Homeostatic effects of testosterone in the hypoxic microenvironment of Benign Prostatic Hyperplasia

Benign prostatic hyperplasia (BPH) is characterized by an increase in stromal cell proliferation of the transitional zone of the prostate, with evidence suggesting that this is associated with a hypoxic microenvironment, which would induce proliferation through the transcription factor HIF-1. In pro...

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Autores principales: Cuello Rubio , MM, Peinetti , N, López Seoane, M, Maldonado, CA, Quintar , AA
Formato: Artículo revista
Lenguaje:Español
Publicado: Universidad Nacional Córdoba. 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/25655
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id I10-R327-article-25655
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
language Español
format Artículo revista
topic benign prostatic hyperplasia
testosterone
hypoxia
hiperplasia protática benigna
testosterona
hipoxia
spellingShingle benign prostatic hyperplasia
testosterone
hypoxia
hiperplasia protática benigna
testosterona
hipoxia
Cuello Rubio , MM
Peinetti , N
López Seoane, M
Maldonado, CA
Quintar , AA
Homeostatic effects of testosterone in the hypoxic microenvironment of Benign Prostatic Hyperplasia
topic_facet benign prostatic hyperplasia
testosterone
hypoxia
hiperplasia protática benigna
testosterona
hipoxia
author Cuello Rubio , MM
Peinetti , N
López Seoane, M
Maldonado, CA
Quintar , AA
author_facet Cuello Rubio , MM
Peinetti , N
López Seoane, M
Maldonado, CA
Quintar , AA
author_sort Cuello Rubio , MM
title Homeostatic effects of testosterone in the hypoxic microenvironment of Benign Prostatic Hyperplasia
title_short Homeostatic effects of testosterone in the hypoxic microenvironment of Benign Prostatic Hyperplasia
title_full Homeostatic effects of testosterone in the hypoxic microenvironment of Benign Prostatic Hyperplasia
title_fullStr Homeostatic effects of testosterone in the hypoxic microenvironment of Benign Prostatic Hyperplasia
title_full_unstemmed Homeostatic effects of testosterone in the hypoxic microenvironment of Benign Prostatic Hyperplasia
title_sort homeostatic effects of testosterone in the hypoxic microenvironment of benign prostatic hyperplasia
description Benign prostatic hyperplasia (BPH) is characterized by an increase in stromal cell proliferation of the transitional zone of the prostate, with evidence suggesting that this is associated with a hypoxic microenvironment, which would induce proliferation through the transcription factor HIF-1. In prostate cancer, HIF-1 is highly regulated by testosterone but little is known about the role of androgens on HIF-1 in BPH. Our objective was therefore to evaluate the effect of hypoxia on stromal cell proliferation and phenotype and the regulatory role of testosterone. Prostatic stromal cells harvested from patients with BPH (n=10, approved by the Bioethics Committee of Sanatorio Allende) were cultured and stimulated with CoCl2 (200 µM), a stabilizer of HIF-1 that mimics hypoxia, alone or in combination with testosterone (T) at high (10 µM), physiological (0.1 µM), and low (1nM) concentrations for 24 hours. The expression and nuclear translocation of HIF-1 were measured by western blot and immunofluorescence. Cell proliferation was assessed by immunocytochemistry of Ki67 and BrDu incorporation, whereas the cellular phenotype was confirmed by transmission electron microscopy (TEM) and western blot of cytoskeletal markers for stromal cells. CoCl2 upregulated the expression and nuclear translocation of HIF-1, associated with increased cell proliferation (3.10 ± 0.17% vs. CoCl2: 10.77 ± 4.51%, p <0.05). The presence of testosterone at low and physiological doses reduced these parameters, indicating a protective role of the androgen in hypoxic conditions (CoCl2 + T [0.1 µM]: 5.59 ± 2.25%). In control cells, TEM analysis showed signs of activation such as cellular edema, dilation of the endoplasmic reticulum, with multiple foci of contractile myofibrils, compatible with a myofibroblastic profile. Hypoxia increased these signs, with frequent myocontractile fibrils, while the presence of testosterone maintained a cell phenotype similar to the controls. Our results suggest that a hypoxic microenvironment promotes a HIF-1-mediated hyperproliferative cell phenotype in BPH, with testosterone modulating the cellular response to hypoxia. Therefore, HIF-1 inhibition would be likely a molecular mechanism involved in the homeostatic effects of testosterone in this pathology.
