanism used by many species for organogenesis and tissue maintenance. Receptor tyrosine kinases (RTKs), including epidermal growth factor receptor (EGFR) and the sprouty protein family are believed to be critical regulators of branching morphogenesis. The aim of this study was to analyze the expression of Sprouty-2 (SPRY2) in the mammary gland and study its role in branching morphogenesis. Human breast epithelial cells, breast tissue and mouse mammary glands were used for expression studies using immunoblotting, real rime PCR and immunohistochemistry. Knockdown of SPRY2 in the breast epithelial stem cell line D492 was done by lentiviral transduction of shRNA constructs targeting SPRY2. Three dimensional culture of D492 with or without endothelial cells was done in reconstituted basement membrane matrix. We show that in the human breast, SPRY2 is predominantly expressed in the luminal epithelial cells of both ducts and lobuli. In the mouse mammary gland, SPRY2 expression is low or absent in the virgin state, while in the pregnant mammary gland SPRY2 is expressed at branching epithelial buds with increased expression during lactation. This expression pattern is closely associated with the activation of the EGFR pathway. Using D492 which generates branching structures in three-dimensional (3D) culture, we show that SPRY2 expression is low during initiation of branching with subsequent increase throughout the branching process. Immunostaining locates expression of phosphorylated SPRY2 and EGFR at the tip of lobular-like, branching ends. SPRY2 knockdown (KD) resulted in increased migration, increased pERK and larger and more complex branching structures indicating a loss of negative feedback control during branching morphogenesis. In D492 co-cultures with endothelial cells, D492 SPRY2 KD generates spindle-like colonies that bear hallmarks of epithelial to mesenchymal transition. These data indicate that SPRY2 is an important regulator of branching morphogenesis and epithelial to mesenchymal transition in the mammary gland.
dc.description.sponsorshipLandspitali University Hospital Science Fund University of Iceland Research Fund Science and Technology Policy Council-Thematic program in postgenomic biomedicine Science and Technology Policy Council Research fund European Science Foundation (EuroCORES program, EuroSTELLS) Göngum saman a supporting group for breast cancer research in Icelanden
dc.language.isoenen
dc.publisherPublic Library of Scienceen
dc.relation.urlhttp://dx.doi.org/10.1371/journal.pone.0060798en
dc.relation.urlhttp://www.plosone.org/article/fetchObject.action?uri=info%3Adoi%2F10.1371%2Fjournal.pone.0060798&representation=PDFen
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3616012/pdf/pone.0060798.pdfen
dc.rightsopenAccessen
dc.subject.meshAnimalsen
dc.subject.meshCell Lineen
dc.subject.meshCell Movementen
dc.subject.meshCell Proliferationen
dc.subject.meshCoculture Techniquesen
dc.subject.meshEndothelial Cellsen
dc.subject.meshEpithelial-Mesenchymal Transitionen
dc.subject.meshFemaleen
dc.subject.meshGene Knockdown Techniquesen
dc.subject.meshHumansen
dc.subject.meshIntracellular Signaling Peptides and Proteinsen
dc.subject.meshLactationen
dc.subject.meshMammary Glands, Animalen
dc.subject.meshMammary Glands, Humanen
dc.subject.meshMiceen
dc.subject.meshMice, Inbred C57BLen
dc.subject.meshMorphogenesisen
dc.subject.meshPregnancyen
dc.subject.meshRNA, Small Interferingen
dc.subject.meshReceptor, Epidermal Growth Factoren
dc.subject.meshSignal Transductionen
dc.titleExpression and functional role of sprouty-2 in breast morphogenesis.en
dc.typeArticleen
dc.contributor.departmentUniv Iceland, Fac Med, BioMed Ctr, Stem Cell Res Unit, Reykjavik, Iceland Landspitali Univ Hosp, Dept Lab Hematol, Reykjavik, Iceland Univ Iceland, Fac Med, Dept Biochem & Mol Biol, Reykjavik, Iceland Univ Iceland, Fac Med, BioMed Ctr, Reykjavik, Iceland Univ Iceland, Fac Med, Dept Pharmacol, Reykjavik, Icelanden
dc.identifier.journalPloS oneen
dc.rights.accessOpen Accessen
refterms.dateFOA2018-09-12T13:19:34Z
html.description.abstractBranching morphogenesis is a mechanism used by many species for organogenesis and tissue maintenance. Receptor tyrosine kinases (RTKs), including epidermal growth factor receptor (EGFR) and the sprouty protein family are believed to be critical regulators of branching morphogenesis. The aim of this study was to analyze the expression of Sprouty-2 (SPRY2) in the mammary gland and study its role in branching morphogenesis. Human breast epithelial cells, breast tissue and mouse mammary glands were used for expression studies using immunoblotting, real rime PCR and immunohistochemistry. Knockdown of SPRY2 in the breast epithelial stem cell line D492 was done by lentiviral transduction of shRNA constructs targeting SPRY2. Three dimensional culture of D492 with or without endothelial cells was done in reconstituted basement membrane matrix. We show that in the human breast, SPRY2 is predominantly expressed in the luminal epithelial cells of both ducts and lobuli. In the mouse mammary gland, SPRY2 expression is low or absent in the virgin state, while in the pregnant mammary gland SPRY2 is expressed at branching epithelial buds with increased expression during lactation. This expression pattern is closely associated with the activation of the EGFR pathway. Using D492 which generates branching structures in three-dimensional (3D) culture, we show that SPRY2 expression is low during initiation of branching with subsequent increase throughout the branching process. Immunostaining locates expression of phosphorylated SPRY2 and EGFR at the tip of lobular-like, branching ends. SPRY2 knockdown (KD) resulted in increased migration, increased pERK and larger and more complex branching structures indicating a loss of negative feedback control during branching morphogenesis. In D492 co-cultures with endothelial cells, D492 SPRY2 KD generates spindle-like colonies that bear hallmarks of epithelial to mesenchymal transition. These data indicate that SPRY2 is an important regulator of branching morphogenesis and epithelial to mesenchymal transition in the mammary gland.


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