Estrogen effects on the mitochondrial dynamic and oxidative damage response in normal and tumoral pituitary cells
We recently described the relationship between mitochondrial alterations and senescence as a regulatory process of cell growth in estrogen-induced pituitary tumors. In addition, it is known that this hormone promotes the DNA oxidation leading to the accumulation of mutations in mitochondrial DNA res...
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| Formato: | Artículo revista |
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Universidad Nacional Cba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
2019
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| Acceso en línea: | https://revistas.unc.edu.ar/index.php/med/article/view/25595 |
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I10-R10-article-25595 |
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ojs |
| institution |
Universidad Nacional de Córdoba |
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I-10 |
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R-10 |
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Revistas de la UNC |
| language |
Español |
| format |
Artículo revista |
| topic |
pituitary estrogen mitochondria oxidative stress hipófisis estrógeno mitocondria estrés oxidativo |
| spellingShingle |
pituitary estrogen mitochondria oxidative stress hipófisis estrógeno mitocondria estrés oxidativo Grondona , E Mongi Bragato, E Sosa, LDV Latini, A De Paul, AL Estrogen effects on the mitochondrial dynamic and oxidative damage response in normal and tumoral pituitary cells |
| topic_facet |
pituitary estrogen mitochondria oxidative stress hipófisis estrógeno mitocondria estrés oxidativo |
| author |
Grondona , E Mongi Bragato, E Sosa, LDV Latini, A De Paul, AL |
| author_facet |
Grondona , E Mongi Bragato, E Sosa, LDV Latini, A De Paul, AL |
| author_sort |
Grondona , E |
| title |
Estrogen effects on the mitochondrial dynamic and oxidative damage response in normal and tumoral pituitary cells |
| title_short |
Estrogen effects on the mitochondrial dynamic and oxidative damage response in normal and tumoral pituitary cells |
| title_full |
Estrogen effects on the mitochondrial dynamic and oxidative damage response in normal and tumoral pituitary cells |
| title_fullStr |
Estrogen effects on the mitochondrial dynamic and oxidative damage response in normal and tumoral pituitary cells |
| title_full_unstemmed |
Estrogen effects on the mitochondrial dynamic and oxidative damage response in normal and tumoral pituitary cells |
| title_sort |
estrogen effects on the mitochondrial dynamic and oxidative damage response in normal and tumoral pituitary cells |
| description |
We recently described the relationship between mitochondrial alterations and senescence as a regulatory process of cell growth in estrogen-induced pituitary tumors. In addition, it is known that this hormone promotes the DNA oxidation leading to the accumulation of mutations in mitochondrial DNA resulting in energy deficiencies and cellular damage with carcinogenic potential. In this context, we set out to evaluate possible 17β-estradiol (E2) pro-oxidant actions in vitro on mitochondrial dynamics and the activation of the damage response under pituitary tumor contexts.
Pituitary tumor development was induced in adult male Wistar rats by subcutaneous implantation of silastic capsules containing estradiol benzoate (30 mg) for 10 days (E10; n=5). The control group was implanted with empty capsules (n=5). Once the deadlines were met, pituitary glands were collected and cells cultured and exposed to E2 (1-10-100nM) for 15, 30 and 60 min. The reactive oxygen species (ROS) generation was analyzed by flow cytometry, the 8OHdG and p-Nrf2 co-expression, as indicators of DNA oxidation and activation of response to oxidative damage, was evaluated by immunofluorescence. Mitochondrial protein expression: MFN2 and OPA-1 (mitochondrial fusion) and p-Drp1 (mitochondrial fission) was assessed by western blot. Statistical analysis: ANOVA-Fischer (p <0.05).
In tumoral cells, physiological doses of E2 (1nM) promoted a significant increase in the ROS production after short exposure times, whereas in normal cells, 10nM E2 induced this oxidative response after 30min of treatment. Likewise, an increase in the co-expression of 8OHdG and p-Nrf2 was observed as the concentration and the time of exposure to E2 increased. This same behavior was observed in the expression of mitochondrial proteins, indicating a tendency towards fusion.
