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R ac1 modulates the vitreous‐induced plasticity of mesenchymal movement in retinal pigment epithelial cells
Author(s) -
Huang Xionggao,
Chen Youzhen,
Zhang Zhaotian,
Wei Yantao,
Ma Haizhi,
Zhang Ting,
Zhang Shaochong
Publication year - 2013
Publication title -
clinical and experimental ophthalmology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.3
H-Index - 74
eISSN - 1442-9071
pISSN - 1442-6404
DOI - 10.1111/ceo.12070
Subject(s) - microbiology and biotechnology , cell migration , retinal , retinal pigment epithelium , rho associated protein kinase , chemistry , biology , cell , phosphorylation , biochemistry
Background The vitreous has been shown to induce epithelial‐mesenchymal transdifferentiation because it induces fibroblast‐like morphology, enhanced migration and invasion in retinal pigment epithelial cells in proliferative vitreoretinopathy. R ac1 is the principal mediator of cell migration. In the current study, the relationship between R ac1 and cell migration, and invasion in vitreous‐transformed retinal pigment epithelial cells was investigated using NSC23766 , a specific inhibitor of R ac guanosine‐5′‐triphosphatase activity, and the involvement of a R ac1 guanosine‐5′‐triphosphatase‐dependent pathway was detected. Design One‐way design with multiple levels and repeated measurement design. Participants and Samples The vitreous humor was collected from 20 healthy donor eyes and the retinal pigment epithelial cells were obtained from 9 healthy donor eyes. Methods Human low‐passage retinal pigment epithelial cells were treated with normal medium or 25% vitreous medium. R ac1 activity was measured using a pull‐down assay. The cytotoxicity of NSC23766 was measured using the trypan blue dye exclusion test. Cell migration was measured using a wound healing assay. Cell invasion was determined using a transwell invasion assay. Protein expression of R ac1 and phosphorylation of LIM kinase 1 and cofilin were detected by W estern blot analysis. Main Outcome Measures Cell migration, invasion, Rac1 activity and phosphorylation of LIM kinase 1 and cofilin. Results R ac1guanosine‐5′‐triphosphatase was activated in vitreous‐transformed retinal pigment epithelial cells. A R ac inhibitor suppressed vitreous‐induced migration and invasion in retinal pigment epithelial cells. Cofilin phosphorylation was activated by vitreous treatment but blocked by NSC23766 . Conclusions R ac1 mediates vitreous‐transformed retinal pigment epithelial cells' plasticity of mesenchymal movement via R ac1 guanosine‐5′‐triphosphatase‐dependent pathways that modulate LIM kinase 1 and cofilin activity. R ac inhibition may be considered a novel treatment for proliferative vitreoretinopathy.