Differentiation-inducing factor-1 alters canonical Wnt signaling and suppresses alkaline phosphatase expression in osteoblast-like cell lines

Etsuko Matsuzaki, Fumi Takahashi-Yanaga, Yoshikazu Miwa, Masato Hirata, Yutaka Watanabe, Noriharu Sato, Sachio Morimoto, Takao Hirofuji, Katsumasa Maeda, Toshiyuki Sasaguri

研究成果: ジャーナルへの寄稿記事

31 引用 (Scopus)

抄録

Because DIF-1 has been shown to affect Wnt/β-catenin signaling pathway, the effects of DIF-1 on osteoblast-like cell lines, SaOS-2 and MC3T3-E1, were examined. We found that DIF-1 inhibited this pathway, resulting in the suppression of ALP promoter activity through the TCF/LEF binding site. Introduction: Differentiation-inducing factor-1 (DIF-1), a morphogen of Dictyostelium, inhibits cell proliferation and induces cell differentiation in several mammalian cells. Previous studies showed that DIF-1 activated glycogen synthase kinase-3β, suggesting that this chemical could affect the Wnt/β-catenin signaling pathway. This pathway has been shown to be involved in bone biology. Materials and Methods: We studied the effects of DIF-1 on SaOS-2 and MC3T3-E1, osteosarcoma cell lines widely used as a model system for ostoblastic cells and murine osteoblast-like cell line, respectively. Reporter gene assays were also carried out to examine the effect of DIF-1 on the Wnt/β-catenin signaling pathway. Results: DIF-1 inhibited SaOS-2 proliferation and reduced alkaline phosphatase (ALP) activity in a concentration-and a time-dependent manner. The expression of ALP was markedly suppressed by DIF-1-treatment in protein and mRNA levels. DIF-1 also suppressed the expression of other osteoblast differentiation markers, including core binding factor α1, type I collagen, and osteocalcin, in protein and mRNA levels and inhibited osteoblast-mediated mineralization. Subsequently, we examined the effect of DIF-1 on the Wnt/β-catenin signaling pathway. We found that DIF-1 suppressed the expression of β-catenin protein and the activity of the reporter gene containing T-cell factor/lymphoid enhancer-binding factor (TCF/LEF) consensus binding sites. We examined the effect of DIF-1 on a reporter gene driven by the human ALP promoter and found that DIF-1 significantly reduced the ALP reporter gene activity through the TCF/LEF binding site (-1023/-1017 bp). Furthermore, the effect of DIF-1 on MC3T3-E1, a murine osteoblast-like cell line, was examined, and it was found that DIF-1 suppressed ALP mRNA expression by the reduction of the ALP reporter gene activity through the TCF/LEF binding site. Conclusions: Our data suggest that DIF-1 inhibits Wnt/β-catenin signaling, resulting in the suppression of ALP promoter activity. To our knowledge, this is the first report to analyze the role of the TCF/LEF binding site (-1023/-1017 bp) of the ALP gene promoter in osteoblast-like cell lines.

元の言語英語
ページ(範囲)1307-1316
ページ数10
ジャーナルJournal of Bone and Mineral Research
21
発行部数8
DOI
出版物ステータス出版済み - 8 1 2006

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Osteoblasts
Alkaline Phosphatase
TCF Transcription Factors
Cell Line
Catenins
Reporter Genes
Wnt Signaling Pathway
Binding Sites
1-((3,5-dichloro)-2,6-dihydroxy-4-methoxyphenyl)-1-hexanone
Messenger RNA
Core Binding Factors
Glycogen Synthase Kinase 3
Dictyostelium
Osteocalcin
Differentiation Antigens
Osteosarcoma
Collagen Type I
Cell Differentiation

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Orthopedics and Sports Medicine

これを引用

Differentiation-inducing factor-1 alters canonical Wnt signaling and suppresses alkaline phosphatase expression in osteoblast-like cell lines. / Matsuzaki, Etsuko; Takahashi-Yanaga, Fumi; Miwa, Yoshikazu; Hirata, Masato; Watanabe, Yutaka; Sato, Noriharu; Morimoto, Sachio; Hirofuji, Takao; Maeda, Katsumasa; Sasaguri, Toshiyuki.

:: Journal of Bone and Mineral Research, 巻 21, 番号 8, 01.08.2006, p. 1307-1316.

