Identification of transcriptional targets of Wnt/β-catenin signaling in dermal papilla cells of human scalp hair follicles: EP2 is a novel transcriptional target of Wnt3a

Abstract Background Recent studies showed that Wnt signaling through the β-catenin pathway (canonical Wnt signaling) act on mouse dermal papilla cells (DPCs) enabling hair follicles to keep growing. Objective To investigate whether human DPCs respond to canonical Wnt signaling and, if so, to identif...

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Bibliographic Details
Published in:Journal of dermatological science 2010-05, Vol.58 (2), p.91-96
Main Authors: Shin, HyeRim, Kwack, Mi Hee, Shin, Seung Hyun, Oh, Ji Won, Kang, Bo Mi, Kim, Ahnsup Andrew, Kim, Jinoh, Kim, Moon Kyu, Kim, Jung Chul, Sung, Young Kwan
Format: Article
Language:English
Subjects:
beta Catenin - metabolism
Cell Nucleus - metabolism
Cyclic AMP - metabolism
Dermal papilla
Dermatology
EP2
Hair follicle
Hair Follicle - metabolism
Humans
Immunoenzyme Techniques - methods
Microarray
Microscopy, Fluorescence - methods
Models, Biological
Oligonucleotide Array Sequence Analysis
PGE2
Receptors, Prostaglandin E - metabolism
Receptors, Prostaglandin E, EP2 Subtype
Reverse Transcriptase Polymerase Chain Reaction
Scalp - metabolism
Signal Transduction
Skin - metabolism
Wnt Proteins - metabolism
Wnt3 Protein
Wnt3a
Wnt3A Protein
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Summary:Abstract Background Recent studies showed that Wnt signaling through the β-catenin pathway (canonical Wnt signaling) act on mouse dermal papilla cells (DPCs) enabling hair follicles to keep growing. Objective To investigate whether human DPCs respond to canonical Wnt signaling and, if so, to identify target genes of Wnt/β-catenin pathway. Methods Cultured human DPCs were transiently transfected with the β-catenin responsive TCF reporter plasmid (pTopflash) and corresponding negative control reporter (pFopflash) to assess the activity of β-catenin signaling by Wnt3a (one of the canonical Wnts). Immunofluorescence staining was also performed to localize β-catenin in the presence or absence of Wnt3a. Microarray was carried out using Affymetrix gene chips. RT-PCR analysis and immunoblot were employed to verify microarray data. Cyclic AMP (cAMP) levels were measured using EIA assay after Wnt3a and PGE2 treatment in DPCs. Results Wnt3a significantly stimulated the transcriptional activity of pTopflash but not pFopflash. In line with this, we identified a number of genes that are regulated by Wnt3a. Some of the differently expressed genes including EP2 were confirmed by RT-PCR analysis. Immunoblot further confirmed that EP2 protein is indeed increased by Wnt3a. DPCs pretreated with Wnt3a showed higher responsiveness to PGE2 as measured by cAMP levels. Conclusions Elucidation of the role of Wnt3a-regulated genes identified in this study including EP2 would help our understanding of hair-induction and maintenance of anagen phase.
ISSN:0923-1811
1873-569X
DOI:10.1016/j.jdermsci.2010.02.011