Reference - Detail
RRC ID | 43216 |
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Author | Ko JA, Liu Y, Yanai R, Chikama T, Takezawa T, Nishida T. |
Title | Upregulation of tight-junctional proteins in corneal epithelial cells by corneal fibroblasts in collagen vitrigel cultures. |
Journal | Invest Ophthalmol Vis Sci |
Abstract |
PURPOSE:To investigate the effects of corneal fibroblasts on the differentiation of corneal epithelial cells in a coculture system based on a collagen vitrigel membrane. METHODS:Simian virus 40-transformed human corneal epithelial (HCE) cells and human corneal fibroblasts were cultured on opposite sides of a collagen vitrigel membrane. The distribution of HCE cells and corneal fibroblasts on the collagen membrane was determined by immunofluorescence staining and immunoblot analysis of marker proteins. Expression of the tight-junctional proteins ZO-1, occludin, and claudin and of the adherens-junctional proteins E- and N-cadherin in HCE cells was determined at the mRNA and protein levels by reverse transcription-polymerase chain reaction analysis and immunoblot analysis, respectively. RESULTS:The abundance of ZO-1, occludin, and claudin mRNA and proteins in HCE cells was markedly increased by coculture with corneal fibroblasts. The expression of E- or N-cadherin did not differ between HCE cells cultured with corneal fibroblasts and those cultured without them. PD98059, a specific inhibitor of signaling by extracellular signal regulated kinase (ERK), prevented the upregulation of tight-junctional proteins in HCE cells by corneal fibroblasts. CONCLUSIONS:Human corneal fibroblasts regulated the expression of tight-junctional proteins in HCE cells, suggesting that corneal fibroblasts may play an important role in the differentiation of corneal epithelial cells. |
Volume | 49(1) |
Pages | 113-9 |
Published | 2008-1-1 |
DOI | 10.1167/iovs.07-0353 |
PII | 49/1/113 |
PMID | 18172082 |
MeSH | Antigens, CD / genetics Antigens, CD / metabolism Cadherins / genetics Cadherins / metabolism Calcium-Calmodulin-Dependent Protein Kinases / pharmacology Cell Differentiation Cell Line, Transformed Claudin-1 Coculture Techniques Collagen Cornea / cytology Epithelium, Corneal / drug effects Epithelium, Corneal / metabolism* Fibroblasts / physiology* Flavonoids / pharmacology Fluorescent Antibody Technique, Indirect Humans Immunoblotting Membrane Proteins / genetics Membrane Proteins / metabolism* Occludin Phosphoproteins / genetics Phosphoproteins / metabolism* RNA, Messenger / metabolism Reverse Transcriptase Polymerase Chain Reaction Simian virus 40 Up-Regulation / physiology* Zonula Occludens-1 Protein |
IF | 3.47 |
Times Cited | 23 |
WOS Category | OPHTHALMOLOGY |
Resource | |
Human and Animal Cells | HCE-T(RCB2280) |