Rol de la dinámica mitocondrial en el proceso de migración en cáncer de próstata.
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Date
2022
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Publisher
Universidad de Concepción
Abstract
El cáncer de próstata es el segundo cáncer más diagnosticado en hombres y es de alto riesgo en etapas tardías, donde las células son capaces de realizar invasión y metástasis a través de un proceso conocido como transición epitelio mesenquimal (EMT). La dinámica mitocondrial, que incluye los procesos de fisión mediada por la proteína DRP1 y de fusión mediada por la proteína OPA1, se encuentra desbalanceada en distintos tipos de cáncer, con un predominio de la fisión mitocondrial que se relacionaría con mayor migración e invasión celular. Sin embargo, el rol de estas proteínas en el proceso de migración en células de cáncer de próstata aún no está claro. El objetivo de esta tesis es determinar el rol de DRP1 y OPA1 en el proceso de EMT y la migración en células de cáncer de próstata, procesos que son fundamentales para el desarrollo de metástasis. Se utilizó como modelo de estudio la línea celular C4-2B y como estrategia principal el uso de ARN de interferencia (siRNA) para el silenciamiento de DRP1 y OPA1. Inicialmente, se analizó el efecto del silenciamiento de estas proteínas en la morfología mitocondrial y la expresión de marcadores de EMT. De manera complementaria, lo anterior fue analizado utilizando también el inhibidor específico de DRP1, Mdivi-1. Posteriormente, se analizó el efecto del silenciamiento de DRP1 y OPA1 en la migración celular, por medio de ensayos de la herida, transwell y microscopía. Los resultados mostraron que el silenciamiento de DRP1 induce la fusión mitocondrial e inhibe el proceso de EMT y la migración celular, mientras que el silenciamiento de OPA1 promueve la fisión mitocondrial, la EMT y la migración celular. Además, el tratamiento con Mdivi-1 produce efectos similares a los obtenidos al silenciar DRP1. En conclusión, cambios en la dinámica mitocondrial mediados por DRP1 y OPA1 regulan la migración celular, la que es promovida por la fisión mitocondrial e inhibida por la fusión. Nuestros resultados permiten establecer las bases para proponer en el futuro, a DRP1 y OPA1, como posibles blancos terapéuticos en cáncer de próstata.
Prostate cancer is the second most common cancer in men worldwide and presents high risk in late stages, where cells are capable of invasion and metastasis through a process known as epithelial mesenchymal transition (EMT). Mitochondrial dynamics, involving the fission mediated by DRP1 and the fusion mediated by OPA1, has been described as unbalanced in different types of cancer, with a predominance of mitochondrial fission that would be related to higher cell migration and invasion. However, the role of these proteins in prostate cancer remains unclear. The aim of this thesis is to determinate the role of DRP1 and OPA1 in the EMT process and migration in prostate cancer cells, processes that are essential for metastasis development. The C4-2B cell line was used as model of study and the main strategy was the use of interfering RNA (siRNA) for the silencing of DRP1 and OPA1. Initially, the effect of silencing these proteins in mitochondrial morphology and EMT markers expression was analyzed. Complementary, the above was also analyzed using the specific inhibitor of DRP1, Mdivi-1. Then, the effect of silencing DRP1 and OPA1 in cell migration was analyzed by wound healing assay, transwell assay and microscopy. The results showed that DRP1 silencing induces mitochondrial fusion and inhibits EMT process and cell migration, while OPA1 silencing induces mitochondrial fission, EMT and cell migration. Furthermore, Mdivi-1 treatment produces similar effects to those obtained by silencing DRP1. In conclusion, changes in mitochondrial dynamics mediated by DRP1 and OPA1 regulate cell migration, which is induced by mitochondrial fission and inhibited by fusion. These results allow us to establish the bases to propose DRP1 and OPA1 as possible therapeutic targets in prostate cancer in the future.
Prostate cancer is the second most common cancer in men worldwide and presents high risk in late stages, where cells are capable of invasion and metastasis through a process known as epithelial mesenchymal transition (EMT). Mitochondrial dynamics, involving the fission mediated by DRP1 and the fusion mediated by OPA1, has been described as unbalanced in different types of cancer, with a predominance of mitochondrial fission that would be related to higher cell migration and invasion. However, the role of these proteins in prostate cancer remains unclear. The aim of this thesis is to determinate the role of DRP1 and OPA1 in the EMT process and migration in prostate cancer cells, processes that are essential for metastasis development. The C4-2B cell line was used as model of study and the main strategy was the use of interfering RNA (siRNA) for the silencing of DRP1 and OPA1. Initially, the effect of silencing these proteins in mitochondrial morphology and EMT markers expression was analyzed. Complementary, the above was also analyzed using the specific inhibitor of DRP1, Mdivi-1. Then, the effect of silencing DRP1 and OPA1 in cell migration was analyzed by wound healing assay, transwell assay and microscopy. The results showed that DRP1 silencing induces mitochondrial fusion and inhibits EMT process and cell migration, while OPA1 silencing induces mitochondrial fission, EMT and cell migration. Furthermore, Mdivi-1 treatment produces similar effects to those obtained by silencing DRP1. In conclusion, changes in mitochondrial dynamics mediated by DRP1 and OPA1 regulate cell migration, which is induced by mitochondrial fission and inhibited by fusion. These results allow us to establish the bases to propose DRP1 and OPA1 as possible therapeutic targets in prostate cancer in the future.
Description
Tesis presentada para optar al título de Bioquímico/a.
Keywords
Cáncer, Próstata, Mitocondrias, Metástasis, Anticuerpos