Epithelial-mesenchymal transition and proliferation of retinal pigment epithelial cells initiated upon loss of cell-cell contact.
OBJECTIVE
Molecular mechanisms that initiate epithelial-mesenchymal transition (EMT) involved in ocular fibrotic complications remain elusive. Studies were conducted to examine the role of cell-cell contact in regulating EMT and proliferation of retinal pigment epithelial (RPE) cells.


METHODS
Porcine RPE cells were isolated as sheets and cultured in vitro on lens capsules. Cell morphology was examined by microscopy. Western blot analysis and immunostaining were used to follow protein expression. Cell proliferation and RPE function were assessed by BrdU incorporation and phagocytosis assay, respectively.


RESULTS
RPE cells in the center of each sheet maintained cell-cell contacts and retained a differentiated phenotype. Disruption of cadherin function in these cells resulted in the loss of cell-cell contact and the concomitant induction of mesenchymal marker protein expression and cell proliferation. RPE cells at the edge of the sheet migrated away from the sheet, underwent EMT, and initiated proliferation, which was accompanied by a switch in cadherin expression from P-cadherin to N-cadherin. Although TGF-beta is thought to be a classic inducer of EMT, it was unable to initiate EMT in RPE cells maintaining cell-cell contact. However, change to alpha-SMA-positive myofibroblasts was induced by TGF-beta in cells that had already undergone EMT.


CONCLUSIONS
EMT and the onset of proliferation in RPE cells is initiated by loss of cell-cell contact. TGF-beta cannot initiate EMT or the proliferation of RPE cells maintaining cell-cell contact but appears to play an important secondary role downstream of EMT in inducing transition to a myofibroblast phenotype-a phenotype linked to the development of fibrotic complications.
Tamiya
Shigeo
S
Department of Ophthalmology and Visual Sciences, School of Medicine, University of Louisville, Louisville, KY 40202, USA. shigeo.tamiya@louisville.edu
Liu
LanHsin
L
Kaplan
Henry J
HJ
D000199:Actins / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology*; D002450:Cell Communication / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005136:Eye Proteins; D019084:Fluorescent Antibody Technique, Indirect; D008855:Microscopy, Electron, Scanning; D010587:Phagocytosis; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000378:metabolism; D012374:Rod Cell Outer Segment; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013552:Swine; D014746:Vimentin / Q000378:metabolism
D000199:Actins; D015820:Cadherins; D005136:Eye Proteins; D014746:Vimentin; D000254:Sodium-Potassium-Exchanging ATPase