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Induction of osteoclast formation by LOX mutant (LOXG473A) through regulation of autophagy

BACKGROUND: Lysyl oxidase (LOX) has been identified to modulate osteoclast activity, so we explored the role of LOX(G473A), the highest frequency single nucleotide polymorphism in LOX, in osteoclast formation and its potential relationship to autophagy. METHODS: The ability of the LOX mutant, LOX(G4...

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Autores principales: Zhang, Bo, Luo, Chenglin, Xiao, Wenjin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AME Publishing Company 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8506719/
https://www.ncbi.nlm.nih.gov/pubmed/34734026
http://dx.doi.org/10.21037/atm-21-4474
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author Zhang, Bo
Luo, Chenglin
Xiao, Wenjin
author_facet Zhang, Bo
Luo, Chenglin
Xiao, Wenjin
author_sort Zhang, Bo
collection PubMed
description BACKGROUND: Lysyl oxidase (LOX) has been identified to modulate osteoclast activity, so we explored the role of LOX(G473A), the highest frequency single nucleotide polymorphism in LOX, in osteoclast formation and its potential relationship to autophagy. METHODS: The ability of the LOX mutant, LOX(G473A), to promote autophagy and osteoclast formation was evaluated in the pre-osteoclast cell line RAW264.7. Furthermore, autophagy-related protein expression and autophagosomes were detected by western blot and electron microscopy, respectively. Simultaneously, osteoclast formation and resorption ability were also detected using TRAP staining assay and bone resorption assay. In addition, the osteoclast-related proteins and mRNAs, as well as p-AMPKα and p-mTOR proteins, were further evaluated by western blot and qPCR assays. RESULTS: Autophagy inhibitor 3-MA suppressed the Beclin-1 and ATG5 protein levels and the ratio of LC3-II to LC3-I, as well as autophagosome formation in RAW264.7 transfected with the MUT plasmid and enhanced p62 protein expression. Simultaneously, 3-MA also reduced osteoclast formation and resorption, as well as the F-actin ring level of osteoclasts. In addition, 3-MA inhibited osteoclast-related protein and mRNA expression, including NFATC1, ACP5, CTSK. And the autophagy-related pathway protein p-AMPKα was increased and p-mTOR was reduced by 3-MA treatment. However, autophagy agonist RAPA reversed the effect of 3-MA on RAW264.7 with LOX(G473A) mutation, indicating that promoting autophagy could enhance the ability of LOX(G473A) to induce osteoclast formation. CONCLUSIONS: LOX mutant (LOX(G473A)) might promote osteoclast formation for RAW264.7 by enhancing autophagy via the AMPK/mTOR pathway, which is a new direction for bone disease research.
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spelling pubmed-85067192021-11-02 Induction of osteoclast formation by LOX mutant (LOXG473A) through regulation of autophagy Zhang, Bo Luo, Chenglin Xiao, Wenjin Ann Transl Med Original Article BACKGROUND: Lysyl oxidase (LOX) has been identified to modulate osteoclast activity, so we explored the role of LOX(G473A), the highest frequency single nucleotide polymorphism in LOX, in osteoclast formation and its potential relationship to autophagy. METHODS: The ability of the LOX mutant, LOX(G473A), to promote autophagy and osteoclast formation was evaluated in the pre-osteoclast cell line RAW264.7. Furthermore, autophagy-related protein expression and autophagosomes were detected by western blot and electron microscopy, respectively. Simultaneously, osteoclast formation and resorption ability were also detected using TRAP staining assay and bone resorption assay. In addition, the osteoclast-related proteins and mRNAs, as well as p-AMPKα and p-mTOR proteins, were further evaluated by western blot and qPCR assays. RESULTS: Autophagy inhibitor 3-MA suppressed the Beclin-1 and ATG5 protein levels and the ratio of LC3-II to LC3-I, as well as autophagosome formation in RAW264.7 transfected with the MUT plasmid and enhanced p62 protein expression. Simultaneously, 3-MA also reduced osteoclast formation and resorption, as well as the F-actin ring level of osteoclasts. In addition, 3-MA inhibited osteoclast-related protein and mRNA expression, including NFATC1, ACP5, CTSK. And the autophagy-related pathway protein p-AMPKα was increased and p-mTOR was reduced by 3-MA treatment. However, autophagy agonist RAPA reversed the effect of 3-MA on RAW264.7 with LOX(G473A) mutation, indicating that promoting autophagy could enhance the ability of LOX(G473A) to induce osteoclast formation. CONCLUSIONS: LOX mutant (LOX(G473A)) might promote osteoclast formation for RAW264.7 by enhancing autophagy via the AMPK/mTOR pathway, which is a new direction for bone disease research. AME Publishing Company 2021-09 /pmc/articles/PMC8506719/ /pubmed/34734026 http://dx.doi.org/10.21037/atm-21-4474 Text en 2021 Annals of Translational Medicine. All rights reserved. https://creativecommons.org/licenses/by-nc-nd/4.0/Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0 (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Original Article
Zhang, Bo
Luo, Chenglin
Xiao, Wenjin
Induction of osteoclast formation by LOX mutant (LOXG473A) through regulation of autophagy
title Induction of osteoclast formation by LOX mutant (LOXG473A) through regulation of autophagy
title_full Induction of osteoclast formation by LOX mutant (LOXG473A) through regulation of autophagy
title_fullStr Induction of osteoclast formation by LOX mutant (LOXG473A) through regulation of autophagy
title_full_unstemmed Induction of osteoclast formation by LOX mutant (LOXG473A) through regulation of autophagy
title_short Induction of osteoclast formation by LOX mutant (LOXG473A) through regulation of autophagy
title_sort induction of osteoclast formation by lox mutant (loxg473a) through regulation of autophagy
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8506719/
https://www.ncbi.nlm.nih.gov/pubmed/34734026
http://dx.doi.org/10.21037/atm-21-4474
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AT xiaowenjin inductionofosteoclastformationbyloxmutantloxg473athroughregulationofautophagy