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miR-424-5p regulates apoptosis and cell proliferation via targeting Bcl2 in nucleus pulposus cells

miRNAs play an important role in the pathogenesis of intervertebral disc degeneration (IDD). The role and the underlying mechanism of miR-424-5p in human nucleus pulposus (NP) are still unknown. We aimed to explore the role of miR-424-5p in IDD. Real-time PCR was used to detect the expression of miR...

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Autores principales: Lu, Hua-tuo, Xu, Yong-qing, Wang, Hai, Zhang, Xu-lin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7651851/
https://www.ncbi.nlm.nih.gov/pubmed/33209193
http://dx.doi.org/10.1080/19768354.2020.1775699
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author Lu, Hua-tuo
Xu, Yong-qing
Wang, Hai
Zhang, Xu-lin
author_facet Lu, Hua-tuo
Xu, Yong-qing
Wang, Hai
Zhang, Xu-lin
author_sort Lu, Hua-tuo
collection PubMed
description miRNAs play an important role in the pathogenesis of intervertebral disc degeneration (IDD). The role and the underlying mechanism of miR-424-5p in human nucleus pulposus (NP) are still unknown. We aimed to explore the role of miR-424-5p in IDD. Real-time PCR was used to detect the expression of miR-424-5p and Bcl2 in IDD tissues and idiopathic scoliosis tissues. Human NP cells were used in our study. MTT and Hoechst apoptosis assays were used to detect the proliferation and apoptosis of NP cells, respectively. Western blotting assays were used to detect the expression levels of Bcl-2, cleaved caspase-3, cleaved caspase-9, caspase-3 and caspase-9 in degenerative NP cells. A luciferase reporter assay was applied to confirm the relationship between miR-424-5p and Bcl2. Our results showed that the expression of miR-424-5p was increased and Bcl2 was decreased in degenerative NP cells. miR-425-5p expression was negatively correlated with Bcl2 expression in IDD tissues. Suppression of miR-424-5p using an inhibitor increased Bcl2 expression at both the mRNA and protein levels, and it promoted cell viability and inhibited apoptosis. Furthermore, the levels of cleaved caspase-3 and cleaved caspase-9 were downregulated in miR-424-5p-silenced NP cells. Interestingly, we found that silencing miR-424-5p increased p62 expression at both the mRNA and protein levels. Finally, a luciferase reporter assay verified the binding of the miR-424-5p and the 3’UTR of Bcl2. These results suggested that silencing miR-424-5p suppressed NP cell apoptosis by upregulating Bcl2. Therefore, miR-424-5p might be a novel target for IDD therapies.
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spelling pubmed-76518512020-11-17 miR-424-5p regulates apoptosis and cell proliferation via targeting Bcl2 in nucleus pulposus cells Lu, Hua-tuo Xu, Yong-qing Wang, Hai Zhang, Xu-lin Anim Cells Syst (Seoul) Molecular Cellular Biology miRNAs play an important role in the pathogenesis of intervertebral disc degeneration (IDD). The role and the underlying mechanism of miR-424-5p in human nucleus pulposus (NP) are still unknown. We aimed to explore the role of miR-424-5p in IDD. Real-time PCR was used to detect the expression of miR-424-5p and Bcl2 in IDD tissues and idiopathic scoliosis tissues. Human NP cells were used in our study. MTT and Hoechst apoptosis assays were used to detect the proliferation and apoptosis of NP cells, respectively. Western blotting assays were used to detect the expression levels of Bcl-2, cleaved caspase-3, cleaved caspase-9, caspase-3 and caspase-9 in degenerative NP cells. A luciferase reporter assay was applied to confirm the relationship between miR-424-5p and Bcl2. Our results showed that the expression of miR-424-5p was increased and Bcl2 was decreased in degenerative NP cells. miR-425-5p expression was negatively correlated with Bcl2 expression in IDD tissues. Suppression of miR-424-5p using an inhibitor increased Bcl2 expression at both the mRNA and protein levels, and it promoted cell viability and inhibited apoptosis. Furthermore, the levels of cleaved caspase-3 and cleaved caspase-9 were downregulated in miR-424-5p-silenced NP cells. Interestingly, we found that silencing miR-424-5p increased p62 expression at both the mRNA and protein levels. Finally, a luciferase reporter assay verified the binding of the miR-424-5p and the 3’UTR of Bcl2. These results suggested that silencing miR-424-5p suppressed NP cell apoptosis by upregulating Bcl2. Therefore, miR-424-5p might be a novel target for IDD therapies. Taylor & Francis 2020-06-23 /pmc/articles/PMC7651851/ /pubmed/33209193 http://dx.doi.org/10.1080/19768354.2020.1775699 Text en © 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Molecular Cellular Biology
Lu, Hua-tuo
Xu, Yong-qing
Wang, Hai
Zhang, Xu-lin
miR-424-5p regulates apoptosis and cell proliferation via targeting Bcl2 in nucleus pulposus cells
title miR-424-5p regulates apoptosis and cell proliferation via targeting Bcl2 in nucleus pulposus cells
title_full miR-424-5p regulates apoptosis and cell proliferation via targeting Bcl2 in nucleus pulposus cells
title_fullStr miR-424-5p regulates apoptosis and cell proliferation via targeting Bcl2 in nucleus pulposus cells
title_full_unstemmed miR-424-5p regulates apoptosis and cell proliferation via targeting Bcl2 in nucleus pulposus cells
title_short miR-424-5p regulates apoptosis and cell proliferation via targeting Bcl2 in nucleus pulposus cells
title_sort mir-424-5p regulates apoptosis and cell proliferation via targeting bcl2 in nucleus pulposus cells
topic Molecular Cellular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7651851/
https://www.ncbi.nlm.nih.gov/pubmed/33209193
http://dx.doi.org/10.1080/19768354.2020.1775699
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