Diverse cellular and molecular mechanisms contribute to epithelial plasticity and metastasis

Grünert, Stefan; Jechlinger, Martin; Beug, Hartmut

doi:10.1038/nrm1175

Cited by 616 publications

(593 citation statements)

References 51 publications

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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).…”

Section: Resultsmentioning

confidence: 99%

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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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Section: Resultsmentioning

confidence: 99%

“…18 The phenotypic plasticity of this cell line upon Ras/MAPK pathway manipulation has been extensively documented. [19][20][21][22] Expression of c-Fos 18 or cJun 23 in EpH4 cells increases Fra-1 mRNA and promotes EMT. Here we show that ectopic expression of Fra-1 is sufficient to trigger a mesenchymal, invasive and tumorigenic programme in EpH4 cells.…”

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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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“…In this paper, we described an inducible cellular system (RafER-EpH4) to perform kinetic analysis of EMT, central for both development and tumor progression (Grunert et al, 2003). In this system, MAPK hyperactivation can be induced at will by 4HT activation of the fusion protein dRafER, its strength depending on 4HT concentration.…”

Section: Discussionmentioning

confidence: 99%

“…from benign adenoma to metastatic carcinoma). In particular, the ability of transforming growth factor b (TGFb) to induce EMT has been correlated with a tumor-promoting role of TGFb during late-stage cancer development, contrasted by TGFb-induced cell cycle arrest and apoptosis induced in normal epithelial cells (Derynck et al, 2002;Grunert et al, 2003). Using fully polarized mammary epithelial cells (EpH4) transformed with oncogenic Ras (Ep2Ras), we showed that a Ras-induced hyperactive extracellular signal-regulated kinase (ERK)/ mitogen-activated protein kinase (MAPK) signaling cooperating with TGFb is required to cause EMT in vitro and in vivo and tumor metastasis in nude mice (Oft et al, 1996;Janda et al, 2002a).…”

Section: Introductionmentioning

confidence: 99%

Raf plus TGFβ-dependent EMT is initiated by endocytosis and lysosomal degradation of E-cadherin

Janda

Nevolo

Lehmann³

et al. 2006

Oncogene

147

116

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Oncogenic Ras interferes with adhesive functions of epithelial cells, but requires tumor growth factor b (TGFb) signaling to cause epithelial-mesenchymal transition (EMT) and tumor progression in model systems. To investigate the mechanisms by which Ras and TGFb pathways cooperate in EMT induction, we introduced a tamoxifen-inducible version of Raf-1 (RafER) into fully polarized, mammary epithelial cells (EpH4). EMT characterized by loss of E-cadherin expression and upregulation of invasiveness-promoting genes was induced by TGFb plus 4-hydroxytamoxifen (4HT) activation of RafER. Downregulation of E-cadherin by RafER plus TGFb was detectable in total cell lysates after 48 h and much earlier in detergent-insoluble fractions of E-cadherin. Both pathways cooperated to strongly enhance endocytosis of E-cadherin, mainly via the clathrin-dependent route. Pulse-chase experiments showed decreased E-cadherin protein stability in cells stimulated with TGFb and 4HT and increased E-cadherin half-life in the presence of monensin. Monensin and chloroquine prevented E-cadherin degradation to different extent, but only monensin effectively blocked the loss of E-cadherin from the junctional complexes. Both lysosome inhibitors caused accumulation of E-cadherin vesicles, some of which were positive for Cathepsin D and lysosome-associated membrane protein 1 (LAMP-1). In addition, TGFb and mitogen-activated protein kinase hyperactivation synergistically induced E-cadherin ubiquitination, suggesting that the cooperation of Raf and TGFb favors lysosomal degradation of E-cadherin instead of its recycling. Our data indicate that early stages of EMT involve cooperative, post-translational downregulation of E-cadherin, whereas loss of E-cadherin via transcriptional repression is a late event in EMT.

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“…Extrinsic signals include transforming growth factor beta, hepatocyte growth factor, epidermal growth factor, and fibroblast growth factor overexpression 11, 12, 13, 14. The loss of E‐cadherin is related to a higher risk of metastasis, increased tumor grade, and lower OS 15, 16, 17, 18.…”

Section: Introductionmentioning

confidence: 99%

Epithelial–mesenchymal transition, proliferation, and angiogenesis in locally advanced cervical cancer treated with chemoradiotherapy

Rojas‐Puentes

Cardona²,

Carranza³

et al. 2016

Cancer Medicine

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We evaluated the association between epithelial–mesenchymal transition (EMT)‐derived markers and expression of proteins associated with cell proliferation and tumor growth, as well as their prognostic roles, in 61 patients (mean age 52 ± 10 years) with locally advanced cervical cancer, all of whom were treated with chemoradiation and intracavitary brachytherapy. We used immunohistochemical analysis to assess the expression of proteins targeted in our investigation. Various statistical analyses were then conducted to assess protein marker associations with survival outcomes. Forty‐six percent of the patients were positive for human papilloma virus. Median progression‐free survival (PFS) was 6.6 months (95% confidence interval [CI]: 4.0–9.1, whereas overall survival (OS) was 30.0 months (95% CI: 11–48). Multivariate analysis demonstrated that vascular endothelial growth factor (VEGF) (P = 0.002), epidermal growth factor receptor (EGFR) (P = 0.001), and TWIST2 (P = 0.001) expression levels, as well as a tumor size <6 cm (P = 0.02), influenced OS. Changes in TWIST2 levels and loss of E‐cadherin expression were correlated with VEGF and EGFR levels; furthermore, patients with high TWIST2 expression had shorter OS (P = 0.0001), as those with loss of E‐cadherin (P = 0.02). OS was even shorter when positive EGFR or VEGF expression was related with EMT markers (positive EGFR + negative E‐cadherin: median 14 months, 95% CI: 3–24; negative EGFR + positive E‐cadherin: median 31 months, 95% CI: 14–NA; P = 0.02.). The presence of EMT markers was associated with proliferative and pro‐angiogenic protein expression and influenced the prognosis of locally advanced cervical cancer.

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Diverse cellular and molecular mechanisms contribute to epithelial plasticity and metastasis

Cited by 616 publications

References 51 publications

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

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

Raf plus TGFβ-dependent EMT is initiated by endocytosis and lysosomal degradation of E-cadherin

Epithelial–mesenchymal transition, proliferation, and angiogenesis in locally advanced cervical cancer treated with chemoradiotherapy

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