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The expression and role of serotonin receptor 5HTR2A in canine osteoblasts and an osteosarcoma cell line

BACKGROUND: The significance of the serotonergic system in bone physiology and, more specifically, the importance of the five hydroxytryptamine receptor 2A (5HTR2A) in normal osteoblast proliferation have been previously described; however the role of serotonin in osteosarcoma remains unclear. Parti...

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Autores principales: Bracha, Shay, Viall, Austin, Goodall, Cheri, Stang, Bernadette, Ruaux, Craig, Seguin, Bernard, Chappell, Patrick E
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3895671/
https://www.ncbi.nlm.nih.gov/pubmed/24330646
http://dx.doi.org/10.1186/1746-6148-9-251
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author Bracha, Shay
Viall, Austin
Goodall, Cheri
Stang, Bernadette
Ruaux, Craig
Seguin, Bernard
Chappell, Patrick E
author_facet Bracha, Shay
Viall, Austin
Goodall, Cheri
Stang, Bernadette
Ruaux, Craig
Seguin, Bernard
Chappell, Patrick E
author_sort Bracha, Shay
collection PubMed
description BACKGROUND: The significance of the serotonergic system in bone physiology and, more specifically, the importance of the five hydroxytryptamine receptor 2A (5HTR2A) in normal osteoblast proliferation have been previously described; however the role of serotonin in osteosarcoma remains unclear. Particularly, the expression and function of 5HTR2A in canine osteosarcoma has not yet been studied, thus we sought to determine if this indoleamine modulates cellular proliferation in vitro. Using real time quantitative reverse transcription PCR and immunoblot analyses, we explored receptor expression and signaling differences between non-neoplastic canine osteoblasts (CnOb) and an osteosarcoma cell line (COS). To elucidate specific serotonergic signaling pathways triggered by 5HTR2A, we performed immunoblots for ERK and CREB. Finally, we compared cell viability and the induction of apoptosis in the presence 5HTR2A agonists and antagonists. RESULTS: 5HTR2A was overexpressed in the malignant cell line in comparison to normal cells. In CnOb cells, ERK phosphorylation (ERK-P) decreased in response to both serotonin and a specific 5HTR2A antagonist, ritanserin. In contrast, ERK-P abundance increased in COS cells following either treatment. While endogenous CREB was undetectable in CnOb, CREB was observed constitutively in COS, with expression and exhibited increased CREB phosphorylation following escalating concentrations of ritanserin. To determine the influence of 5HTR2A signaling on cell viability we challenged cells with ritanserin and serotonin. Our findings confirmed that serotonin treatment promoted cell viability in malignant cells but not in normal osteoblasts. Conversely, ritanserin reduced cell viability in both the normal and osteosarcoma cells. Further, ritanserin induced apoptosis in COS at the same concentrations associated with decreased cell viability. CONCLUSIONS: These findings confirm the existence of a functional 5HTR2A in a canine osteosarcoma cell line. Results indicate that intracellular second messenger signal coupling of 5HTR2A is different between normal and malignant cells, warranting further research to investigate its potential as a novel therapeutic target for canine osteosarcoma.
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spelling pubmed-38956712014-01-21 The expression and role of serotonin receptor 5HTR2A in canine osteoblasts and an osteosarcoma cell line Bracha, Shay Viall, Austin Goodall, Cheri Stang, Bernadette Ruaux, Craig Seguin, Bernard Chappell, Patrick E BMC Vet Res Research Article BACKGROUND: The significance of the serotonergic system in bone physiology and, more specifically, the importance of the five hydroxytryptamine receptor 2A (5HTR2A) in normal osteoblast proliferation have been previously described; however the role of serotonin in osteosarcoma remains unclear. Particularly, the expression and function of 5HTR2A in canine osteosarcoma has not yet been studied, thus we sought to determine if this indoleamine modulates cellular proliferation in vitro. Using real time quantitative reverse transcription PCR and immunoblot analyses, we explored receptor expression and signaling differences between non-neoplastic canine osteoblasts (CnOb) and an osteosarcoma cell line (COS). To elucidate specific serotonergic signaling pathways triggered by 5HTR2A, we performed immunoblots for ERK and CREB. Finally, we compared cell viability and the induction of apoptosis in the presence 5HTR2A agonists and antagonists. RESULTS: 5HTR2A was overexpressed in the malignant cell line in comparison to normal cells. In CnOb cells, ERK phosphorylation (ERK-P) decreased in response to both serotonin and a specific 5HTR2A antagonist, ritanserin. In contrast, ERK-P abundance increased in COS cells following either treatment. While endogenous CREB was undetectable in CnOb, CREB was observed constitutively in COS, with expression and exhibited increased CREB phosphorylation following escalating concentrations of ritanserin. To determine the influence of 5HTR2A signaling on cell viability we challenged cells with ritanserin and serotonin. Our findings confirmed that serotonin treatment promoted cell viability in malignant cells but not in normal osteoblasts. Conversely, ritanserin reduced cell viability in both the normal and osteosarcoma cells. Further, ritanserin induced apoptosis in COS at the same concentrations associated with decreased cell viability. CONCLUSIONS: These findings confirm the existence of a functional 5HTR2A in a canine osteosarcoma cell line. Results indicate that intracellular second messenger signal coupling of 5HTR2A is different between normal and malignant cells, warranting further research to investigate its potential as a novel therapeutic target for canine osteosarcoma. BioMed Central 2013-12-12 /pmc/articles/PMC3895671/ /pubmed/24330646 http://dx.doi.org/10.1186/1746-6148-9-251 Text en Copyright © 2013 Bracha et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Bracha, Shay
Viall, Austin
Goodall, Cheri
Stang, Bernadette
Ruaux, Craig
Seguin, Bernard
Chappell, Patrick E
The expression and role of serotonin receptor 5HTR2A in canine osteoblasts and an osteosarcoma cell line
title The expression and role of serotonin receptor 5HTR2A in canine osteoblasts and an osteosarcoma cell line
title_full The expression and role of serotonin receptor 5HTR2A in canine osteoblasts and an osteosarcoma cell line
title_fullStr The expression and role of serotonin receptor 5HTR2A in canine osteoblasts and an osteosarcoma cell line
title_full_unstemmed The expression and role of serotonin receptor 5HTR2A in canine osteoblasts and an osteosarcoma cell line
title_short The expression and role of serotonin receptor 5HTR2A in canine osteoblasts and an osteosarcoma cell line
title_sort expression and role of serotonin receptor 5htr2a in canine osteoblasts and an osteosarcoma cell line
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3895671/
https://www.ncbi.nlm.nih.gov/pubmed/24330646
http://dx.doi.org/10.1186/1746-6148-9-251
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