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Chatri Ngamkitidechakul, Janice M. Burke, William J. O’Brien, Sally S. Twining; Maspin: Synthesis by Human Cornea and Regulation of In Vitro Stromal Cell Adhesion to Extracellular Matrix. Invest. Ophthalmol. Vis. Sci. 2001;42(13):3135-3141.
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purpose. Maspin, a tumor-suppressor protein that regulates cell migration,
invasion, and adhesion, is synthesized by many normal epithelial cells,
but downregulated in invasive epithelial tumor cells. The purpose of
this study was to determine whether cells in the normal human cornea
express maspin and whether maspin affects corneal stromal cell adhesion
to extracellular matrix molecules.
methods. Maspin expression was analyzed by immunodot blot, Western blot, and
RT-PCR analyses in cells obtained directly from human corneas in situ.
Maspin protein and mRNA were also studied in primary and passaged
cultures of corneal stromal cells using Western blot analysis, RT-PCR,
and immunofluorescence microscopy. Maspin cDNA was cloned and sequenced
from human corneal epithelial cells and expressed in a yeast system.
The recombinant maspin was used to study attachment of cultured human
corneal stromal cells to extracellular matrices.
results. Maspin mRNA and micromolar amounts of the protein were found in all
three layers of the human cornea in situ, including the stroma. Maspin
was also detected in primary and first-passage corneal stromal cells,
but its expression was downregulated in subsequent passages.
Late-passage stromal cells, which did not produce maspin, responded to
exogenous recombinant maspin as measured by increased cell adhesion not
only to fibronectin, similar to mammary gland tumor epithelial cells,
but also to type I collagen, type IV collagen, and laminin.
conclusions. The corneal stromal cell is the first nonepithelial cell type shown to
synthesize maspin. Loss of maspin expression in late-passage corneal
stromal cells in culture and their biological response to exogenous
maspin suggests a role for maspin on the stromal cells in the cornea.
Maspin may function within the cornea to regulate cell adhesion to
extracellular matrix molecules and perhaps to regulate the migration of
activated fibroblasts during corneal stromal wound
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