Chromatin Immunoprecipitation on Microarray Analysis of Smad2/3 Binding Sites Reveals Roles of ETS1 and TFAP2A in Transforming Growth Factor β Signaling

Koinuma, Daizo; Tsutsumi, Sadami; Kamimura, Naoko; Taniguchi, Hirokazu; Miyazawa, Keiji; Sunamura, Makoto; Imamura, Takeshi; Miyazono, Kohei; Aburatani, Hiroyuki

doi:10.1128/mcb.01038-08

Cited by 182 publications

(187 citation statements)

References 57 publications

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“…53,54 In accordance with the down-regulation of the TGF-β pathway, we observed up-regulation of the Myc pathway that would also contribute to leukemogenesis and the global down-regulation of Smad2/3 targets as well as TGF-β receptors shown in Figure 6F. [56][57][58] Intriguingly, Zhou et al reported that SMAD2, a downstream mediator of TGF-β receptors, is constitutively activated and overexpressed in MDS BM precursors and that shRNA-mediated down-regulation or pharmacologic inhibition of TGF-β receptor 1 leads to enhanced hematopoiesis in a variety of MDS subtypes in vitro. 59 Thus, it is tempting to consider that the TGF-β signaling pathway inhibits leukemic transformation of MDS and that SETBP1 mutations contribute to the down-regulation of this important pathway.…”

Section: Discussionsupporting

confidence: 65%

SETBP1 mutations drive leukemic transformation of ASXL1-mutated MDS

Inoue

Matsui

Kitamura

2014

Experimental Hematology

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Section: Discussionsupporting

confidence: 65%

SETBP1 mutations drive leukemic transformation of ASXL1-mutated MDS

Inoue

Matsui

Kitamura

2014

Experimental Hematology

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“…19 GSEA revealed a significant overlap between the Fra-1-derived gene sets and the genes upregulated in EpRas cells upon TGF-β-induced EMT 21 (Table 1a). Genes associated with c-Myb 29 or with promoters occupied by Smad2/3, 30 two TFs connected to TGF-β and EMT, 31,32 were also enriched (Table 1b) Figure S6a, Figures 5a and b). Slug, encoded by snai2, was also increased (Figures 5a and b), whereas mRNA of snai1/Snail was moderately elevated and the protein undetectable (Figure 5a and data not shown).…”

Section: Resultsmentioning

confidence: 99%

Fra-1/AP-1 induces EMT in mammary epithelial cells by modulating Zeb1/2 and TGFβ expression

et al. 2014

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Epithelial-to-mesenchymal transition (EMT) is essential for embryonic morphogenesis and wound healing and critical for tumour cell invasion and dissemination. The AP-1 transcription factor Fra-1 has been implicated in tumorigenesis and in tumour-associated EMT in human breast cancer. We observed a significant inverse correlation between Fra-1 mRNA expression and distant-metastasis-free survival in a large cohort of breast cancer patients derived from multiple array data sets. This unique correlation among Fos genes prompted us to assess the evolutionary conservation between Fra-1 functions in EMT of human and mouse cells. Ectopic expression of Fra-1 in fully polarized, non-tumourigenic, mouse mammary epithelial EpH4 cells induced a mesenchymal phenotype, characterized by a loss of epithelial and gain of mesenchymal markers. Proliferation, motility and invasiveness were also increased in the resulting EpFra1 cells, and the cells were tumourigenic and efficiently colonized the lung upon transplantation. Molecular analyses revealed increased expression of Tgfβ1 and the EMT-inducing transcription factors Zeb1, Zeb2 and Slug. Mechanistically, Fra-1 binds to the tgfb1 and zeb2 promoters and to an evolutionarily conserved region in the first intron of zeb1. Furthermore, increased activity of a zeb2 promoter reporter was detected in EpFra1 cells and shown to depend on AP-1-binding sites. Inhibiting TGFβ signalling in EpFra1 cells moderately increased the expression of epithelial markers, whereas silencing of zeb1 or zeb2 restored the epithelial phenotype and decreased migration in vitro and tumorigenesis in vivo. Thus Fra-1 induces changes in the expression of genes encoding EMT-related transcription factors leading to the acquisition of mesenchymal, invasive and tumorigenic capacities by epithelial cells. This study defines a novel function of Fra-1/AP-1 in modulating tgfb1, zeb1 and zeb2 expression through direct binding to genomic regulatory regions, which establishes a basis for future in vivo genetic manipulations and preclinical studies using mouse models. Epithelial-to-mesenchymal transition (EMT) is a complex biological programme that occurs in physiological processes during embryonic development and wound healing as well as in pathological conditions, such as organ fibrosis and carcinogenesis. During EMT, cells lose epithelial features and acquire mesenchymal characteristics. The acquisition of a mesenchymal state by malignant cancer cells is associated with decreased cell-cell adhesion, and increased migratory and invasive properties, which are crucial for metastasis. [1][2][3][4][5] The adherens junction (AJ) protein E-cadherin, encoded by cdh1, is a central determinant of the epithelial state and its downregulation is the hallmark of EMT. A number of molecular pathways converging on E-cadherin have been implicated in EMT. Transcription factors (TF) of the Snail, Zeb and Twist families, initially identified as regulators of epithelialmesenchymal plasticity during morphogenesis were shown to orchestrate EMT by...

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“…MDA‐231‐D, Hs578T, and MCF7 cells were cultured in 10‐cm plates, and ChIP and ChIP‐seq were performed as described previously (Koinuma et al ., 2009; Murai et al ., 2015). Data were obtained using the Ion Proton sequencer (Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

ZEB1‐regulated inflammatory phenotype in breast cancer cells

et al. 2017

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Zinc finger E‐box binding protein 1 (ZEB1) and ZEB2 induce epithelial‐mesenchymal transition (EMT) and enhance cancer progression. However, the global view of transcriptional regulation by ZEB1 and ZEB2 is yet to be elucidated. Here, we identified a ZEB1‐regulated inflammatory phenotype in breast cancer cells using chromatin immunoprecipitation sequencing and RNA sequencing, followed by gene set enrichment analysis (GSEA) of ZEB1‐bound genes. Knockdown of ZEB1 and/or ZEB2 resulted in the downregulation of genes encoding inflammatory cytokines related to poor prognosis in patients with cancer, including IL6 and IL8, therefore suggesting that ZEB1 and ZEB2 have similar functions in terms of the regulation of production of inflammatory cytokines. Antibody array and ELISA experiments confirmed that ZEB1 controlled the production of the IL‐6 and IL‐8 proteins. The secretory proteins regulated by ZEB1 enhanced breast cancer cell proliferation and tumor growth. ZEB1 expression in breast cancer cells also affected the growth of fibroblasts in cell culture, and the accumulation of myeloid‐derived suppressor cells in tumors in vivo. These findings provide insight into the role of ZEB1 in the progression of cancer, mediated by inflammatory cytokines, along with the initiation of EMT.

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Chromatin Immunoprecipitation on Microarray Analysis of Smad2/3 Binding Sites Reveals Roles of ETS1 and TFAP2A in Transforming Growth Factor β Signaling

Cited by 182 publications

References 57 publications

SETBP1 mutations drive leukemic transformation of ASXL1-mutated MDS

SETBP1 mutations drive leukemic transformation of ASXL1-mutated MDS

Fra-1/AP-1 induces EMT in mammary epithelial cells by modulating Zeb1/2 and TGFβ expression

ZEB1‐regulated inflammatory phenotype in breast cancer cells

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