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Upregulation of human GD3 synthase (hST8Sia I) gene expression during serum starvation-induced osteoblastic differentiation of MG-63 cells

In this study, we have firstly elucidated that serum starvation augmented the levels of human GD3 synthase (hST8Sia I) gene and ganglioside GD3 expression as well as bone morphogenic protein-2 and osteocalcin expression during MG-63 cell differentiation using RT-PCR, qPCR, Western blot and immunoflu...

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Autores principales: An, So-Young, Yoon, Hyun-Kyoung, Kim, Kyoung-Sook, Kim, Hee-Do, Cho, Jong-Hyun, Kim, Hyeon-Jun, Kim, Cheorl-Ho, Lee, Young-Choon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10621931/
https://www.ncbi.nlm.nih.gov/pubmed/37917776
http://dx.doi.org/10.1371/journal.pone.0293321
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author An, So-Young
Yoon, Hyun-Kyoung
Kim, Kyoung-Sook
Kim, Hee-Do
Cho, Jong-Hyun
Kim, Hyeon-Jun
Kim, Cheorl-Ho
Lee, Young-Choon
author_facet An, So-Young
Yoon, Hyun-Kyoung
Kim, Kyoung-Sook
Kim, Hee-Do
Cho, Jong-Hyun
Kim, Hyeon-Jun
Kim, Cheorl-Ho
Lee, Young-Choon
author_sort An, So-Young
collection PubMed
description In this study, we have firstly elucidated that serum starvation augmented the levels of human GD3 synthase (hST8Sia I) gene and ganglioside GD3 expression as well as bone morphogenic protein-2 and osteocalcin expression during MG-63 cell differentiation using RT-PCR, qPCR, Western blot and immunofluorescence microscopy. To evaluate upregulation of hST8Sia I gene during MG-63 cell differentiation by serum starvation, promoter area of the hST8Sia I gene was functionally analyzed. Promoter analysis using luciferase reporter assay system harboring various constructs of the hST8Sia I gene proved that the cis-acting region at -1146/-646, which includes binding sites of the known transcription factors AP-1, CREB, c-Ets-1 and NF-κB, displays the highest level of promoter activity in response to serum starvation in MG-63 cells. The -731/-722 region, which contains the NF-κB binding site, was proved to be essential for expression of the hST8Sia I gene by serum starvation in MG-63 cells by site-directed mutagenesis, NF-κB inhibition, and chromatin immunoprecipitation (ChIP) assay. Knockdown of hST8Sia I using shRNA suggested that expressions of hST8Sia I and GD3 have no apparent effect on differentiation of MG-63 cells. Moreover, the transcriptional activation of hST8Sia I gene by serum starvation was strongly hindered by SB203580, a p38MAPK inhibitor in MG-63 cells. From these results, it has been suggested that transcription activity of hST8Sia I gene by serum starvation in human osteosarcoma MG-63 cells is regulated by p38MAPK/NF-κB signaling pathway.
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spelling pubmed-106219312023-11-03 Upregulation of human GD3 synthase (hST8Sia I) gene expression during serum starvation-induced osteoblastic differentiation of MG-63 cells An, So-Young Yoon, Hyun-Kyoung Kim, Kyoung-Sook Kim, Hee-Do Cho, Jong-Hyun Kim, Hyeon-Jun Kim, Cheorl-Ho Lee, Young-Choon PLoS One Research Article In this study, we have firstly elucidated that serum starvation augmented the levels of human GD3 synthase (hST8Sia I) gene and ganglioside GD3 expression as well as bone morphogenic protein-2 and osteocalcin expression during MG-63 cell differentiation using RT-PCR, qPCR, Western blot and immunofluorescence microscopy. To evaluate upregulation of hST8Sia I gene during MG-63 cell differentiation by serum starvation, promoter area of the hST8Sia I gene was functionally analyzed. Promoter analysis using luciferase reporter assay system harboring various constructs of the hST8Sia I gene proved that the cis-acting region at -1146/-646, which includes binding sites of the known transcription factors AP-1, CREB, c-Ets-1 and NF-κB, displays the highest level of promoter activity in response to serum starvation in MG-63 cells. The -731/-722 region, which contains the NF-κB binding site, was proved to be essential for expression of the hST8Sia I gene by serum starvation in MG-63 cells by site-directed mutagenesis, NF-κB inhibition, and chromatin immunoprecipitation (ChIP) assay. Knockdown of hST8Sia I using shRNA suggested that expressions of hST8Sia I and GD3 have no apparent effect on differentiation of MG-63 cells. Moreover, the transcriptional activation of hST8Sia I gene by serum starvation was strongly hindered by SB203580, a p38MAPK inhibitor in MG-63 cells. From these results, it has been suggested that transcription activity of hST8Sia I gene by serum starvation in human osteosarcoma MG-63 cells is regulated by p38MAPK/NF-κB signaling pathway. Public Library of Science 2023-11-02 /pmc/articles/PMC10621931/ /pubmed/37917776 http://dx.doi.org/10.1371/journal.pone.0293321 Text en © 2023 An et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
An, So-Young
Yoon, Hyun-Kyoung
Kim, Kyoung-Sook
Kim, Hee-Do
Cho, Jong-Hyun
Kim, Hyeon-Jun
Kim, Cheorl-Ho
Lee, Young-Choon
Upregulation of human GD3 synthase (hST8Sia I) gene expression during serum starvation-induced osteoblastic differentiation of MG-63 cells
title Upregulation of human GD3 synthase (hST8Sia I) gene expression during serum starvation-induced osteoblastic differentiation of MG-63 cells
title_full Upregulation of human GD3 synthase (hST8Sia I) gene expression during serum starvation-induced osteoblastic differentiation of MG-63 cells
title_fullStr Upregulation of human GD3 synthase (hST8Sia I) gene expression during serum starvation-induced osteoblastic differentiation of MG-63 cells
title_full_unstemmed Upregulation of human GD3 synthase (hST8Sia I) gene expression during serum starvation-induced osteoblastic differentiation of MG-63 cells
title_short Upregulation of human GD3 synthase (hST8Sia I) gene expression during serum starvation-induced osteoblastic differentiation of MG-63 cells
title_sort upregulation of human gd3 synthase (hst8sia i) gene expression during serum starvation-induced osteoblastic differentiation of mg-63 cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10621931/
https://www.ncbi.nlm.nih.gov/pubmed/37917776
http://dx.doi.org/10.1371/journal.pone.0293321
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