7-ketocholesterol stimulates differentiation of lens epithelial cells

Detalhes bibliográficos
Autor(a) principal: Girão, Henrique
Data de Publicação: 2003
Outros Autores: Shang, Fu, Pereira, Paulo
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10316/12774
Resumo: PURPOSE: To establish if oxysterols stimulate differentiation of lens epithelial cells (LEC). METHODS: Primary cultures of lens epithelial cells were incubated with 7-ketocholesterol (7-keto), 25-hydroxycholesterol (25-OH) or cholesterol at 10 microg/ml for 10 days. Cells incubated with 100 ng/ml basic fibroblast growth factor (b-FGF) were used as positive controls for differentiation. The expression of the differentiation marker p57KIP2, proliferation marker PCNA (Proliferating Cell Nuclear Antigen) and fibers specific proteins gamma-crystallin, CP49, MIP26 following treatment with oxysterols was determined by western blot. Differentiation into fiber cells was further confirmed by counting the number of lentoid bodies formed following incubation with 7-keto. RESULTS: LEC incubated with 7-keto presented higher levels of p57KIP2 and showed expression of fiber specific proteins such as MIP26 and CP49, compared to cells incubated with 25-OH or cholesterol. The differentiation marker p57KIP2 increased over time for cells incubated with 7-keto while there was a decline on the amount of the proliferation marker PCNA. The expression of the fiber specific proteins gamma-crystallin, MIP26 and CP49 was detected after 5 days of incubation with 7-keto. Differentiation was accompanied by a seven-fold increase in the number of lentoid bodies formed. CONCLUSIONS: Results show for the first time that 7-keto inhibits proliferation and stimulates differentiation of lens epithelial cells into fiber cells. The presence of 7-keto in the lens may disrupt the highly regulated differentiation program of LEC, compromising normal lens growth and transparency
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spelling 7-ketocholesterol stimulates differentiation of lens epithelial cellsPURPOSE: To establish if oxysterols stimulate differentiation of lens epithelial cells (LEC). METHODS: Primary cultures of lens epithelial cells were incubated with 7-ketocholesterol (7-keto), 25-hydroxycholesterol (25-OH) or cholesterol at 10 microg/ml for 10 days. Cells incubated with 100 ng/ml basic fibroblast growth factor (b-FGF) were used as positive controls for differentiation. The expression of the differentiation marker p57KIP2, proliferation marker PCNA (Proliferating Cell Nuclear Antigen) and fibers specific proteins gamma-crystallin, CP49, MIP26 following treatment with oxysterols was determined by western blot. Differentiation into fiber cells was further confirmed by counting the number of lentoid bodies formed following incubation with 7-keto. RESULTS: LEC incubated with 7-keto presented higher levels of p57KIP2 and showed expression of fiber specific proteins such as MIP26 and CP49, compared to cells incubated with 25-OH or cholesterol. The differentiation marker p57KIP2 increased over time for cells incubated with 7-keto while there was a decline on the amount of the proliferation marker PCNA. The expression of the fiber specific proteins gamma-crystallin, MIP26 and CP49 was detected after 5 days of incubation with 7-keto. Differentiation was accompanied by a seven-fold increase in the number of lentoid bodies formed. CONCLUSIONS: Results show for the first time that 7-keto inhibits proliferation and stimulates differentiation of lens epithelial cells into fiber cells. The presence of 7-keto in the lens may disrupt the highly regulated differentiation program of LEC, compromising normal lens growth and transparencyMolecular Vision2003-10-06info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/12774http://hdl.handle.net/10316/12774engMolecular Vision. 9 (2003) 497-5011090-0535Girão, HenriqueShang, FuPereira, Pauloinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2022-08-23T14:28:43Zoai:estudogeral.uc.pt:10316/12774Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:43:37.985668Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv 7-ketocholesterol stimulates differentiation of lens epithelial cells
title 7-ketocholesterol stimulates differentiation of lens epithelial cells
spellingShingle 7-ketocholesterol stimulates differentiation of lens epithelial cells
Girão, Henrique
title_short 7-ketocholesterol stimulates differentiation of lens epithelial cells
title_full 7-ketocholesterol stimulates differentiation of lens epithelial cells
title_fullStr 7-ketocholesterol stimulates differentiation of lens epithelial cells
title_full_unstemmed 7-ketocholesterol stimulates differentiation of lens epithelial cells
title_sort 7-ketocholesterol stimulates differentiation of lens epithelial cells
author Girão, Henrique
author_facet Girão, Henrique
Shang, Fu
Pereira, Paulo
author_role author
author2 Shang, Fu
Pereira, Paulo
author2_role author
author
dc.contributor.author.fl_str_mv Girão, Henrique
Shang, Fu
Pereira, Paulo
description PURPOSE: To establish if oxysterols stimulate differentiation of lens epithelial cells (LEC). METHODS: Primary cultures of lens epithelial cells were incubated with 7-ketocholesterol (7-keto), 25-hydroxycholesterol (25-OH) or cholesterol at 10 microg/ml for 10 days. Cells incubated with 100 ng/ml basic fibroblast growth factor (b-FGF) were used as positive controls for differentiation. The expression of the differentiation marker p57KIP2, proliferation marker PCNA (Proliferating Cell Nuclear Antigen) and fibers specific proteins gamma-crystallin, CP49, MIP26 following treatment with oxysterols was determined by western blot. Differentiation into fiber cells was further confirmed by counting the number of lentoid bodies formed following incubation with 7-keto. RESULTS: LEC incubated with 7-keto presented higher levels of p57KIP2 and showed expression of fiber specific proteins such as MIP26 and CP49, compared to cells incubated with 25-OH or cholesterol. The differentiation marker p57KIP2 increased over time for cells incubated with 7-keto while there was a decline on the amount of the proliferation marker PCNA. The expression of the fiber specific proteins gamma-crystallin, MIP26 and CP49 was detected after 5 days of incubation with 7-keto. Differentiation was accompanied by a seven-fold increase in the number of lentoid bodies formed. CONCLUSIONS: Results show for the first time that 7-keto inhibits proliferation and stimulates differentiation of lens epithelial cells into fiber cells. The presence of 7-keto in the lens may disrupt the highly regulated differentiation program of LEC, compromising normal lens growth and transparency
publishDate 2003
dc.date.none.fl_str_mv 2003-10-06
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10316/12774
http://hdl.handle.net/10316/12774
url http://hdl.handle.net/10316/12774
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Molecular Vision. 9 (2003) 497-501
1090-0535
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dc.publisher.none.fl_str_mv Molecular Vision
publisher.none.fl_str_mv Molecular Vision
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