publisher Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
publishDate 2019
url https://revistas.unc.edu.ar/index.php/med/article/view/25655
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spelling I10-R327-article-256552024-08-27T18:26:01Z Homeostatic effects of testosterone in the hypoxic microenvironment of Benign Prostatic Hyperplasia Efectos homeostáticos de testosterona en el contexto hipóxico de la hiperplasia prostática benigna Cuello Rubio , MM Peinetti , N López Seoane, M Maldonado, CA Quintar , AA benign prostatic hyperplasia testosterone hypoxia hiperplasia protática benigna testosterona hipoxia Benign prostatic hyperplasia (BPH) is characterized by an increase in stromal cell proliferation of the transitional zone of the prostate, with evidence suggesting that this is associated with a hypoxic microenvironment, which would induce proliferation through the transcription factor HIF-1. In prostate cancer, HIF-1 is highly regulated by testosterone but little is known about the role of androgens on HIF-1 in BPH. Our objective was therefore to evaluate the effect of hypoxia on stromal cell proliferation and phenotype and the regulatory role of testosterone. Prostatic stromal cells harvested from patients with BPH (n=10, approved by the Bioethics Committee of Sanatorio Allende) were cultured and stimulated with CoCl2 (200 µM), a stabilizer of HIF-1 that mimics hypoxia, alone or in combination with testosterone (T) at high (10 µM), physiological (0.1 µM), and low (1nM) concentrations for 24 hours. The expression and nuclear translocation of HIF-1 were measured by western blot and immunofluorescence. Cell proliferation was assessed by immunocytochemistry of Ki67 and BrDu incorporation, whereas the cellular phenotype was confirmed by transmission electron microscopy (TEM) and western blot of cytoskeletal markers for stromal cells. CoCl2 upregulated the expression and nuclear translocation of HIF-1, associated with increased cell proliferation (3.10 ± 0.17% vs. CoCl2: 10.77 ± 4.51%, p <0.05). The presence of testosterone at low and physiological doses reduced these parameters, indicating a protective role of the androgen in hypoxic conditions (CoCl2 + T [0.1 µM]: 5.59 ± 2.25%). In control cells, TEM analysis showed signs of activation such as cellular edema, dilation of the endoplasmic reticulum, with multiple foci of contractile myofibrils, compatible with a myofibroblastic profile. Hypoxia increased these signs, with frequent myocontractile fibrils, while the presence of testosterone maintained a cell phenotype similar to the controls. Our results suggest that a hypoxic microenvironment promotes a HIF-1-mediated hyperproliferative cell phenotype in BPH, with testosterone modulating the cellular response to hypoxia. Therefore, HIF-1 inhibition would be likely a molecular mechanism involved in the homeostatic effects of testosterone in this pathology. La hiperplasia prostática benigna (BPH) es una patología caracterizada por una proliferación celular estromal excesiva en la zona de transición prostática. Evidencias sugieren que la BPH está asociada a un microambiente hipóxico, el cual induciría proliferación mediante el factor de transcripción HIF-1. En cáncer de próstata, testosterona modula a HIF-1 pero se desconoce el rol de esta proteína y su regulación en patologías de crecimiento benigno. Nuestro objetivo fue examinar in vitro el efecto de la hipoxia sobre la proliferación y el fenotipo de células estromales de pacientes con BPH y analizar el rol modulador de testosterona en esta condición. Se utilizaron cultivos primarios de células estromales prostáticas provenientes de muestras de pacientes con BPH (n = 10) (aprobado por el Comité de Bioética del Sanatorio Allende), se estimularon con CoCl2 (200 µM), un imitador de hipoxia que estabiliza a HIF-1. A su vez, las células fueron coestimuladas con testosterona a alta (10µM), fisiológica (0.1µM), y baja dosis (1nM) por 24hs. Se determinó la expresión de HIF-1 y su translocación nuclear por western blot e inmunoflorescencia. La proliferación celular fue analizada por KI67 Y BrdU, mientras que el fenotipo fue evaluado por microscopía electrónica (TEM), inmunofluorescencia y western blot de marcadores musculares y fibroblásticos. CoCl2 incrementó la expresión y la translocación nuclear de HIF-1, asociándose con una mayor proliferación celular (3.10±0,17 vs. CoCl2: 10,77±4,51%, p<0,05). Testosterona (T) redujo estos parámetros tanto en dosis baja como en fisiológica, indicando un rol protector de testosterona en condiciones de hipoxia (CoCl2+T [0,1 µM]: 5.59±2.25%). Las células estromales de pacientes con BPH mostraron signos de activación y proteinopoiesis por TEM, con múltiples focos de miofibrillas contráctiles compactibles con un fenotipo miofibroblástico. La hipoxia acentuó estos signos, mientras que testosterona a dosis fisiológicas retornó a las células a un fenotipo similar al control. Nuestros resultados sugieren que un microambiente hipóxico promueve un estado hiperproliferativo y un fenotipo celular reactivo en la BPH, mediados por la actividad de HIF-1. Testosterona modula la respuesta celular a hipoxia, siendo la inhibición de HIF-1 un mecanismo molecular probablemente involucrado en los efectos homeostáticos de testosterona en esta patología. Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2019-10-10 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf https://revistas.unc.edu.ar/index.php/med/article/view/25655 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 10.31053/1853.0605.v76.nSuplemento spa https://revistas.unc.edu.ar/index.php/med/article/view/25655/27372 Derechos de autor 2019 Universidad Nacional de Córdoba https://creativecommons.org/licenses/by-nc/4.0