Obtaining and characterizing extracellular vesicles from human prostate stromal cells derived from patients with benign prostatic hyperplasia
Benign Prostatic Hyperplasia (BPH) is the result of excessive cellullar proliferation in the prostatic transition zone, that compresses the urethra, causing the symptoms of the disease. Evidence supports that atherogenic environment could contribute to the development and progression of BPH, increas...
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| Autores principales: | , , , , |
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| Formato: | Artículo revista |
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Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
2022
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| Acceso en línea: | https://revistas.unc.edu.ar/index.php/med/article/view/38980 |
| Aporte de: |
| Sumario: | Benign Prostatic Hyperplasia (BPH) is the result of excessive cellullar proliferation in the prostatic transition zone, that compresses the urethra, causing the symptoms of the disease. Evidence supports that atherogenic environment could contribute to the development and progression of BPH, increasing its incidence and aggressiveness. The BPH hyperproliferative state requires coordinated communication between the different components that maintain a permissive microenvironment. Thus, extracellular vesicles (EVs) have become relevant as intercellular communication regulators in multiple processes. It has been described that oxidized-LDL molecule (OxLDL) would be involved in numerous signaling pathways; including signaling by EVs promoting cellullar proliferation. EVs, may contain diverse biomolecules that confer them functions in cellular communication. In addition, it has been reported that EVs derived from different cell types are capable to mediate proliferative and inflammatory effects.
In this context, we proposed as objectives: 1) to evaluate the effect of OxLDL, simulating an atherogenic state, on cell proliferation in primary cultures of human prostate stromal cells (HPSC) from patient samples (n=8) from the Sanatorio Allende of Córdoba; 2) to isolate EVs from primary cultures by differential ultracentrifugation and analyze their production and release into the medium in treatments with OxLDL vs. vehicle; 3) to morphologically characterize EVs by transmission electron microscopy (TEM) and confirm the identity and presence of exosomes, through CD63 immuno-staining using colloidal gold.
It was observed that OxLDL (20μM) produced a significant increase in cell proliferation rate compared to vehicle. EVs were obtained by differential ultracentrifugations (2k, 10k, 150k pellets) and visualized by TEM using negative staining. HPSC from patients with BPH showed a very low frequency of EVs released, with OxLDL inducing a 10-fold increase, especially in the 15-20nm fraction (p<0.001). Ultrastructurally, these EVs exhibited a spherical and concave appearance, compatible with exosomes. Therefore, they were positively verified by CD63 immunostaining.
Finally, we conclude that OxLDL would favor cell proliferation, increase and release of EVs, which would participate in cell communication and maintenance of the permissive environment, propitious to progression and increased aggressiveness of BPH. |
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