Pro-oxidant effects induced by 17β-estradiol promote p53 and p21 activation in normal and tumoral pituitary cells

Abstract:  The estrogenic action on pituitary cell growth is widely known. We have previously demonstrated the activation of the antioxidant pathway of phosphorylated erythroid 2-derived nuclear factor 2 (p-Nrf2) in response to DNA damage by 17β-estradiol (E2). In this study we analyze...

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Autores principales: Grondona , E, Venier , AC, Sosa , LDV, Pesaola , F, Fernandez , S, Sabatino , ME, Gutiérrez , S, De Paul, AL
Formato: Artículo revista
Publicado: Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2021
Materias:
P53
P21
p53
p21
.
Acceso en línea:https://revistas.unc.edu.ar/index.php/med/article/view/35080
Aporte de:
id I10-R327-article-35080
record_format ojs
institution Universidad Nacional de Córdoba
institution_str I-10
repository_str R-327
container_title_str Revista de la Facultad de Ciencias Médicas de Córdoba
format Artículo revista
topic Estrogen
Pituitary
P53
P21
oxidative stress
estrógeno
Hipófisis
p53
p21
Estrés Oxidativo
.
spellingShingle Estrogen
Pituitary
P53
P21
oxidative stress
estrógeno
Hipófisis
p53
p21
Estrés Oxidativo
.
Grondona , E
Venier , AC
Sosa , LDV
Pesaola , F
Fernandez , S
Sabatino , ME
Gutiérrez , S
De Paul, AL
Pro-oxidant effects induced by 17β-estradiol promote p53 and p21 activation in normal and tumoral pituitary cells
topic_facet Estrogen
Pituitary
P53
P21
oxidative stress
estrógeno
Hipófisis
p53
p21
Estrés Oxidativo
.
author Grondona , E
Venier , AC
Sosa , LDV
Pesaola , F
Fernandez , S
Sabatino , ME
Gutiérrez , S
De Paul, AL
author_facet Grondona , E
Venier , AC
Sosa , LDV
Pesaola , F
Fernandez , S
Sabatino , ME
Gutiérrez , S
De Paul, AL
author_sort Grondona , E
title Pro-oxidant effects induced by 17β-estradiol promote p53 and p21 activation in normal and tumoral pituitary cells
title_short Pro-oxidant effects induced by 17β-estradiol promote p53 and p21 activation in normal and tumoral pituitary cells
title_full Pro-oxidant effects induced by 17β-estradiol promote p53 and p21 activation in normal and tumoral pituitary cells
title_fullStr Pro-oxidant effects induced by 17β-estradiol promote p53 and p21 activation in normal and tumoral pituitary cells
title_full_unstemmed Pro-oxidant effects induced by 17β-estradiol promote p53 and p21 activation in normal and tumoral pituitary cells
title_sort pro-oxidant effects induced by 17β-estradiol promote p53 and p21 activation in normal and tumoral pituitary cells
description Abstract:  The estrogenic action on pituitary cell growth is widely known. We have previously demonstrated the activation of the antioxidant pathway of phosphorylated erythroid 2-derived nuclear factor 2 (p-Nrf2) in response to DNA damage by 17β-estradiol (E2). In this study we analyzed the impact of E2 on the tumoral suppressor p53 and cell cycle regulator p21 activation, as well as the damage response in pituitary cells in vitro. Pituitary tumour development was induced in adult male Wistar rats by subcutaneous implantation of silastic capsules containing estradiol benzoate (30mg) for 10 days (E10; n = 5). The control group was implanted with empty capsules (n=5). Subsequently, pituitary glands were collected, with cells being cultured and exposed to E2 (1-10-100nM) for 15, 30 and 60 min. The p53 and p21 protein levels were determined by western blot. By immunofluorescence, the co-expression of prolactin (PRL)/p-Nrf2 and growth hormone (GH)/p-Nrf2 was evaluated to determinate the cell type involved in the activation of the oxidative damage response. Statistical analysis: ANOVA-Fischer (p <0.05). Under tumoral contexts, a significant increase in p53 protein at the cytoplasmic level was detected after 15 and 30 min of treatment with E2 (1nM). After supraphysiological concentrations (10-100nM), this response was observed after 30 and 60 min. At nuclear level, we only detected an increase in p53 at 15 min, regardless of the dose. The p2 expression showed a similar profile in both subcellular compartments, with significant increases after 15 and 30 min of exposure with 1nM E2 and 30 and 60 min with supraphysiological doses. In normal cells cultures we did not observe significant changes in the expression of both markers. The number of lactotroph tumoral cells co-expressing PRL/p-Nrf2 increased significantly at 30 min compared to supraphysiological doses of E2. No changes were detected in the expression of GH/p-Nrf2 in GH cells. In tumoral pituitary cells, the pro-oxidant action induced by E2 triggers the activation of p53 and p21 in order to repair DNA damage, through the stabilization of Nrf2. This response would mainly have an impact on PRL tumoral cells. These mechanisms could guarantee the cell viability, thus regulating pituitary tumor development.
