We conclude that PI3K-mediated GSK3ß phosphorylation is involved in the intestinal epithelial wound-healing response. Phosphorylation of GSK3ß may be important for intestinal restitution by promoting cell motility in response to wounding

A Dignass

A Sturm

Adrian John Harwood

AU Dignass

BK Dieckgraefe

Brigitte Allard

C Ciacci

C Jobin

CD Packey

Christian Jobin

CR Owen

E Beurel

EA Severson

ES Harris

HM Zhang

I Vivanco

IM Shih

J Rak

JA Engelman

JM Russo

K Myant

KA Steinbrecher

KP Hoeflich

L Koivisto

LJ Egan

M Bianchi

M Goke

M Kapoor

M Martin

MR Frey

ON El-Assal

P Cohen

P Nava

R Kajino-Sakamoto

R Mayr

RB Sartor

RE Bachelder

RJ Vaidya

S Basuroy

S Etienne-Manneville

SA McCormack

T Karrasch

T Sun

Tanja Spaeth

TC He

TF Bullen

Thomas Karrasch

WM de Araujo

WS Garrett

Y Takada

PLoS ONE

English

PubMed

The ability of the intestinal epithelial barrier to respond to various injurious insults is an essential component of intestinal homeostasis. However, the molecular mechanisms responsible for wound-healing and repair in the intestine are poorly understood. The glycogen synthase kinase 3ß (GSK3ß) has been implicated in various biological processes such as cellular motility, cell spreading and recently inflammation.To investigate the role of GSK3ß in intestinal epithelial cell restitution.Rat intestinal epithelial IEC18 cells were serum-starved for 16 to 24 h and wounded by multiple scraping. Akt(Ser473)-, GSK3ß(Ser9)- and RelA(Ser536)-phosphorylation were determined by Western blot using specific phospho-antibodies. The inhibitors AG1478 (1 µM) and Ly294002 (25 µM) were used to block EGF-R autophosphorylation and PI3K-activation, respectively. ß-Catenin/LEF/TCF dependent transcription was determined by reporter gene assay (TOP/FOP system). C-myc gene expression was evaluated by real-time RT-PCR. GSK3ß(-/-) mouse embryonic fibroblasts were used to characterize the role of GSK3ß in wounding-induced cell migration.Wounding induced GSK3ß(Ser9) phosphorylation in IEC-18 cells, which led to ß-catenin accumulation as well as nuclear translocation of ß-catenin. ß-Catenin stabilization/nuclear translocation led to enhanced LEF-TCF transcriptional activity and subsequent c-myc mRNA accumulation in wounded cell monolayers. Blocking PI3K/Akt signaling with Ly294002 prevented wound-induced GSK3ß(Ser9) phosphorylation as well as ß-catenin nuclear translocation and significantly attenuated restitution. Additionally, wounding induced rapid NF-kB(Ser536) phosphorylation, which was inhibited by AG1478, but not by Ly294002. GSK3ß(-/-) cells demonstrated significantly attenuated wound-induced restitution compared to wild-type cells.We conclude that PI3K-mediated GSK3ß phosphorylation is involved in the intestinal epithelial wound-healing response. Phosphorylation of GSK3ß may be important for intestinal restitution by promoting cell motility in response to wounding

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PI3K-dependent GSK3ß(Ser9)-phosphorylation is implicated in the intestinal epithelial cell wound-healing response.

Crossref

PI3K-Dependent GSK3ß(Ser9)-Phosphorylation Is Implicated in the Intestinal Epithelial Cell Wound-Healing Response

The ability of the intestinal epithelial barrier to respond to various injurious insults is an essential component of intestinal homeostasis. However, the molecular mechanisms responsible for wound-healing and repair in the intestine are poorly understood. The glycogen synthase kinase 3ß (GSK3ß) has been implicated in various biological processes such as cellular motility, cell spreading and recently inflammation.To investigate the role of GSK3ß in intestinal epithelial cell restitution. mouse embryonic fibroblasts were used to characterize the role of GSK3ß in wounding-induced cell migration. cells demonstrated significantly attenuated wound-induced restitution compared to wild-type cells.We conclude that PI3K-mediated GSK3ß phosphorylation is involved in the intestinal epithelial wound-healing response. Phosphorylation of GSK3ß may be important for intestinal restitution by promoting cell motility in response to wounding

Karrasch Thomas

Spaeth Tanja

Allard Brigitte

Jobin Christian

Public Library of Science (PLOS)

Introduction: The ability of the intestinal epithelial barrier to respond to various injurious insults is an essential component of intestinal homeostasis. However, the molecular mechanisms responsible for wound-healing and repair in the intestine are poorly understood. The glycogen synthase kinase 3 beta (GSK3 beta) has been implicated in various biological processes such as cellular motility, cell spreading and recently inflammation. Aim: To investigate the role of GSK3 beta in intestinal epithelial cell restitution. Methods: Rat intestinal epithelial IEC18 cells were serum-starved for 16 to 24h and wounded by multiple scraping. Akt(Ser473)-, GSK3 beta(Ser9)-and RelA(Ser536)-phosphorylation were determined by Western blot using specific phospho-antibodies. The inhibitors AG1478 (1 mu M) and Ly294002 (25 mM) were used to block EGF-R autophosphorylation and PI3K-activation, respectively. beta-catenin/LEF/TCF dependent transcription was determined by reporter gene assay (TOP/FOP system). C-myc gene expression was evaluated by real-time RT-PCR. GSK3 beta(-/-) mouse embryonic fibroblasts were used to characterize the role of GSK3 beta in wounding-induced cell migration. Results: Wounding induced GSK3 beta(Ser9) phosphorylation in IEC-18 cells, which led to beta-catenin accumulation as well as nuclear translocation of beta-catenin. beta-catenin stabilization/nuclear translocation led to enhanced LEF-TCF transcriptional activity and subsequent c-myc mRNA accumulation in wounded cell monolayers. Blocking PI3K/Akt signaling with Ly294002 prevented wound-induced GSK3 beta(Ser9) phosphorylation as well as beta-catenin nuclear translocation and significantly attenuated restitution. Additionally, wounding induced rapid NF-kB(Ser536) phosphorylation, which was inhibited by AG1478, but not by Ly294002. GSK3 beta(-/-) cells demonstrated significantly attenuated wound-induced restitution compared to wild-type cells. Conclusion: We conclude that PI3K-mediated GSK3 beta phosphorylation is involved in the intestinal epithelial wound-healing response. Phosphorylation of GSK3 beta may be important for intestinal restitution by promoting cell motility in response to wounding

PI3K-dependent GSK3ß(Ser9)-phosphorylation is implicated in the intestinal epithelial cell wound-healing response.

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