P311 induces the transdifferentiation of epidermal stem cells to myofibroblast-like cells by stimulating transforming growth factor β1 expression
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Author(s)
Type
Journal Article
Abstract
Background:
Epithelial to mesenchymal transition, especially to myofibroblasts, plays an important role in wound
healing, fibrosis, and carcinogenesis. Epidermal stem cells (EpSCs) are responsible for epidermal renewal and wound
re-epithelialization. However, it remains unclear whether and how EpSCs transdifferentiate into myofibroblasts or
myofibroblast-like cells (MFLCs). Here, we provide the first evidence showing that P311 induces EpSC to MFLC
transdifferentiation (EpMyT) via TGF
β
1/Smad signaling.
Methods:
Wound healing and mesenchymal features were observed in the P311 KO and P311 WT mouse model
of superficial second-degree burns. After the primary human or mouse EpSCs were forced to highly express P311
using an adenoviral vector, EpMyT was observed by immunofluorescence, real-time PCR, and western blot. The
activity of TGF
β
1 and Smad2/3 in EpSCs with different P311 levels was observed by western blot. The T
β
RI/II
inhibitor LY2109761 and Smad3 siRNA were applied to block the EpMyT in P311-overexpressing EpSCs and
exogenous TGF
β
1 was to restore the EpMyT in P311 KO EpSCs. Furthermore, the mechanism of P311 regulating
TGF
β
1 was investigated by bisulfite sequencing PCR, luciferase activity assay, and real-time PCR.
Results:
P311 KO mouse wounds showed delayed re-epithelialization and reduced mesenchymal features.
ThehumanormouseEpSCswithoverexpressedP311exhib
ited fusiform morphological changes, upregulated
expression of myofibroblast markers (
α
-SMA and vimentin), and downregu
lated expression of EpSC markers
(
β
1-integrin and E-cadherin). P311-expressing EpSCs showed decreased TGF
β
1mRNAandincreasedTGF
β
1
protein, T
β
RI/II mRNA, and activated Smad2/3. Moreover, LY2109761 and Smad3 siRNA reversed P311-induced
EpMyT. Under the stimulation of exogenous TGF
β
1, the phosphorylation of Smad2 and Smad3 in P311 KO EpSCs
was significantly lower than that in P311 WT EpSCs and the EpMyT in P311 KO EpSCs was restored. Furthermore,
P311 enhanced the methylation of TGF
β
1 promoter and increased activities of TGF
β
15
′
/3
′
untranslated regions
(UTRs) to stimulate TGF
β
1 expression. P311
+
α
-SMA
+
cells and P311
+
vimentin
+
cells were observed in the
epidermis of human burn wounds. Also, P311 was upregulated by IL-1
β
, IL-6, TNF
α
,andhypoxia.
Conclusions:
P311 is a novel TGF
β
1/Smad signaling-mediated regulator o
f transdifferentiation in EpSCs during
cutaneous wound healing. Furthermore, P311 might stimulate TGF
β
1 expression by promoting TGF
β
1 promoter
methylation and by activating the TGF
β
15
′
/3
′
UTR.
Epithelial to mesenchymal transition, especially to myofibroblasts, plays an important role in wound
healing, fibrosis, and carcinogenesis. Epidermal stem cells (EpSCs) are responsible for epidermal renewal and wound
re-epithelialization. However, it remains unclear whether and how EpSCs transdifferentiate into myofibroblasts or
myofibroblast-like cells (MFLCs). Here, we provide the first evidence showing that P311 induces EpSC to MFLC
transdifferentiation (EpMyT) via TGF
β
1/Smad signaling.
Methods:
Wound healing and mesenchymal features were observed in the P311 KO and P311 WT mouse model
of superficial second-degree burns. After the primary human or mouse EpSCs were forced to highly express P311
using an adenoviral vector, EpMyT was observed by immunofluorescence, real-time PCR, and western blot. The
activity of TGF
β
1 and Smad2/3 in EpSCs with different P311 levels was observed by western blot. The T
β
RI/II
inhibitor LY2109761 and Smad3 siRNA were applied to block the EpMyT in P311-overexpressing EpSCs and
exogenous TGF
β
1 was to restore the EpMyT in P311 KO EpSCs. Furthermore, the mechanism of P311 regulating
TGF
β
1 was investigated by bisulfite sequencing PCR, luciferase activity assay, and real-time PCR.
Results:
P311 KO mouse wounds showed delayed re-epithelialization and reduced mesenchymal features.
ThehumanormouseEpSCswithoverexpressedP311exhib
ited fusiform morphological changes, upregulated
expression of myofibroblast markers (
α
-SMA and vimentin), and downregu
lated expression of EpSC markers
(
β
1-integrin and E-cadherin). P311-expressing EpSCs showed decreased TGF
β
1mRNAandincreasedTGF
β
1
protein, T
β
RI/II mRNA, and activated Smad2/3. Moreover, LY2109761 and Smad3 siRNA reversed P311-induced
EpMyT. Under the stimulation of exogenous TGF
β
1, the phosphorylation of Smad2 and Smad3 in P311 KO EpSCs
was significantly lower than that in P311 WT EpSCs and the EpMyT in P311 KO EpSCs was restored. Furthermore,
P311 enhanced the methylation of TGF
β
1 promoter and increased activities of TGF
β
15
′
/3
′
untranslated regions
(UTRs) to stimulate TGF
β
1 expression. P311
+
α
-SMA
+
cells and P311
+
vimentin
+
cells were observed in the
epidermis of human burn wounds. Also, P311 was upregulated by IL-1
β
, IL-6, TNF
α
,andhypoxia.
Conclusions:
P311 is a novel TGF
β
1/Smad signaling-mediated regulator o
f transdifferentiation in EpSCs during
cutaneous wound healing. Furthermore, P311 might stimulate TGF
β
1 expression by promoting TGF
β
1 promoter
methylation and by activating the TGF
β
15
′
/3
′
UTR.
Date Issued
2016-12-01
Date Acceptance
2016-10-11
Citation
Stem Cell Research & Therapy, 2016, 7
ISSN
1757-6512
Publisher
BioMed Central
Journal / Book Title
Stem Cell Research & Therapy
Volume
7
Copyright Statement
© The Author(s). 2016
Open Access
This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Open Access
This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Subjects
Epidermal stem cells
Myofibroblast-like cells
P311
Transdifferentiation
Transforming growth factor beta 1
10 Technology
06 Biological Sciences
11 Medical And Health Sciences
Publication Status
Published
Article Number
175