CCAAT/Enhancer-binding Protein β Regulates the Repression of Type II Collagen Expression during the Differentiation from Proliferative to Hypertrophic Chondrocytes

Ushijima, Takahiro; Okazaki, Ken; Tsushima, Hiroyuki; Iwamoto, Yukihide

doi:10.1074/jbc.m113.492843

Cited by 34 publications

(35 citation statements)

References 42 publications

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“…The expression of C/EBPβ was increased by the infection of adenovirus vector expressing C/EBPβ-LAP, indicating that transfection of C/EBPβ was effectively performed. As we previously reported, hypertrophic transition of cultured tibias was observed in morphology as well as protein expression [19]. The expression of Ihh and RUNX2 was increased in the tibias which were transfected with C/EBPβ, suggesting that C/EBPβ regulates the expression of Ihh.…”

Section: Resultssupporting

confidence: 66%

“…Increase of the mRNA (not shown) and nuclear protein of C/EBPβ-LAP by infection of adenovirus vector demonstrated that transfection of C/EBPβ was effectively performed (Figure 2A). We previously reported that in the same model, exogenous C/EBPβ significantly increased the expression of Runx2 on the 4th and 7th days [19]. The expression of Ihh was significantly increased at all the differentiation stages (Figure 2B).…”

Section: Resultssupporting

confidence: 60%

“…Moreover, C/EBPβ stimulates the expression of MMP13 by interacting with RUNX2 during chondrocyte differentiation and OA development [17]. In fact, C/EBPβ increased the expression of RUNX2 in differentiating ATDC5 cells [19]. In the present study, therefore, we focused on the cooperative binding of C/EBPβ and RUNX2 to the C/EBPβ binding element in the Ihh promoter.…”

Section: Discussionmentioning

confidence: 94%

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CCAAT/Enhancer Binding Protein β Regulates Expression of Indian Hedgehog during Chondrocytes Differentiation

et al. 2014

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Background CCAAT/enhancer binding protein β (C/EBPβ) is a transcription factor that promotes hypertrophic differentiation of chondrocytes. Indian hedgehog (Ihh) also stimulates the hypertrophic transition of chondrocytes. Furthermore, runt-related transcription factor-2 (RUNX2) was reported to regulate chondrocyte maturation during skeletal development and to directly regulate transcriptional activity of Ihh. In this study, we investigated whether the interaction of C/EBPβ and RUNX2 regulates the expression of Ihh during chondrocyte differentiation.Methodology/ResultsImmunohistochemistry of embryonic growth plate revealed that both C/EBPβ and Ihh were strongly expressed in pre-hypertrophic and hypertrophic chondrocytes. Overexpression of C/EBPβ by adenovirus vector in ATDC5 cells caused marked stimulation of Ihh and Runx2. Conversely, knockdown of C/EBPβ by lentivirus expressing shRNA significantly repressed Ihh and Runx2 in ATDC5 cells. A reporter assay revealed that C/EBPβ stimulated transcriptional activity of Ihh. Deletion and mutation analysis showed that the C/EBPβ responsive element was located between −214 and −210 bp in the Ihh promoter. An electrophoretic mobility shift assay (EMSA) and a chromatin immunoprecipitation (ChIP) assay also revealed the direct binding of C/EBPβ to this region. Moreover, reporter assays demonstrated that RUNX2 failed to stimulate the transcriptional activity of the Ihh promoter harboring a mutation at the C/EBPβ binding site. EMSA and ChIP assays showed that RUNX2 interacted to this element with C/EBPβ. Immunoprecipitation revealed that RUNX2 and C/EBPβ formed heterodimer complex with each other in the nuclei of chondrocytes. These data suggested that the C/EBPβ binding element is also important for RUNX2 to regulate the expression of Ihh. Ex vivo organ culture of mouse limbs transfected with C/EBPβ showed that the expression of Ihh and RUNX2 was increased upon ectopic C/EBPβ expression.ConclusionsC/EBPβ and RUNX2 cooperatively stimulate expression of Ihh through direct interactions with a C/EBPβ binding element, which further promotes hypertrophic differentiation of chondrocytes during the chondrocyte differentiation process.

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Section: Resultssupporting

confidence: 66%

Section: Resultssupporting

confidence: 60%

Section: Discussionmentioning

confidence: 94%

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CCAAT/Enhancer Binding Protein β Regulates Expression of Indian Hedgehog during Chondrocytes Differentiation

et al. 2014

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“…For cartilage-specific genes, these include δEF1, AP-2α, Snail, Slug, Twist1, and C/EBPβ. [18][19][20][21][22] Among these, δEF1, AP-2α, and Twist1 are expressed by chondrocyte progenitors and maintain their undifferentiated state. 18,20,21,23 Their expression in the salivary gland was therefore compared with their expression in pleomorphic adenoma.…”

Section: Discussionmentioning

confidence: 99%

“…17 Negative regulators include δEF1, AP-2α, Snail, Slug, Twist1, and C/EBPβ. [18][19][20][21][22] Snail, Slug, and C/EBPβ are expressed by hypertrophic chondrocytes, which indicates that they control terminal chondrocyte differentiation and promote endochondral ossification; 11,19,22 δEF1, AP-2α, and Twist1 are expressed by chondroprogenitors, which indicates that they control early-phase chondrogenesis. 18,20,21,23 The aim of our study was to clarify the mechanisms behind chondrogenesis in pleomorphic adenoma epithelial cells.…”

