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Versican V1 Overexpression Induces a Myofibroblast-Like Phenotype in Cultured Fibroblasts.

Carthy JM, Meredith AJ, Boroomand S, Abraham T, Luo Z, Knight D, McManus BM - PLoS ONE (2015)

Bottom Line: These changes in cell function were associated with greater N-cadherin and integrin β1 expression, along with increased FAK phosphorylation.Consistent with this observation, the versican fibroblasts displayed increased synthetic activity, as measured by collagen III mRNA expression, as well as a greater capacity to contract a collagen lattice.Collectively, these data indicate versican expression induces a myofibroblast-like phenotype in cultured fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: UBC James Hogg Research Centre, Institute for Heart + Lung Health, Department of Pathology and Laboratory Medicine, University of British Columbia - Providence Health Care, Vancouver, British Columbia, Canada.

ABSTRACT

Background: Versican, a chondroitin sulphate proteoglycan, is one of the key components of the provisional extracellular matrix expressed after injury. The current study evaluated the hypothesis that a versican-rich matrix alters the phenotype of cultured fibroblasts.

Methods and results: The full-length cDNA for the V1 isoform of human versican was cloned and the recombinant proteoglycan was expressed in murine fibroblasts. Versican expression induced a marked change in fibroblast phenotype. Functionally, the versican-expressing fibroblasts proliferated faster and displayed enhanced cell adhesion, but migrated slower than control cells. These changes in cell function were associated with greater N-cadherin and integrin β1 expression, along with increased FAK phosphorylation. The versican-expressing fibroblasts also displayed expression of smooth muscle α-actin, a marker of myofibroblast differentiation. Consistent with this observation, the versican fibroblasts displayed increased synthetic activity, as measured by collagen III mRNA expression, as well as a greater capacity to contract a collagen lattice. These changes appear to be mediated, at least in part, by an increase in active TGF-β signaling in the versican expressing fibroblasts, and this was measured by phosphorylation and nuclear accumulation of SMAD2.

Conclusions: Collectively, these data indicate versican expression induces a myofibroblast-like phenotype in cultured fibroblasts.

No MeSH data available.


Related in: MedlinePlus

Versican alters the function of fibroblasts.(A) Versican increased cell proliferation 1.44 ± 0.07 fold over control cells, p<0.05. (B) Versican expressing fibroblasts showed decreased cell migration, as measure by in vitro scrape wound assay. (C) Cell adhesion was found to be increased in versican-transfected fibroblasts, measured at 30 minutes after seeding. (D) Representative Western blot showing increased integrin β1 expression, as well as phosphorylation of FAK-397, in versican-transfected fibroblasts. (* denotes p<0.05).
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pone.0133056.g003: Versican alters the function of fibroblasts.(A) Versican increased cell proliferation 1.44 ± 0.07 fold over control cells, p<0.05. (B) Versican expressing fibroblasts showed decreased cell migration, as measure by in vitro scrape wound assay. (C) Cell adhesion was found to be increased in versican-transfected fibroblasts, measured at 30 minutes after seeding. (D) Representative Western blot showing increased integrin β1 expression, as well as phosphorylation of FAK-397, in versican-transfected fibroblasts. (* denotes p<0.05).

Mentions: Although no major change in morphology was observed, the versican-expressing fibroblasts were found to function much differently than control cells. The versican fibroblasts proliferated significantly faster than control cells (Fig 3A, relative rate = 1.44 ± 0.07 of control cells, p<0.05). In contrast, the versican fibroblasts migrated slower than control cells (Fig 3B, 65 ±12 vs 96 ±3 percent wound closure, p<0.05). These cells also showed increased adhesion to culture dishes, as measured by cell spreading 30 minutes after seeding (Fig 3C, 3.43 ± 0.63 fold increase in cell surface area, p<0.05). Concomitant with increased cell adhesion, the versican fibroblasts displayed increased expression of integrin β1 as well as phosphorylation of focal adhesion kinase (FAK) (Fig 3D, 2.47 ± 0.41 and 2.01 ± 0.44 fold increase respectively, p<0.05).


