Senescence-associated β-galactosidase activity in the in vitro ovarian stromal fibroblasts

Introduction: A growing biological research field is the cellular senescence, a mechanism that has been associated, under certain circumstances, with malignant transformation. Given the high incidence of ovarian cancer and its main origin from the ovarian surface epithelium, as well as the possibili...

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Autores Principales: Chuaire-Noack, Lilian, García-Morcote, Cristian, Ramírez-Clavijo, Sandra
Formato: Artículo (Article)
Lenguaje:Español (Spanish)
Publicado: Universidad del Rosario 2011
Materias:
Acceso en línea:http://revistas.urosario.edu.co/index.php/revsalud/article/view/1545
http://repository.urosario.edu.co/handle/10336/7396
id ir-10336-7396
recordtype dspace
institution EdocUR - Universidad del Rosario
collection DSpace
language Español (Spanish)
topic β-galactosidase, replicative senescence, ovarían fibroblasts, epithelial-mesenchimal
β-galactosidasa, senescencia replicativa, fibroblastos ováricos, conversión epiteliomesenquimática
β-galactosidase, senescência replicativas, fibroblastos ovarianos, conversão epitéliomesenquimática
spellingShingle β-galactosidase, replicative senescence, ovarían fibroblasts, epithelial-mesenchimal
β-galactosidasa, senescencia replicativa, fibroblastos ováricos, conversión epiteliomesenquimática
β-galactosidase, senescência replicativas, fibroblastos ovarianos, conversão epitéliomesenquimática
Chuaire-Noack, Lilian
García-Morcote, Cristian
Ramírez-Clavijo, Sandra
Senescence-associated β-galactosidase activity in the in vitro ovarian stromal fibroblasts
description Introduction: A growing biological research field is the cellular senescence, a mechanism that has been associated, under certain circumstances, with malignant transformation. Given the high incidence of ovarian cancer and its main origin from the ovarian surface epithelium, as well as the possibility that an epithelial-mesenchymal transition occurs, we evaluated both the in vitro growth of stromal fibroblasts from the ovarian cortex and their β-galactosidase activity at pH 6, enzyme whose expression is considered as a marker of replicative senescence. Methods: 48 samples of ovarian cortical fibroblasts from donors without a history of cancer were serially cultured until the end of their replicative life. β-galactosidase activity at pH 6 was quantified in each passage by the chemiluminiscent method. As control, we used ovarian epithelial cell cultures from the same donors. The enzyme activity was also evaluated in fibroblasts previously induced to senescence by exposure to hydrogen peroxide. Results: The analysis of the enzyme activity and the replicative capacity taken together showed that the fibroblast cultures reached the senescent state at passages 4-5, as what happened with the control epithelial cells. Fibroblasts induced to senescence showed high variability in the values of enzymatic activity. Conclusions: The similarity between both types of cells in reaching the senescent state deserves to be taken into account in relation to the epithelialmesenchymal transition that has been proposed to explain their behavior in the genesis of cancer arising from ovarian surface epithelium. Low β-galactosidase activity values at pH 6 would suggest possible inactivation of the response pathways to oxidative stress.
format Artículo (Article)
author Chuaire-Noack, Lilian
García-Morcote, Cristian
Ramírez-Clavijo, Sandra
author_facet Chuaire-Noack, Lilian
García-Morcote, Cristian
Ramírez-Clavijo, Sandra
author_sort Chuaire-Noack, Lilian
title Senescence-associated β-galactosidase activity in the in vitro ovarian stromal fibroblasts
title_short Senescence-associated β-galactosidase activity in the in vitro ovarian stromal fibroblasts
title_full Senescence-associated β-galactosidase activity in the in vitro ovarian stromal fibroblasts
title_fullStr Senescence-associated β-galactosidase activity in the in vitro ovarian stromal fibroblasts
title_full_unstemmed Senescence-associated β-galactosidase activity in the in vitro ovarian stromal fibroblasts
title_sort senescence-associated β-galactosidase activity in the in vitro ovarian stromal fibroblasts
publisher Universidad del Rosario
publishDate 2011
url http://revistas.urosario.edu.co/index.php/revsalud/article/view/1545
http://repository.urosario.edu.co/handle/10336/7396
_version_ 1645140850824445952
spelling ir-10336-73962019-09-19T12:37:01Z Senescence-associated β-galactosidase activity in the in vitro ovarian stromal fibroblasts Actividad β-galactosidasa asociada con la senescencia en fibroblastos del estroma ovárico in vitro Atividade β-galactosidase associada com a senescência em fibroblastos do estroma ovariano in vitro Chuaire-Noack, Lilian García-Morcote, Cristian Ramírez-Clavijo, Sandra β-galactosidase, replicative senescence, ovarían fibroblasts, epithelial-mesenchimal β-galactosidasa, senescencia replicativa, fibroblastos ováricos, conversión epiteliomesenquimática β-galactosidase, senescência replicativas, fibroblastos ovarianos, conversão epitéliomesenquimática Introduction: A growing biological research field is the cellular senescence, a mechanism that has been associated, under certain circumstances, with malignant transformation. Given the high incidence of ovarian cancer and its main origin from the ovarian surface epithelium, as well as the possibility that an epithelial-mesenchymal transition occurs, we evaluated both the in vitro growth of stromal fibroblasts from the ovarian cortex and their β-galactosidase activity at pH 6, enzyme whose expression is considered