Abstract | PURPOSE: METHODS: Cultured human RPE cells were treated with 1.0 ng/mL TGF-beta2 for 24 hours. Cell surface tTG expression was determined by cell fraction analysis. Attachment on Col I, full-length Fn, and its 45-kDa gelatin-binding and 110-kDa cell-binding fragment was measured with an MTT assay. Migration of RPE cells was measured by a Boyden chamber assay, and cell spreading was determined. Experiments were performed in the presence or absence of anti-tTG antibodies and anti- integrin alpha5 and beta1 antibodies. RESULTS:
TGF-beta2 markedly induced expression of cell-surface tTG on RPE cells and increased attachment and migration on Fn and Col I. Blocking cell surface tTG inhibited attachment, migration, and spreading on Fn and its 45-kDa gelatin-binding fragment, whereas no effect was seen on Col I and the 110-kDa cell-binding Fn fragment. In contrast, blocking of integrin alpha5 and beta1 suppressed adhesion and migration on full-length Fn and the 110-kDa Fn fragment. CONCLUSIONS: These data demonstrate that TGF-beta2 increases expression of cell surface tTG, which in turn strengthens adhesion, migration, and spreading of RPE cells on Fn through the 45-kDa gelatin-binding Fn fragment. At the onset of PVR, this mechanism may help RPE cells to attach and migrate on Fn-containing matrices.
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Authors | Siegfried G Priglinger, Claudia S Alge, Aljoscha S Neubauer, Nadine Kristin, Christoph Hirneiss, Kirsten Eibl, Anselm Kampik, Ulrich Welge-Lussen |
Journal | Investigative ophthalmology & visual science
(Invest Ophthalmol Vis Sci)
Vol. 45
Issue 3
Pg. 955-63
(Mar 2004)
ISSN: 0146-0404 [Print] United States |
PMID | 14985317
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Collagen Type I
- Fibronectins
- Membrane Proteins
- TGFB2 protein, human
- Transforming Growth Factor beta
- Transforming Growth Factor beta2
- Gelatin
- Transglutaminases
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Topics |
- Adolescent
- Adult
- Aged
- Binding Sites
- Blotting, Western
- Cell Adhesion
(physiology)
- Cell Membrane
(drug effects)
- Cell Movement
(physiology)
- Cells, Cultured
- Collagen Type I
(metabolism)
- Fibronectins
(metabolism)
- Gelatin
(metabolism)
- Humans
- Membrane Proteins
(metabolism)
- Middle Aged
- Pigment Epithelium of Eye
(cytology, drug effects, enzymology)
- Protein Binding
- Transforming Growth Factor beta
(pharmacology)
- Transforming Growth Factor beta2
- Transglutaminases
(metabolism)
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