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WW Domain Containing Transcription Regulator regulates human conjunctiva epithelial cell proliferation via inhibiting TGFβ signaling pathway [corrected].

Tan XW, Beuerman RW, Poh CK, Mehta JS - Mol. Vis. (2012)

Bottom Line: Immortalized conjunctiva epithelial cells (NHC) were treated with TGFβ, targeting siRNA, TGFβ receptor antibody or TGFβ receptor inhibitor, to study the involvement of TAZ and TGFβ signaling pathway in conjunctiva cell proliferation by cell adhesion assay.Moreover, TGFβ receptor antibody and TGFβ receptor inhibitor rescued this anti-proliferative effect of TAZ siRNA.TAZ is involved in human conjunctiva epithelial cells proliferation via regulating TGFβ signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Singapore Eye Research Institute, Singapore.

ABSTRACT

Purpose: To investigate the role of Tafazzin (TAZ) protein in regulating the proliferation of normal human conjunctiva epithelial cells and epithelial cells from pterygium tissue.

Methods: Conjunctiva epithelial cells were cultured in keratinocytes growth medium and treated with transformation growth factor β (TGFβ) to analyze the expression and translocation of TAZ protein by immunostaining and BrdU analysis. Immortalized conjunctiva epithelial cells (NHC) were treated with TGFβ, targeting siRNA, TGFβ receptor antibody or TGFβ receptor inhibitor, to study the involvement of TAZ and TGFβ signaling pathway in conjunctiva cell proliferation by cell adhesion assay. Conjunctiva tissues from a normal human eye and an eye with pterygium disease were collected for histological analyses and western blot to evaluate the TAZ protein expression in vivo.

Results: TAZ expression was upregulated in mitotic conjunctiva epithelial cells, proliferating conjunctiva epithelial cells, TGFβ treated conjunctiva epithelial cells and human pterygium epithelium. TAZ siRNA induced less conjunctiva epithelial cell growth. Moreover, TGFβ receptor antibody and TGFβ receptor inhibitor rescued this anti-proliferative effect of TAZ siRNA.

Conclusions: TAZ is involved in human conjunctiva epithelial cells proliferation via regulating TGFβ signaling pathway.

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Related in: MedlinePlus

TAZ is responsive to TGFβ signal in conjunctiva epithelial cells. A: Fluorescent signals of TAZ protein (green) and smad4 (red) were upregulated after TGFβ treatment. Scale bar is 20 µm. B: Relative quantitation of the green fluorescent intensity of TAZ protein. Data were represented as mean±se, * p<0.01, ANOVA. C: Cell lysate of the epithelial cells with or without TGFβ treatment were blotted by anti-TAZ. Expression of TAZ protein was upregulated in TGFβ treated cells.
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f4: TAZ is responsive to TGFβ signal in conjunctiva epithelial cells. A: Fluorescent signals of TAZ protein (green) and smad4 (red) were upregulated after TGFβ treatment. Scale bar is 20 µm. B: Relative quantitation of the green fluorescent intensity of TAZ protein. Data were represented as mean±se, * p<0.01, ANOVA. C: Cell lysate of the epithelial cells with or without TGFβ treatment were blotted by anti-TAZ. Expression of TAZ protein was upregulated in TGFβ treated cells.

Mentions: It was recently reported that TAZ is responsive to TGFβ cytokine, forming a complex with Smad protein at the cytoplasm, which facilitated the nucleus translocation of Smad proteins in human embryonic stem cells [21]. In our study, we aimed to investigate whether the TAZ signal in human conjunctiva epithelial cells is also responsive to TGFβ. Our data showed that the fluorescence intensity of endogenous TAZ signal in epithelial cells that were treated with TGFβ is higher than that of cells without TGFβ treatment (Figure 4A). Smad4 staining was simultaneously performed as an indicator of the cells that were responsive to TGFβ. The relative quantitative fluorescence intensity in the nucleus of the epithelial cells was measured (Figure 4B). A total of 100 cells from three repeated experiments were analyzed. We found that after TGFβ treatment, fluorescent intensity of TAZ protein in the cell nucleus is about 2.5±0.3 times higher than that of untreated cells (Figure 4B). Western blot analysis further confirmed that the TAZ protein expression was upregulated in TGFβ treated cells (Figure 4C). Altogether, these results indicate that TAZ protein is responsive to TGFβ stimuli in human conjunctiva epithelial cells.