In tumoral cells, E2 would promote a pro-oxidant environment that would be counterbalanced by the activation of antioxidant pathways mediated by Nrf2. These clear evidences of oxidative stress would trigger alterations in mitochondrial dynamics, favoring the fusion process, as a measure of metabolic readaptation. These responses would guarantee cell viability allowing to meet the highest energy demands, typical of the tumor context. |
| publisher |
Universidad Nacional Cba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología |
| publishDate |
2019 |
| url |
https://revistas.unc.edu.ar/index.php/med/article/view/25595 |
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2022-08-20T01:26:33Z |
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2022-08-20T01:26:33Z |
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I10-R10-article-255952019-11-11T21:18:27Z Estrogen effects on the mitochondrial dynamic and oxidative damage response in normal and tumoral pituitary cells Efectos del estrógeno sobre la dinámica mitocondrial y la respuesta al daño oxidativo en células hipofisarias normales y tumorales Grondona , E Mongi Bragato, E Sosa, LDV Latini, A De Paul, AL pituitary estrogen mitochondria oxidative stress hipófisis estrógeno mitocondria estrés oxidativo We recently described the relationship between mitochondrial alterations and senescence as a regulatory process of cell growth in estrogen-induced pituitary tumors. In addition, it is known that this hormone promotes the DNA oxidation leading to the accumulation of mutations in mitochondrial DNA resulting in energy deficiencies and cellular damage with carcinogenic potential. In this context, we set out to evaluate possible 17β-estradiol (E2) pro-oxidant actions in vitro on mitochondrial dynamics and the activation of the damage response under pituitary tumor contexts. Pituitary tumor development was induced in adult male Wistar rats by subcutaneous implantation of silastic capsules containing estradiol benzoate (30 mg) for 10 days (E10; n=5). The control group was implanted with empty capsules (n=5). Once the deadlines were met, pituitary glands were collected and cells cultured and exposed to E2 (1-10-100nM) for 15, 30 and 60 min. The reactive oxygen species (ROS) generation was analyzed by flow cytometry, the 8OHdG and p-Nrf2 co-expression, as indicators of DNA oxidation and activation of response to oxidative damage, was evaluated by immunofluorescence. Mitochondrial protein expression: MFN2 and OPA-1 (mitochondrial fusion) and p-Drp1 (mitochondrial fission) was assessed by western blot. Statistical analysis: ANOVA-Fischer (p <0.05). In tumoral cells, physiological doses of E2 (1nM) promoted a significant increase in the ROS production after short exposure times, whereas in normal cells, 10nM E2 induced this oxidative response after 30min of treatment. Likewise, an increase in the co-expression of 8OHdG and p-Nrf2 was observed as the concentration and the time of exposure to E2 increased. This same behavior was observed in the expression of mitochondrial proteins, indicating a tendency towards fusion. In tumoral cells, E2 would promote a pro-oxidant environment that would be counterbalanced by the activation of antioxidant pathways mediated by Nrf2. These clear evidences of oxidative stress would trigger alterations in mitochondrial dynamics, favoring the fusion process, as a measure of metabolic readaptation. These responses would guarantee cell viability allowing to meet the highest energy demands, typical of the tumor context. Recientemente describimos la relación entre las alteraciones mitocondriales y la senescencia como proceso regulador de crecimiento celular en tumores hipofisarios inducidos por estrógeno. Además, se sabe que esta hormona promueve la oxidación del ADN conduciendo a la acumulación de mutaciones en el ADN mitocondrial traduciéndose en deficiencias de energía y daño celular con potencialidad carcinogénica. En este contexto, nos propusimos evaluar la posible acción pro-oxidante del 17β-estradiol (E2) in vitro sobre la dinámica mitocondrial y la activación de la respuesta al daño en contextos tumorales hipofisarios. Se indujo 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). Cumplido los plazos, las hipófisis fueron extraídas y sus células cultivadas y expuestas a E2 (1-10-100nM) por 15, 30 y 60 min. Se analizó la generación de especies reactivas de oxígeno (ROS) por citometría de flujo, la co-expresión de 8OHdG y p-Nrf2 como indicadores de oxidación del ADN y activación de respuesta al daño oxidativo por inmunofluorescencia. La expresión de proteínas mitocondriales: MFN2 y OPA-1 (fusión mitocondrial) y p-Drp1 (fisión mitocondrial) se evaluó por western blot. Análisis estadístico: ANOVA-Fischer (p <0,05). En células tumorales, dosis fisiológicas de E2 (1nM) promovieron un aumento significativo en la generación de ROS a partir de tiempos cortos de exposición, en tanto que, en células normales, E2 10nM indujo esta oxidante respuesta luego de 30min de tratamiento. Igualmente, se observó un aumento en la co-expresión de 8OHdG y p-Nrf2 acorde aumentó la concentración y el tiempo de exposición a E2. Este mismo comportamiento se observó en la expresión de las proteínas mitocondriales analizadas, señalando una tendencia hacia la fusión. En células tumorales, el E2 promovería un entorno pro-oxidante que sería contrabalanceado por la activación de vías antioxidantes mediada por Nrf2. Estas claras evidencias de estrés oxidativo desencadenarían alteraciones en la dinámica mitocondrial, favoreciendo el proceso de fusión como medida de readaptación metabólica. Estas respuestas garantizarían la viabilidad celular permitiendo hacer frente a las mayores demandas energéticas, propio del contextos tumoral. Universidad Nacional Cba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2019-10-04 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf https://revistas.unc.edu.ar/index.php/med/article/view/25595 Revista de la Facultad de Ciencias Médicas de Córdoba.; 2019: Suplemento JIC XX Revista de la Facultad de Ciencias Médicas de Córdoba; 2019: Suplemento JIC XX Revista da Faculdade de Ciências Médicas de Córdoba; 2019: Suplemento JIC XX 1853-0605 0014-6722 spa https://revistas.unc.edu.ar/index.php/med/article/view/25595/27355 Derechos de autor 2019 Universidad Nacional de Córdoba |