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confidence: 99%

Transcription factors related to chondrogenesis in pleomorphic adenoma of the salivary gland: a mechanism of mesenchymal tissue formation

Matsumoto

Sato

Maeda

et al. 2016

Laboratory Investigation

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In salivary gland pleomorphic adenoma, expression of extracellular matrix (ECM) substances indicates that tumor epithelial cells are becoming chondrogenic and will produce cartilage-like mesenchymal tissues. Sox9, the master transcription factor of chondrogenesis, is expressed in mouse salivary gland cells. To clarify the mechanism behind chondrogenesis in tumor epithelial cells, we examined the expression of transcription factors related to chondrogenesis in tumors and salivary glands. Reverse transcriptase-polymerase chain reaction (RT-PCR), quantitative real-time RT-PCR, and immunostaining were performed on pleomorphic adenoma tissues, salivary gland tissues, and human submandibular gland (HSG) cells. The mRNAs of essential transcription factors for chondrogenesis-Sox9, Sox6, and Sox5-were detected in both tumor and salivary gland tissues. The mRNAs of aggrecan and type II collagen-cartilage-specific ECM substances-were detected only in tumors. Sox9 and Sox6 proteins were colocalized in many epithelial cells in tumors and salivary glands. Tumor epithelial cells also possessed aggrecan protein and occasionally type II collagen protein. Moreover, mRNAs for transcription repressors of chondrogenesis δEF1 and AP-2α were detected in both tumors and salivary glands, whereas Twist1 mRNA was detected only in salivary glands and was at significantly low-to-undetectable levels in tumors. Twist1 protein was localized in the Sox9-expressing salivary gland cells. HSG cells expressed Sox9, Sox6, and Twist1, but not aggrecan or type II collagen, and thus were similar to salivary gland cells. Twist1 depletion by Twist1 siRNA led to the upregulation of aggrecan and type II collagen mRNA expression in HSG cells. In contrast, forced expression of Twist1, using Twist1 cDNA, resulted in the downregulation of both these genes. Taken together, these results indicate that salivary gland cells have a potential for chondrogenesis, and Twist1 depletion concomitant with neoplastic transformation, which would permit tumor epithelial cells to produce cartilage-like mesenchymal tissues in salivary gland pleomorphic adenoma. (2016) 96, 16-24; doi:10.1038/labinvest.2015 published online 26 October 2015 Pleomorphic adenoma is the most common tumor of the salivary gland. It is characterized by mixed appearance of epithelial and mesenchymal components. 1,2 A number of ultrastructural and immunohistochemical studies have shown that the mesenchymal component (typically composed of myxoid and chondroid tissues) is epithelial in origin (e.g., Dardick et al. 3,4 and Erlandson et al. 5 ). Immunohistochemical and in situ hybridization studies have shown the production of aggrecan and type II collagen, extracellular matrix (ECM) substances specific to cartilage, in the myxoid and chondroid tissues. [6][7][8][9] As aggrecan is also produced in the epithelial component, 6,9 epithelial cells of pleomorphic adenoma could conceivably become chondrogenic and produce mesenchymal tissues similar to cartilage. Laboratory InvestigationWhen forced to express ...

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De Novo ARID1B mutations cause growth delay associated with aberrant Wnt/β–catenin signaling

Liu

et al. 2020

Human Mutation

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Haploinsufficiency of ARID1B (AT-rich interaction domain 1B) has been involved in autism spectrum disorder, nonsyndromic and syndromic intellectual disability, and corpus callosum agenesis. Growth impairment is a major clinical feature caused by ARID1B mutations; however, the mechanistic link has not been elucidated. Here, we confirm that growth delay is a common characteristic of patients with ARID1B mutations, which may be associated with dysregulation of the Wnt/β-catenin signaling pathway. An analysis of patients harboring pathogenic variants of ARID1B revealed that nearly half had short stature and nearly all had below-average height.Moreover, the percentage of patients with short stature increased with age.Knockdown of arid1b in zebrafish embryos markedly reduced body length and perturbed the expression of both chondrogenic and osteogenic genes including sox9a, col2a1a, runx2b, and col10a1. Knockout of Arid1b in chondrogenic ATDC5 cells inhibited chondrocyte proliferation and differentiation. Finally, Wnt/β-catenin signaling was perturbed in Arid1b-depleted zebrafish embryos and Arid1b knockout ATDC5 cells. These data indicate that ARID1B modulates bone growth possibly via regulation of the Wnt/β-catenin pathway, and may be an appropriate target for gene therapy in disorders of growth and development. K E Y W O R D SARID1B, bone growth, gene mutation, growth delay, short stature, Wnt/β-catenin

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CCAAT/Enhancer-binding Protein β Regulates the Repression of Type II Collagen Expression during the Differentiation from Proliferative to Hypertrophic Chondrocytes

Cited by 34 publications

References 42 publications

CCAAT/Enhancer Binding Protein β Regulates Expression of Indian Hedgehog during Chondrocytes Differentiation

CCAAT/Enhancer Binding Protein β Regulates Expression of Indian Hedgehog during Chondrocytes Differentiation

Transcription factors related to chondrogenesis in pleomorphic adenoma of the salivary gland: a mechanism of mesenchymal tissue formation

De Novo ARID1B mutations cause growth delay associated with aberrant Wnt/β–catenin signaling

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