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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 protein clusters at the nucleus of proliferating epithelial cells. Conjunctiva epithelial cells were labeled for BrdU (red) and TAZ (green), DAPI was applied as nucleus counter staining dye (blue). White arrows indicate a BrdU positive epithelial cell also stained by high intensity of TAZ protein. Scale bar is 20 µm.
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f3: TAZ protein clusters at the nucleus of proliferating epithelial cells. Conjunctiva epithelial cells were labeled for BrdU (red) and TAZ (green), DAPI was applied as nucleus counter staining dye (blue). White arrows indicate a BrdU positive epithelial cell also stained by high intensity of TAZ protein. Scale bar is 20 µm.

Mentions: BrdU analysis was performed to further examine whether TAZ protein is upregulated in proliferating conjunctiva epithelial cells. We observed that in BrdU positive cells, TAZ protein was found clustering in the nucleus of the cells, while in cells that were BrdU negative, TAZ protein was predominantly expressed in the cytoplasm (Figure 3). Moreover, the fluorescence intensity of TAZ signal in BrdU positive cells was also observed to be much higher than that of BrdU negative cells. This result is consistent with our previous observation that TAZ protein was highly upregulated during epithelial cell division and proliferation; therefore, the activation and translocation of TAZ protein are closely associated with epithelial cell proliferation.


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 protein clusters at the nucleus of proliferating epithelial cells. Conjunctiva epithelial cells were labeled for BrdU (red) and TAZ (green), DAPI was applied as nucleus counter staining dye (blue). White arrows indicate a BrdU positive epithelial cell also stained by high intensity of TAZ protein. Scale bar is 20 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: TAZ protein clusters at the nucleus of proliferating epithelial cells. Conjunctiva epithelial cells were labeled for BrdU (red) and TAZ (green), DAPI was applied as nucleus counter staining dye (blue). White arrows indicate a BrdU positive epithelial cell also stained by high intensity of TAZ protein. Scale bar is 20 µm.
Mentions: BrdU analysis was performed to further examine whether TAZ protein is upregulated in proliferating conjunctiva epithelial cells. We observed that in BrdU positive cells, TAZ protein was found clustering in the nucleus of the cells, while in cells that were BrdU negative, TAZ protein was predominantly expressed in the cytoplasm (Figure 3). Moreover, the fluorescence intensity of TAZ signal in BrdU positive cells was also observed to be much higher than that of BrdU negative cells. This result is consistent with our previous observation that TAZ protein was highly upregulated during epithelial cell division and proliferation; therefore, the activation and translocation of TAZ protein are closely associated with epithelial cell proliferation.

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