WW Domain Containing Transcription Regulator regulates human conjunctiva epithelial cell proliferation via inhibiting TGFβ signaling pathway [corrected].

Tan XW, Beuerman RW, Poh CK, Mehta JS - Mol. Vis. (2012)

TAZ is responsive to TGFβ signal in conjunctiva epithelial cells. A: Fluorescent signals of TAZ protein (green) and smad4 (red) were upregulated after TGFβ treatment. Scale bar is 20 µm. B: Relative quantitation of the green fluorescent intensity of TAZ protein. Data were represented as mean±se, * p<0.01, ANOVA. C: Cell lysate of the epithelial cells with or without TGFβ treatment were blotted by anti-TAZ. Expression of TAZ protein was upregulated in TGFβ treated cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3369895&req=5

f4: TAZ is responsive to TGFβ signal in conjunctiva epithelial cells. A: Fluorescent signals of TAZ protein (green) and smad4 (red) were upregulated after TGFβ treatment. Scale bar is 20 µm. B: Relative quantitation of the green fluorescent intensity of TAZ protein. Data were represented as mean±se, * p<0.01, ANOVA. C: Cell lysate of the epithelial cells with or without TGFβ treatment were blotted by anti-TAZ. Expression of TAZ protein was upregulated in TGFβ treated cells.
Mentions: It was recently reported that TAZ is responsive to TGFβ cytokine, forming a complex with Smad protein at the cytoplasm, which facilitated the nucleus translocation of Smad proteins in human embryonic stem cells [21]. In our study, we aimed to investigate whether the TAZ signal in human conjunctiva epithelial cells is also responsive to TGFβ. Our data showed that the fluorescence intensity of endogenous TAZ signal in epithelial cells that were treated with TGFβ is higher than that of cells without TGFβ treatment (Figure 4A). Smad4 staining was simultaneously performed as an indicator of the cells that were responsive to TGFβ. The relative quantitative fluorescence intensity in the nucleus of the epithelial cells was measured (Figure 4B). A total of 100 cells from three repeated experiments were analyzed. We found that after TGFβ treatment, fluorescent intensity of TAZ protein in the cell nucleus is about 2.5±0.3 times higher than that of untreated cells (Figure 4B). Western blot analysis further confirmed that the TAZ protein expression was upregulated in TGFβ treated cells (Figure 4C). Altogether, these results indicate that TAZ protein is responsive to TGFβ stimuli in human conjunctiva epithelial cells.

Bottom Line: Immortalized conjunctiva epithelial cells (NHC) were treated with TGFβ, targeting siRNA, TGFβ receptor antibody or TGFβ receptor inhibitor, to study the involvement of TAZ and TGFβ signaling pathway in conjunctiva cell proliferation by cell adhesion assay.Moreover, TGFβ receptor antibody and TGFβ receptor inhibitor rescued this anti-proliferative effect of TAZ siRNA.TAZ is involved in human conjunctiva epithelial cells proliferation via regulating TGFβ signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Singapore Eye Research Institute, Singapore.

ABSTRACT

Purpose: To investigate the role of Tafazzin (TAZ) protein in regulating the proliferation of normal human conjunctiva epithelial cells and epithelial cells from pterygium tissue.

Methods: Conjunctiva epithelial cells were cultured in keratinocytes growth medium and treated with transformation growth factor β (TGFβ) to analyze the expression and translocation of TAZ protein by immunostaining and BrdU analysis. Immortalized conjunctiva epithelial cells (NHC) were treated with TGFβ, targeting siRNA, TGFβ receptor antibody or TGFβ receptor inhibitor, to study the involvement of TAZ and TGFβ signaling pathway in conjunctiva cell proliferation by cell adhesion assay. Conjunctiva tissues from a normal human eye and an eye with pterygium disease were collected for histological analyses and western blot to evaluate the TAZ protein expression in vivo.

Results: TAZ expression was upregulated in mitotic conjunctiva epithelial cells, proliferating conjunctiva epithelial cells, TGFβ treated conjunctiva epithelial cells and human pterygium epithelium. TAZ siRNA induced less conjunctiva epithelial cell growth. Moreover, TGFβ receptor antibody and TGFβ receptor inhibitor rescued this anti-proliferative effect of TAZ siRNA.

Conclusions: TAZ is involved in human conjunctiva epithelial cells proliferation via regulating TGFβ signaling pathway.

Show MeSH
Related in: MedlinePlus