publisher Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
publishDate 2021
url https://revistas.unc.edu.ar/index.php/med/article/view/35080
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first_indexed 2024-09-03T21:03:09Z
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spelling I10-R327-article-350802024-04-15T16:19:09Z Pro-oxidant effects induced by 17β-estradiol promote p53 and p21 activation in normal and tumoral pituitary cells Efectos pro-oxidantes inducidos por 17β-estradiol promueven la activación de p53 y p21 en células hipofisarias normales y tumorales A Grondona , E Venier , AC Sosa , LDV Pesaola , F Fernandez , S Sabatino , ME Gutiérrez , S De Paul, AL Estrogen Pituitary P53 P21 oxidative stress estrógeno Hipófisis p53 p21 Estrés Oxidativo . Abstract:  The estrogenic action on pituitary cell growth is widely known. We have previously demonstrated the activation of the antioxidant pathway of phosphorylated erythroid 2-derived nuclear factor 2 (p-Nrf2) in response to DNA damage by 17β-estradiol (E2). In this study we analyzed the impact of E2 on the tumoral suppressor p53 and cell cycle regulator p21 activation, as well as the damage response in pituitary cells in vitro. Pituitary tumour development was induced in adult male Wistar rats by subcutaneous implantation of silastic capsules containing estradiol benzoate (30mg) for 10 days (E10; n = 5). The control group was implanted with empty capsules (n=5). Subsequently, pituitary glands were collected, with cells being cultured and exposed to E2 (1-10-100nM) for 15, 30 and 60 min. The p53 and p21 protein levels were determined by western blot. By immunofluorescence, the co-expression of prolactin (PRL)/p-Nrf2 and growth hormone (GH)/p-Nrf2 was evaluated to determinate the cell type involved in the activation of the oxidative damage response. Statistical analysis: ANOVA-Fischer (p <0.05). Under tumoral contexts, a significant increase in p53 protein at the cytoplasmic level was detected after 15 and 30 min of treatment with E2 (1nM). After supraphysiological concentrations (10-100nM), this response was observed after 30 and 60 min. At nuclear level, we only detected an increase in p53 at 15 min, regardless of the dose. The p2 expression showed a similar profile in both subcellular compartments, with significant increases after 15 and 30 min of exposure with 1nM E2 and 30 and 60 min with supraphysiological doses. In normal cells cultures we did not observe significant changes in the expression of both markers. The number of lactotroph tumoral cells co-expressing PRL/p-Nrf2 increased significantly at 30 min compared to supraphysiological doses of E2. No changes were detected in the expression of GH/p-Nrf2 in GH cells. In tumoral pituitary cells, the pro-oxidant action induced by E2 triggers the activation of p53 and p21 in order to repair DNA damage, through the stabilization of Nrf2. This response would mainly have an impact on PRL tumoral cells. These mechanisms could guarantee the cell viability, thus regulating pituitary tumor development. Resumen:  Es ampliamente conocida la acción estrogénica sobre el crecimiento celular hipofisario. Previamente demostramos la activación de la vía antioxidante del factor nuclear 2 derivado del eritroide 2 fosforilado (Nrf2-p) en respuesta al daño del ADN por 17β-estradiol (E2). Analizamos entonces, el impacto del E2 sobre la activación del supresor tumoral p53 y el regulador del ciclo celular p21, y la respuesta al daño en células hipofisarias in vitro. Indujimos el desarrollo tumoral hipofisario en ratas Wistar macho adultas mediante la implantación subcutánea de cápsulas de silástico conteniendo benzoato de estradiol (30 mg) durante 10 días (E10; n=5). El grupo control fue implantado con cápsulas vacía (n=5). Posteriormente, extrajimos las adenohipófisis, cultivamos sus células y fueron expuestas a E2 (1-10-100nM) por 15, 30 y 60min. Determinamos los niveles proteicos de p53 y p21 por western blot. Por inmunofluorescencia, evaluamos la co-expresión de Prolactina (PRL)/Nrf2-p y hormona de crecimiento (GH)/Nrf2-p para determinar el tipo celular involucrado con activación de respuesta al daño oxidativo. Análisis estadístico: ANOVA-Fischer (p <0,05). En contextos tumorales, detectamos un aumento significativo de la proteína p53 a nivel citoplasmático luego de 15 y 30 min de tratamiento con E2 (1nM); frente a concentraciones suprafisiológicas (10-100nM) esta respuesta se observó luego de 30 y 60 min. A nivel nuclear, detectamos un aumento de p53 solo a los 15 min, de manera independiente de la dosis. La expresión de p21 mostró un perfil similar en ambos compartimientos subcelulares con incrementos significativos luego de 15 y 30 min de exposición con E2 1nM y 30 y 60 min con dosis suprafisiológicas. En cultivos normales no observamos cambios significativos en la expresión de ambos marcadores. El número de células lactotropas tumorales que co-expresaron PRL/Nrf2-p aumentó significativamente a los 30 min frente a dosis suprafisiológicas de E2; sin detectarse modificaciones en la expresión de GH/Nrf2-p en células somatotropas. En células hipofisarias tumorales, la acción pro-oxidante inducida por E2 desencadena la activación de p53 y de p21 a fin de reparar el daño del ADN, vía estabilización de Nrf2. Esta respuesta impactaría principalmente sobre células lactotropas tumorales. Mediante estos mecanismos se garantizaría la viabilidad celular, regulando el desarrollo tumoral hipofisario. . Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2021-10-12 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion texto texto texto https://revistas.unc.edu.ar/index.php/med/article/view/35080 Revista de la Facultad de Ciencias Médicas de Córdoba.; Vol. 78 No. Suplemento (2021): Suplemento JIC XXII Revista de la Facultad de Ciencias Médicas de Córdoba; Vol. 78 Núm. Suplemento (2021): Suplemento JIC XXII Revista da Faculdade de Ciências Médicas de Córdoba; v. 78 n. Suplemento (2021): Suplemento JIC XXII 1853-0605 0014-6722 http://creativecommons.org/licenses/by-nc/4.0