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Proteomic analysis of human periodontal ligament cells under hypoxia

BACKGROUND: The periodontal ligament is essential for homeostasis of periodontal tissue. A hypoxic milieu of the periodontal tissue is generated under periodontitis or during orthodontic treatment, which affects the periodontal and bone remodelling process. Here, we provide a comprehensive proteomic...

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Autores principales: Li, Qiwen, Luo, Tao, Lu, Wenxin, Yi, Xiaoxiao, Zhao, Zhihe, Liu, Jun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6717648/
https://www.ncbi.nlm.nih.gov/pubmed/31496921
http://dx.doi.org/10.1186/s12953-019-0151-2
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author Li, Qiwen
Luo, Tao
Lu, Wenxin
Yi, Xiaoxiao
Zhao, Zhihe
Liu, Jun
author_facet Li, Qiwen
Luo, Tao
Lu, Wenxin
Yi, Xiaoxiao
Zhao, Zhihe
Liu, Jun
author_sort Li, Qiwen
collection PubMed
description BACKGROUND: The periodontal ligament is essential for homeostasis of periodontal tissue. A hypoxic milieu of the periodontal tissue is generated under periodontitis or during orthodontic treatment, which affects the periodontal and bone remodelling process. Here, we provide a comprehensive proteomic characterization of periodontal ligament cells under hypoxic conditions, aiming to reveal previously unappreciated biological changes and to help advance hypoxia-based therapeutic strategies for periodontal diseases. METHODS: Human periodontal ligament cells (hPDLCs) were characterized using immunohistochemistry (IHC) and flow cytometry (FACS). Successful hypoxia treatment of hPDLCs with 1% O(2) was confirmed by qRT-PCR. Proliferation was evaluated using an MTT assay. The proteomic expression profile under hypoxia was studied with the isobaric tags for relative and absolute quantification (iTRAQ) approach followed by protein identification and bioinformatic analysis, and western blot verification was performed. RESULTS: The hPDLCs were positive for vimentin, CD73 and CD105 and negative for keratin, CD34 and CD45. After hypoxia treatment, the mRNA expression of hypoxia-inducible factor 1a (HIF1a) was upregulated. The proliferation rate was elevated during the first 6 h but decreased from 6 h to 72 h. A total of 220 differentially expressed proteins were quantified in hPDLCs under hypoxia (1% O(2), 24 h), including 153 upregulated and 67 downregulated proteins, five of which were verified by western blot analysis. The Gene Ontology enriched terms included the energy metabolic process, membrane-bound organelle and vesicle, and protein binding terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated several involved pathways, including glycolysis/gluconeogenesis, biosynthesis of amino acids, the HIF-1 signalling pathway, and focal adhesion. A protein–protein interaction (PPI) network demonstrated the dominant role of autophagy over apoptosis under hypoxia. CONCLUSION: The proteomic profile of hPDLCs under hypoxia was mainly related to energy metabolism, autophagy, and responses to stimuli such as adhesion and inflammation. Previously unrecognized proteins including solute carrier family proteins, heat shock proteins, ubiquitination-related enzymes, collagen and S100 family proteins are involved in adaptive response to hypoxia in hPDLCs and are thus of great research interest in future work. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12953-019-0151-2) contains supplementary material, which is available to authorized users.
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spelling pubmed-67176482019-09-06 Proteomic analysis of human periodontal ligament cells under hypoxia Li, Qiwen Luo, Tao Lu, Wenxin Yi, Xiaoxiao Zhao, Zhihe Liu, Jun Proteome Sci Research BACKGROUND: The periodontal ligament is essential for homeostasis of periodontal tissue. A hypoxic milieu of the periodontal tissue is generated under periodontitis or during orthodontic treatment, which affects the periodontal and bone remodelling process. Here, we provide a comprehensive proteomic characterization of periodontal ligament cells under hypoxic conditions, aiming to reveal previously unappreciated biological changes and to help advance hypoxia-based therapeutic strategies for periodontal diseases. METHODS: Human periodontal ligament cells (hPDLCs) were characterized using immunohistochemistry (IHC) and flow cytometry (FACS). Successful hypoxia treatment of hPDLCs with 1% O(2) was confirmed by qRT-PCR. Proliferation was evaluated using an MTT assay. The proteomic expression profile under hypoxia was studied with the isobaric tags for relative and absolute quantification (iTRAQ) approach followed by protein identification and bioinformatic analysis, and western blot verification was performed. RESULTS: The hPDLCs were positive for vimentin, CD73 and CD105 and negative for keratin, CD34 and CD45. After hypoxia treatment, the mRNA expression of hypoxia-inducible factor 1a (HIF1a) was upregulated. The proliferation rate was elevated during the first 6 h but decreased from 6 h to 72 h. A total of 220 differentially expressed proteins were quantified in hPDLCs under hypoxia (1% O(2), 24 h), including 153 upregulated and 67 downregulated proteins, five of which were verified by western blot analysis. The Gene Ontology enriched terms included the energy metabolic process, membrane-bound organelle and vesicle, and protein binding terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated several involved pathways, including glycolysis/gluconeogenesis, biosynthesis of amino acids, the HIF-1 signalling pathway, and focal adhesion. A protein–protein interaction (PPI) network demonstrated the dominant role of autophagy over apoptosis under hypoxia. CONCLUSION: The proteomic profile of hPDLCs under hypoxia was mainly related to energy metabolism, autophagy, and responses to stimuli such as adhesion and inflammation. Previously unrecognized proteins including solute carrier family proteins, heat shock proteins, ubiquitination-related enzymes, collagen and S100 family proteins are involved in adaptive response to hypoxia in hPDLCs and are thus of great research interest in future work. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12953-019-0151-2) contains supplementary material, which is available to authorized users. BioMed Central 2019-08-31 /pmc/articles/PMC6717648/ /pubmed/31496921 http://dx.doi.org/10.1186/s12953-019-0151-2 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Li, Qiwen
Luo, Tao
Lu, Wenxin
Yi, Xiaoxiao
Zhao, Zhihe
Liu, Jun
Proteomic analysis of human periodontal ligament cells under hypoxia
title Proteomic analysis of human periodontal ligament cells under hypoxia
title_full Proteomic analysis of human periodontal ligament cells under hypoxia
title_fullStr Proteomic analysis of human periodontal ligament cells under hypoxia
title_full_unstemmed Proteomic analysis of human periodontal ligament cells under hypoxia
title_short Proteomic analysis of human periodontal ligament cells under hypoxia
title_sort proteomic analysis of human periodontal ligament cells under hypoxia
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6717648/
https://www.ncbi.nlm.nih.gov/pubmed/31496921
http://dx.doi.org/10.1186/s12953-019-0151-2
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