Cargando…

Combined Effects of Androgen and Growth Hormone on Osteoblast Marker Expression in Mouse C2C12 and MC3T3-E1 Cells Induced by Bone Morphogenetic Protein

Osteoblasts undergo differentiation in response to various factors, including growth factors and steroids. Bone mass is diminished in androgen- and/or growth hormone (GH)-deficient patients. However the functional relationship between androgen and GH, and their combined effects on bone metabolism, r...

Descripción completa

Detalles Bibliográficos
Autores principales: Kimura, Kosuke, Terasaka, Tomohiro, Iwata, Nahoko, Katsuyama, Takayuki, Komatsubara, Motoshi, Nagao, Ryota, Inagaki, Kenichi, Otsuka, Fumio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5294959/
https://www.ncbi.nlm.nih.gov/pubmed/28067796
http://dx.doi.org/10.3390/jcm6010006
_version_ 1782505335939923968
author Kimura, Kosuke
Terasaka, Tomohiro
Iwata, Nahoko
Katsuyama, Takayuki
Komatsubara, Motoshi
Nagao, Ryota
Inagaki, Kenichi
Otsuka, Fumio
author_facet Kimura, Kosuke
Terasaka, Tomohiro
Iwata, Nahoko
Katsuyama, Takayuki
Komatsubara, Motoshi
Nagao, Ryota
Inagaki, Kenichi
Otsuka, Fumio
author_sort Kimura, Kosuke
collection PubMed
description Osteoblasts undergo differentiation in response to various factors, including growth factors and steroids. Bone mass is diminished in androgen- and/or growth hormone (GH)-deficient patients. However the functional relationship between androgen and GH, and their combined effects on bone metabolism, remains unclear. Here we investigated the mutual effects of androgen and GH on osteoblastic marker expression using mouse myoblastic C2C12 and osteoblast-like MC3T3-E1 cells. Combined treatment with dihydrotestosterone (DHT) and GH enhanced BMP-2-induced expression of Runx2, ALP, and osteocalcin mRNA, compared with the individual treatments in C2C12 cells. Co-treatment with DHT and GH activated Smad1/5/8 phosphorylation, Id-1 transcription, and ALP activity induced by BMP-2 in C2C12 cells but not in MC3T3-E1 cells. The insulin-like growth factor (IGF-I) mRNA level was amplified by GH and BMP-2 treatment and was restored by co-treatment with DHT in C2C12 cells. The mRNA level of the IGF-I receptor was not significantly altered by GH or DHT, while it was increased by IGF-I. In addition, IGF-I treatment increased collagen-1 mRNA expression, whereas blockage of endogenous IGF-I activity using an anti-IGF-I antibody failed to suppress the effect of GH and DHT on BMP-2-induced Runx2 expression in C2C12 cells, suggesting that endogenous IGF-I was not substantially involved in the underlying GH actions. On the other hand, androgen receptor and GH receptor mRNA expression was suppressed by BMP-2 in both cell lines, implying the existence of a feedback action. Collectively the results showed that the combined effects of androgen and GH facilitated BMP-2-induced osteoblast differentiation at an early stage by upregulating BMP receptor signaling.
format Online
Article
Text
id pubmed-5294959
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-52949592017-02-10 Combined Effects of Androgen and Growth Hormone on Osteoblast Marker Expression in Mouse C2C12 and MC3T3-E1 Cells Induced by Bone Morphogenetic Protein Kimura, Kosuke Terasaka, Tomohiro Iwata, Nahoko Katsuyama, Takayuki Komatsubara, Motoshi Nagao, Ryota Inagaki, Kenichi Otsuka, Fumio J Clin Med Article Osteoblasts undergo differentiation in response to various factors, including growth factors and steroids. Bone mass is diminished in androgen- and/or growth hormone (GH)-deficient patients. However the functional relationship between androgen and GH, and their combined effects on bone metabolism, remains unclear. Here we investigated the mutual effects of androgen and GH on osteoblastic marker expression using mouse myoblastic C2C12 and osteoblast-like MC3T3-E1 cells. Combined treatment with dihydrotestosterone (DHT) and GH enhanced BMP-2-induced expression of Runx2, ALP, and