PMID- 23573284
OWN - NLM
STAT- MEDLINE
DCOM- 20131022
LR  - 20211021
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 8
IP  - 4
DP  - 2013
TI  - Expression and functional role of sprouty-2 in breast morphogenesis.
PG  - e60798
LID - 10.1371/journal.pone.0060798 [doi]
LID - e60798
AB  - Branching 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.
FAU - Sigurdsson, Valgardur
AU  - Sigurdsson V
AD  - Stem Cell Research Unit, BioMedical Center, Faculty of Medicine, University of 
      Iceland, Reykjavik, Iceland.
FAU - Ingthorsson, Saevar
AU  - Ingthorsson S
FAU - Hilmarsdottir, Bylgja
AU  - Hilmarsdottir B
FAU - Gustafsdottir, Sigrun M
AU  - Gustafsdottir SM
FAU - Franzdottir, Sigridur Rut
AU  - Franzdottir SR
FAU - Arason, Ari Jon
AU  - Arason AJ
FAU - Steingrimsson, Eirikur
AU  - Steingrimsson E
FAU - Magnusson, Magnus K
AU  - Magnusson MK
FAU - Gudjonsson, Thorarinn
AU  - Gudjonsson T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130403
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Membrane Proteins)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (SPRY2 protein, human)
RN  - EC 2.7.10.1 (EGFR protein, human)
RN  - EC 2.7.10.1 (ErbB Receptors)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cell Movement
MH  - Cell Proliferation
MH  - Coculture Techniques
MH  - Endothelial Cells/physiology
MH  - Epithelial-Mesenchymal Transition
MH  - ErbB Receptors/metabolism
MH  - Female
MH  - Gene Knockdown Techniques
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins/genetics/*metabolism
MH  - Lactation
MH  - Mammary Glands, Animal/physiology
MH  - Mammary Glands, Human/*growth & development/metabolism
MH  - Membrane Proteins
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - *Morphogenesis
MH  - Pregnancy
MH  - RNA, Small Interfering/genetics
MH  - Signal Transduction
PMC - PMC3616012
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2013/04/11 06:00
MHDA- 2013/10/23 06:00
PMCR- 2013/04/03
CRDT- 2013/04/11 06:00
PHST- 2012/07/22 00:00 [received]
PHST- 2013/03/03 00:00 [accepted]
PHST- 2013/04/11 06:00 [entrez]
PHST- 2013/04/11 06:00 [pubmed]
PHST- 2013/10/23 06:00 [medline]
PHST- 2013/04/03 00:00 [pmc-release]
AID - PONE-D-12-21936 [pii]
AID - 10.1371/journal.pone.0060798 [doi]
PST - ppublish
SO  - PLoS One. 2013;8(4):e60798. doi: 10.1371/journal.pone.0060798. Epub 2013 Apr 3.