Versican V1 Overexpression Induces a Myofibroblast-Like Phenotype in Cultured Fibroblasts.

Carthy JM, Meredith AJ, Boroomand S, Abraham T, Luo Z, Knight D, McManus BM - PLoS ONE (2015)

Versican alters the function of fibroblasts.(A) Versican increased cell proliferation 1.44 ± 0.07 fold over control cells, p<0.05. (B) Versican expressing fibroblasts showed decreased cell migration, as measure by in vitro scrape wound assay. (C) Cell adhesion was found to be increased in versican-transfected fibroblasts, measured at 30 minutes after seeding. (D) Representative Western blot showing increased integrin β1 expression, as well as phosphorylation of FAK-397, in versican-transfected fibroblasts. (* denotes p<0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4503433&req=5

pone.0133056.g003: Versican alters the function of fibroblasts.(A) Versican increased cell proliferation 1.44 ± 0.07 fold over control cells, p<0.05. (B) Versican expressing fibroblasts showed decreased cell migration, as measure by in vitro scrape wound assay. (C) Cell adhesion was found to be increased in versican-transfected fibroblasts, measured at 30 minutes after seeding. (D) Representative Western blot showing increased integrin β1 expression, as well as phosphorylation of FAK-397, in versican-transfected fibroblasts. (* denotes p<0.05).
Mentions: Although no major change in morphology was observed, the versican-expressing fibroblasts were found to function much differently than control cells. The versican fibroblasts proliferated significantly faster than control cells (Fig 3A, relative rate = 1.44 ± 0.07 of control cells, p<0.05). In contrast, the versican fibroblasts migrated slower than control cells (Fig 3B, 65 ±12 vs 96 ±3 percent wound closure, p<0.05). These cells also showed increased adhesion to culture dishes, as measured by cell spreading 30 minutes after seeding (Fig 3C, 3.43 ± 0.63 fold increase in cell surface area, p<0.05). Concomitant with increased cell adhesion, the versican fibroblasts displayed increased expression of integrin β1 as well as phosphorylation of focal adhesion kinase (FAK) (Fig 3D, 2.47 ± 0.41 and 2.01 ± 0.44 fold increase respectively, p<0.05).

Bottom Line: These changes in cell function were associated with greater N-cadherin and integrin β1 expression, along with increased FAK phosphorylation.Consistent with this observation, the versican fibroblasts displayed increased synthetic activity, as measured by collagen III mRNA expression, as well as a greater capacity to contract a collagen lattice.Collectively, these data indicate versican expression induces a myofibroblast-like phenotype in cultured fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: UBC James Hogg Research Centre, Institute for Heart + Lung Health, Department of Pathology and Laboratory Medicine, University of British Columbia - Providence Health Care, Vancouver, British Columbia, Canada.

ABSTRACT

Background: Versican, a chondroitin sulphate proteoglycan, is one of the key components of the provisional extracellular matrix expressed after injury. The current study evaluated the hypothesis that a versican-rich matrix alters the phenotype of cultured fibroblasts.

Methods and results: The full-length cDNA for the V1 isoform of human versican was cloned and the recombinant proteoglycan was expressed in murine fibroblasts. Versican expression induced a marked change in fibroblast phenotype. Functionally, the versican-expressing fibroblasts proliferated faster and displayed enhanced cell adhesion, but migrated slower than control cells. These changes in cell function were associated with greater N-cadherin and integrin β1 expression, along with increased FAK phosphorylation. The versican-expressing fibroblasts also displayed expression of smooth muscle α-actin, a marker of myofibroblast differentiation. Consistent with this observation, the versican fibroblasts displayed increased synthetic activity, as measured by collagen III mRNA expression, as well as a greater capacity to contract a collagen lattice. These changes appear to be mediated, at least in part, by an increase in active TGF-β signaling in the versican expressing fibroblasts, and this was measured by phosphorylation and nuclear accumulation of SMAD2.

Conclusions: Collectively, these data indicate versican expression induces a myofibroblast-like phenotype in cultured fibroblasts.

No MeSH data available.


Related in: MedlinePlus