osteocalcin mRNA, compared with the individual treatments in C2C12 cells. Co-treatment with DHT and GH activated Smad1/5/8 phosphorylation, Id-1 transcription, and ALP activity induced by BMP-2 in C2C12 cells but not in MC3T3-E1 cells. The insulin-like growth factor (IGF-I) mRNA level was amplified by GH and BMP-2 treatment and was restored by co-treatment with DHT in C2C12 cells. The mRNA level of the IGF-I receptor was not significantly altered by GH or DHT, while it was increased by IGF-I. In addition, IGF-I treatment increased collagen-1 mRNA expression, whereas blockage of endogenous IGF-I activity using an anti-IGF-I antibody failed to suppress the effect of GH and DHT on BMP-2-induced Runx2 expression in C2C12 cells, suggesting that endogenous IGF-I was not substantially involved in the underlying GH actions. On the other hand, androgen receptor and GH receptor mRNA expression was suppressed by BMP-2 in both cell lines, implying the existence of a feedback action. Collectively the results showed that the combined effects of androgen and GH facilitated BMP-2-induced osteoblast differentiation at an early stage by upregulating BMP receptor signaling. MDPI 2017-01-05 /pmc/articles/PMC5294959/ /pubmed/28067796 http://dx.doi.org/10.3390/jcm6010006 Text en © 2017 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kimura, Kosuke
Terasaka, Tomohiro
Iwata, Nahoko
Katsuyama, Takayuki
Komatsubara, Motoshi
Nagao, Ryota
Inagaki, Kenichi
Otsuka, Fumio
Combined Effects of Androgen and Growth Hormone on Osteoblast Marker Expression in Mouse C2C12 and MC3T3-E1 Cells Induced by Bone Morphogenetic Protein
title Combined Effects of Androgen and Growth Hormone on Osteoblast Marker Expression in Mouse C2C12 and MC3T3-E1 Cells Induced by Bone Morphogenetic Protein
title_full Combined Effects of Androgen and Growth Hormone on Osteoblast Marker Expression in Mouse C2C12 and MC3T3-E1 Cells Induced by Bone Morphogenetic Protein
title_fullStr Combined Effects of Androgen and Growth Hormone on Osteoblast Marker Expression in Mouse C2C12 and MC3T3-E1 Cells Induced by Bone Morphogenetic Protein
title_full_unstemmed Combined Effects of Androgen and Growth Hormone on Osteoblast Marker Expression in Mouse C2C12 and MC3T3-E1 Cells Induced by Bone Morphogenetic Protein
title_short Combined Effects of Androgen and Growth Hormone on Osteoblast Marker Expression in Mouse C2C12 and MC3T3-E1 Cells Induced by Bone Morphogenetic Protein
title_sort combined effects of androgen and growth hormone on osteoblast marker expression in mouse c2c12 and mc3t3-e1 cells induced by bone morphogenetic protein
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5294959/
https://www.ncbi.nlm.nih.gov/pubmed/28067796
http://dx.doi.org/10.3390/jcm6010006
work_keys_str_mv AT kimurakosuke combinedeffectsofandrogenandgrowthhormoneonosteoblastmarkerexpressioninmousec2c12andmc3t3e1cellsinducedbybonemorphogeneticprotein
AT terasakatomohiro combinedeffectsofandrogenandgrowthhormoneonosteoblastmarkerexpressioninmousec2c12andmc3t3e1cellsinducedbybonemorphogeneticprotein
AT iwatanahoko combinedeffectsofandrogenandgrowthhormoneonosteoblastmarkerexpressioninmousec2c12andmc3t3e1cellsinducedbybonemorphogeneticprotein
AT katsuyamatakayuki combinedeffectsofandrogenandgrowthhormoneonosteoblastmarkerexpressioninmousec2c12andmc3t3e1cellsinducedbybonemorphogeneticprotein
AT komatsubaramotoshi combinedeffectsofandrogenandgrowthhormoneonosteoblastmarkerexpressioninmousec2c12andmc3t3e1cellsinducedbybonemorphogeneticprotein
AT nagaoryota combinedeffectsofandrogenandgrowthhormoneonosteoblastmarkerexpressioninmousec2c12andmc3t3e1cellsinducedbybonemorphogeneticprotein
AT inagakikenichi combinedeffectsofandrogenandgrowthhormoneonosteoblastmarkerexpressioninmousec2c12andmc3t3e1cellsinducedbybonemorphogeneticprotein
AT otsukafumio combinedeffectsofandrogenandgrowthhormoneonosteoblastmarkerexpressioninmousec2c12andmc3t3e1cellsinducedbybonemorphogeneticprotein