May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
TGF B1 and PMA Treatments Indirectly Regulate PAI-2 Expression in Corneal and Conjunctival Epithelial Cells
Author Affiliations & Notes
  • M. Massaro-Giordano
    Ophthalmology, Scheie Eye Institute, Philadelphia, Pennsylvania
  • M. Montanari
    Temple, College of Science and Technology, Sbarro Institute for Cancer Research and Molecular Medicine, Philadelphia, Pennsylvania
    Human Pathology and Oncology, University of Siena, Siena, Italy
  • C. M. Marshall
    Dermatology, University of Pennslyvania, Philadelphia, Pennsylvania
  • M. Macaluso
    Temple, College of Science and Technology, Sbarro Institute for Cancer Research and Molecular Medicine, Philadelphia, Pennsylvania
    Human Pathology and Oncology, University of Siena, Siena, Italy
  • Footnotes
    Commercial Relationships  M. Massaro-Giordano, None; M. Montanari, None; C.M. Marshall, None; M. Macaluso, None.
  • Footnotes
    Support  Sbarro Health Research Organization and in part by RPE
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 4311. doi:https://doi.org/
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    • Get Citation

      M. Massaro-Giordano, M. Montanari, C. M. Marshall, M. Macaluso; TGF B1 and PMA Treatments Indirectly Regulate PAI-2 Expression in Corneal and Conjunctival Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4311. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Several studies have reported that plaminogen activator inhibitor type 2 (PAI-2) is a multifunctional protein. It plays a role in cell differentiation, prevention of programmed cell death, regulation of cell proliferation, inhibition of microbial proteinases and in protection against stromal degradation.Previously we observed a different expression pattern of PAI-2 protein in the epithelium of the cornea and conjunctiva. We reported an interesting nuclear interaction between PAI-2 and the pRb2/130 proteins and suggested a model of how pRb2/p130 and PAI-2 may cooperate with specific chromatin remodeling enzymes in modulating PAI-2 transcription. We investigated the effects of transforming growth factor β-1 (TGF-β1) and phorbol 12-myristate 13-acetate (PMA) on the PAI-2 gene and protein expression and found that in both corneal and conjunctival cells TGF-β1 treatment down-regulates PAI-2 gene expression and decreases PAI-2 protein levels, while PMA treatment up-regulates the expression. Here we investigated the mechanisms by which TGF-β1 and PMA induce changes in both PAI-2 gene and protein expression in primary corneal and conjunctival cells. Our hypothesis is that following the treatments, specific signal transduction pathways may be turned on, which may affect pRb2/p130 and PAI-2 interaction and the binding on PAI-2 promoter.

Methods: : We assessed the binding of pRb2/p130 and PAI-2 protein to PAI-2 promoter by performing XChIP assay in the presence or absence of TGF-β1 and PMA. We assessed the interaction of pRb2/p130 and PAI-2 proteins by co-immunoprecipitation assay in the presence or absence of TGF-β1 and PMA.

Results: : We reported that TGF-β1 or PMA treatments do not affect the interaction of PAI-2 and pRb2/p130 on the PAI-2 proximal promoter. After the treatments these proteins still bind the PAI-2 promoter. We confirmed that after TGF-β1 treatment pRb2/p130 and PAI-2 proteins co-immunoprecipitate only in the nucleus of both corneal and conjunctival cells.

Keywords: cornea: epithelium • conjunctiva • gene/expression 
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