研究成果: ジャーナルへの寄稿記事

Matsuzaki, E, Takahashi-Yanaga, F, Miwa, Y, Hirata, M, Watanabe, Y, Sato, N, Morimoto, S, Hirofuji, T, Maeda, K & Sasaguri, T 2006, 'Differentiation-inducing factor-1 alters canonical Wnt signaling and suppresses alkaline phosphatase expression in osteoblast-like cell lines', Journal of Bone and Mineral Research, 巻. 21, 番号 8, pp. 1307-1316. https://doi.org/10.1359/jbmr.060512
Matsuzaki, Etsuko ; Takahashi-Yanaga, Fumi ; Miwa, Yoshikazu ; Hirata, Masato ; Watanabe, Yutaka ; Sato, Noriharu ; Morimoto, Sachio ; Hirofuji, Takao ; Maeda, Katsumasa ; Sasaguri, Toshiyuki. / Differentiation-inducing factor-1 alters canonical Wnt signaling and suppresses alkaline phosphatase expression in osteoblast-like cell lines. :: Journal of Bone and Mineral Research. 2006 ; 巻 21, 番号 8. pp. 1307-1316.
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abstract = "Because DIF-1 has been shown to affect Wnt/β-catenin signaling pathway, the effects of DIF-1 on osteoblast-like cell lines, SaOS-2 and MC3T3-E1, were examined. We found that DIF-1 inhibited this pathway, resulting in the suppression of ALP promoter activity through the TCF/LEF binding site. Introduction: Differentiation-inducing factor-1 (DIF-1), a morphogen of Dictyostelium, inhibits cell proliferation and induces cell differentiation in several mammalian cells. Previous studies showed that DIF-1 activated glycogen synthase kinase-3β, suggesting that this chemical could affect the Wnt/β-catenin signaling pathway. This pathway has been shown to be involved in bone biology. Materials and Methods: We studied the effects of DIF-1 on SaOS-2 and MC3T3-E1, osteosarcoma cell lines widely used as a model system for ostoblastic cells and murine osteoblast-like cell line, respectively. Reporter gene assays were also carried out to examine the effect of DIF-1 on the Wnt/β-catenin signaling pathway. Results: DIF-1 inhibited SaOS-2 proliferation and reduced alkaline phosphatase (ALP) activity in a concentration-and a time-dependent manner. The expression of ALP was markedly suppressed by DIF-1-treatment in protein and mRNA levels. DIF-1 also suppressed the expression of other osteoblast differentiation markers, including core binding factor α1, type I collagen, and osteocalcin, in protein and mRNA levels and inhibited osteoblast-mediated mineralization. Subsequently, we examined the effect of DIF-1 on the Wnt/β-catenin signaling pathway. We found that DIF-1 suppressed the expression of β-catenin protein and the activity of the reporter gene containing T-cell factor/lymphoid enhancer-binding factor (TCF/LEF) consensus binding sites. We examined the effect of DIF-1 on a reporter gene driven by the human ALP promoter and found that DIF-1 significantly reduced the ALP reporter gene activity through the TCF/LEF binding site (-1023/-1017 bp). Furthermore, the effect of DIF-1 on MC3T3-E1, a murine osteoblast-like cell line, was examined, and it was found that DIF-1 suppressed ALP mRNA expression by the reduction of the ALP reporter gene activity through the TCF/LEF binding site. Conclusions: Our data suggest that DIF-1 inhibits Wnt/β-catenin signaling, resulting in the suppression of ALP promoter activity. To our knowledge, this is the first report to analyze the role of the TCF/LEF binding site (-1023/-1017 bp) of the ALP gene promoter in osteoblast-like cell lines.",
author = "Etsuko Matsuzaki and Fumi Takahashi-Yanaga and Yoshikazu Miwa and Masato Hirata and Yutaka Watanabe and Noriharu Sato and Sachio Morimoto and Takao Hirofuji and Katsumasa Maeda and Toshiyuki Sasaguri",
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T1 - Differentiation-inducing factor-1 alters canonical Wnt signaling and suppresses alkaline phosphatase expression in osteoblast-like cell lines