as a marker of replicative senescence. Methods: 48 samples of ovarian cortical fibroblasts from donors without a history of cancer were serially cultured until the end of their replicative life. β-galactosidase activity at pH 6 was quantified in each passage by the chemiluminiscent method. As control, we used ovarian epithelial cell cultures from the same donors. The enzyme activity was also evaluated in fibroblasts previously induced to senescence by exposure to hydrogen peroxide. Results: The analysis of the enzyme activity and the replicative capacity taken together showed that the fibroblast cultures reached the senescent state at passages 4-5, as what happened with the control epithelial cells. Fibroblasts induced to senescence showed high variability in the values of enzymatic activity. Conclusions: The similarity between both types of cells in reaching the senescent state deserves to be taken into account in relation to the epithelialmesenchymal transition that has been proposed to explain their behavior in the genesis of cancer arising from ovarian surface epithelium. Low β-galactosidase activity values at pH 6 would suggest possible inactivation of the response pathways to oxidative stress. Introducción: Un campo de investigación creciente de la biología es la senescencia celular, mecanismo que ha sido asociado -bajo determinadas circunstancias- con la transformación maligna.Teniendo en cuenta la elevada incidencia de cáncer ovárico y su génesis preferencial a partir del epitelio superficial del ovario, así como la posibilidad de ocurrencia de una transición epiteliomesenquimática, se evaluó, tanto el crecimiento in vitro de los fibroblastos del estroma cortical, como la actividad a pH 6 de la β-galactosidasa, enzima cuya expresión ha sido clásicamente considerada como marcador de senescencia replicativa. Metodología: 48 muestras de fibroblastos de la corteza ovárica provenientes de donantes sin antecedentes de cáncer fueron cultivadas en forma seriada hasta el final de su vida replicativa. Mediante el método quimioluminiscente, en cada pase fue cuantificada la actividad β-galactosidasa a pH 6. Como control se utilizaron cultivos de células del epitelio superficial ovàrico de las mismas donantes. La actividad enzimática fue también evaluada en fibroblastos previamente inducidos a senescencia con peróxido de hidrógeno. Resultados: Las lecturas de actividad enzimática, analizadas en conjunto con la capacidad replicativa, indican que los cultivos de fibroblastos alcanzaron el estado senescente hacia los pases 4-5, lo que también ocurrió con las células epiteliales. Los fibroblastos inducidos a senescencia mostraron valores variables de actividad enzimática. Conclusiones: La semejanza entre los fibroblastos y las células epiteliales en cuanto al inicio de la senescencia podría estar relacionado con la transición epitelio-mesenquimática que ha sido descrita como factor de riesgo de cáncer derivado del epitelio superficial ovàrico. Valores bajos de actividad β-galactosidasa podrían sugerir que, en algunos casos, ocurrió inactivación de las vías de respuesta al estrés oxidativo. Introdução: um campo de pesquisa crescente da biologia é a senescência celular, mecanismo que tem sido associado, sob determinadas circunstancias, com a transformação maligna. Tendo em conta a elevada incidência de câncer ovariano e sua gênese preferencial a partir do epitélio superficial do ovário, assim como a possibilidade de ocorrência de uma transição epitélio-mesenquimática, se avaliou, tanto o crescimento in vitro dos fibroblastos do estroma cortical, como a atividade a pH 6 da β-galactosidase, enzima cuja expressão tem sido classicamente considerada como marcador de senescência replicativa. Metodologia: 48 amostras de fibroblastos do córtex ovariano provenientes de doadores sem antecedentes de câncer foram cultivadas em forma seriada até o final de sua vida replicativa. Mediante o método quimioluminescente, em cada passe foi quantificada a atividade β-galactosidase a pH 6. Como controle utilizou-se cultivos de células do epitélio superficial ovariano das mesmas doadoras. A atividade enzimática foi também avaliada em fibroblastos previamente induzidos a senescência com peróxido de hidrogeno. Resultados: as leituras da atividade enzimática, analisada em conjunto com a capacidade replicativa, indicam que os cultivos de fibroblastos alcançaram o estado senescente para os passes 4-5, o que também ocorreu com as células epiteliais. Os fibroblastos induzidos a senescência mostraram valores variáveis de atividade enzimática. Conclusões: as semelhanças entre os fibroblastos e as células epiteliais em quanto ao inicio da senescência poderia estar relacionado com a transição epitélio-mesenquimática que tem sido descrita como fator de risco de câncer derivado do epitélio superficial ovariano. Valores baixos de atividade β-galactosidase poderiam sugerir que, em alguns casos, ocorreu inativação das vias de resposta ao estresse oxidativo. 2011-05-27 2011-05-27 2014-07-09T15:55:58Z info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://revistas.urosario.edu.co/index.php/revsalud/article/view/1545 http://repository.urosario.edu.co/handle/10336/7396 spa http://creativecommons.org/licenses/by-nc/4.0 info:eu-repo/semantics/openAccess application/pdf Universidad del Rosario Revista Ciencias de la Salud; Vol. 9, núm. 1 (2011); 17-31 Revista Ciencias de la Salud; Vol. 9, núm. 1 (2011); 17-31 Revista Ciencias de la Salud; Vol. 9, núm. 1 (2011); 17-31 2145-4507 1692-7273 reponame:Repositorio Institucional EdocUR instname:Universidad del Rosario
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