AU - Matsuzaki, Etsuko

AU - Takahashi-Yanaga, Fumi

AU - Miwa, Yoshikazu

AU - Hirata, Masato

AU - Watanabe, Yutaka

AU - Sato, Noriharu

AU - Morimoto, Sachio

AU - Hirofuji, Takao

AU - Maeda, Katsumasa

AU - Sasaguri, Toshiyuki

PY - 2006/8/1

Y1 - 2006/8/1

N2 - Because DIF-1 has been shown to affect Wnt/β-catenin signaling pathway, the effects of DIF-1 on osteoblast-like cell lines, SaOS-2 and MC3T3-E1, were examined. We found that DIF-1 inhibited this pathway, resulting in the suppression of ALP promoter activity through the TCF/LEF binding site. Introduction: Differentiation-inducing factor-1 (DIF-1), a morphogen of Dictyostelium, inhibits cell proliferation and induces cell differentiation in several mammalian cells. Previous studies showed that DIF-1 activated glycogen synthase kinase-3β, suggesting that this chemical could affect the Wnt/β-catenin signaling pathway. This pathway has been shown to be involved in bone biology. Materials and Methods: We studied the effects of DIF-1 on SaOS-2 and MC3T3-E1, osteosarcoma cell lines widely used as a model system for ostoblastic cells and murine osteoblast-like cell line, respectively. Reporter gene assays were also carried out to examine the effect of DIF-1 on the Wnt/β-catenin signaling pathway. Results: DIF-1 inhibited SaOS-2 proliferation and reduced alkaline phosphatase (ALP) activity in a concentration-and a time-dependent manner. The expression of ALP was markedly suppressed by DIF-1-treatment in protein and mRNA levels. DIF-1 also suppressed the expression of other osteoblast differentiation markers, including core binding factor α1, type I collagen, and osteocalcin, in protein and mRNA levels and inhibited osteoblast-mediated mineralization. Subsequently, we examined the effect of DIF-1 on the Wnt/β-catenin signaling pathway. We found that DIF-1 suppressed the expression of β-catenin protein and the activity of the reporter gene containing T-cell factor/lymphoid enhancer-binding factor (TCF/LEF) consensus binding sites. We examined the effect of DIF-1 on a reporter gene driven by the human ALP promoter and found that DIF-1 significantly reduced the ALP reporter gene activity through the TCF/LEF binding site (-1023/-1017 bp). Furthermore, the effect of DIF-1 on MC3T3-E1, a murine osteoblast-like cell line, was examined, and it was found that DIF-1 suppressed ALP mRNA expression by the reduction of the ALP reporter gene activity through the TCF/LEF binding site. Conclusions: Our data suggest that DIF-1 inhibits Wnt/β-catenin signaling, resulting in the suppression of ALP promoter activity. To our knowledge, this is the first report to analyze the role of the TCF/LEF binding site (-1023/-1017 bp) of the ALP gene promoter in osteoblast-like cell lines.

AB - Because DIF-1 has been shown to affect Wnt/β-catenin signaling pathway, the effects of DIF-1 on osteoblast-like cell lines, SaOS-2 and MC3T3-E1, were examined. We found that DIF-1 inhibited this pathway, resulting in the suppression of ALP promoter activity through the TCF/LEF binding site. Introduction: Differentiation-inducing factor-1 (DIF-1), a morphogen of Dictyostelium, inhibits cell proliferation and induces cell differentiation in several mammalian cells. Previous studies showed that DIF-1 activated glycogen synthase kinase-3β, suggesting that this chemical could affect the Wnt/β-catenin signaling pathway. This pathway has been shown to be involved in bone biology. Materials and Methods: We studied the effects of DIF-1 on SaOS-2 and MC3T3-E1, osteosarcoma cell lines widely used as a model system for ostoblastic cells and murine osteoblast-like cell line, respectively. Reporter gene assays were also carried out to examine the effect of DIF-1 on the Wnt/β-catenin signaling pathway. Results: DIF-1 inhibited SaOS-2 proliferation and reduced alkaline phosphatase (ALP) activity in a concentration-and a time-dependent manner. The expression of ALP was markedly suppressed by DIF-1-treatment in protein and mRNA levels. DIF-1 also suppressed the expression of other osteoblast differentiation markers, including core binding factor α1, type I collagen, and osteocalcin, in protein and mRNA levels and inhibited osteoblast-mediated mineralization. Subsequently, we examined the effect of DIF-1 on the Wnt/β-catenin signaling pathway. We found that DIF-1 suppressed the expression of β-catenin protein and the activity of the reporter gene containing T-cell factor/lymphoid enhancer-binding factor (TCF/LEF) consensus binding sites. We examined the effect of DIF-1 on a reporter gene driven by the human ALP promoter and found that DIF-1 significantly reduced the ALP reporter gene activity through the TCF/LEF binding site (-1023/-1017 bp). Furthermore, the effect of DIF-1 on MC3T3-E1, a murine osteoblast-like cell line, was examined, and it was found that DIF-1 suppressed ALP mRNA expression by the reduction of the ALP reporter gene activity through the TCF/LEF binding site. Conclusions: Our data suggest that DIF-1 inhibits Wnt/β-catenin signaling, resulting in the suppression of ALP promoter activity. To our knowledge, this is the first report to analyze the role of the TCF/LEF binding site (-1023/-1017 bp) of the ALP gene promoter in osteoblast-like cell lines.

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