scholarly journals FGF signaling mediates definitive endoderm formation by regulating epithelial-to-mesenchymal transition and cell proliferation

2020 ◽  
Vol 64 (10-11-12) ◽  
pp. 471-477
Author(s):  
Shengbiao Li ◽  
Qingsong Huang ◽  
Jianwen Mao ◽  
Qiuhong LI
Keyword(s):  
Cell Proliferation ◽  
Epithelial To Mesenchymal Transition ◽  
Embryonic Stem ◽  
Selective Inhibitor ◽  
Definitive Endoderm ◽  
Fgf Signaling ◽  
Adhesion Protein ◽  
Mesenchymal Transition ◽  
Zinc Finger Transcription Factor

FGF signaling pathway is imperative for definitive endoderm (DE) differentiation from human embryonic stem cells (hESCs), which always accompanies an epithelial-to-mesenchymal transition (EMT) process. However, whether there is an association between FGF signaling and the EMT during DE formation in vitro has remained elusive. In the present study, we identify that several FGF family members were significantly activated during the differentiation of hESCs toward DE. Inhibition of FGF signaling by an efficient and selective inhibitor BGJ398 abolishes both the EMT and DE induction by blocking the activation of the zinc-finger transcription factor SNAI1 which is a direct transcriptional repressor of cell adhesion protein CDH1. In addition, cell proliferation is also severely influenced by attenuating the FGF signaling. Collectively, we propose that the FGF signaling promotes the DE formation through mediating the EMT and cell proliferation.

scholarly journals Sp1-Induced Upregulation of LBX2-AS1 Aggravates The Progression of Glioblastoma By Targeting The miR-491-5p/LIF Axis

2021 ◽  
Author(s):  
Wentao Li ◽  
Ismatullah Soufiany ◽  
Xiao Lyu ◽  
Lin Zhao ◽  
Chenfei Lu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Lines ◽  
Cancer Progression ◽  
Epithelial To Mesenchymal Transition ◽  
Luciferase Reporter ◽  
Mesenchymal Transition ◽  
Stat3 Signaling ◽  
Regulatory Effects

Abstract Background: Mounting evidences have shown the importance of lncRNAs in tumorigenesis and cancer progression. LBX2-AS1 is an oncogenic lncRNA that has been found abnormally expressed in gastric cancer and lung cancer samples. Nevertheless, the biological function of LBX2-AS1 in glioblastoma (GBM) and potential molecular mechanism are largely unclear. Methods: Relative levels of LBX2-AS1 in GBM samples and cell lines were detected by qRT-PCR and FISH. In vivo and in vitro regulatory effects of LBX2-AS1 on cell proliferation, epithelial-to-mesenchymal transition (EMT) and angiogenesis in GBM were examined through xenograft models and functional experiments, respectively. The interaction between Sp1 and LBX2-AS1 was assessed by ChIP. Through bioinformatic analyses, dual-luciferase reporter assay, RIP and Western blot, the regulation of LBX2-AS1 and miR-491-5p on the target gene leukemia Inhibitory factor (LIF) was identified. Results: LBX2-AS1 was upregulated in GBM samples and cell lines, and its transcription was promoted by binding to the transcription factor Sp1. As a lncRNA mainly distributed in the cytoplasm, LBX2-AS1 upregulated LIF, and activated the LIF/STAT3 signaling by exerting the miRNA sponge effect on miR-491-5p, thus promoting cell proliferation, EMT and angiogenesis in GBM. Besides, LBX2-AS1 was unfavorable to the progression of glioma and the survival. Conclusion: Upregulated by Sp1, LBX2-AS1 promotes the progression of GBM by targeting the miR-491-5p/LIF axis. It is suggested that LBX2-AS1 may be a novel diagnostic biomarker and therapeutic target of GBM.

scholarly journals Molecular mechanism of symmetry breaking in a 3D model of a human epiblast

2018 ◽  
Author(s):  
Mijo Simunovic ◽  
Jakob J. Metzger ◽  
Fred Etoc ◽  
Anna Yoney ◽  
Albert Ruzo ◽  
...  
Keyword(s):  
Symmetry Breaking ◽  
Axial Symmetry ◽  
3D Model ◽  
Epithelial To Mesenchymal Transition ◽  
Embryonic Stem ◽  
Primitive Streak ◽  
Molecular Evidence ◽  
Axis Formation ◽  
Mesenchymal Transition

ABSTRACTBreaking the anterior-posterior (AP) symmetry in mammals takes place at gastrulation. Much of the signaling network underlying this process has been elucidated in the mouse, however there is no direct molecular evidence of events driving axis formation in humans. Here, we use human embryonic stem cells to generate an in vitro 3D model of a human epiblast whose size, cell polarity, and gene expression are similar to a 10-day human epiblast. A defined dose of bone mor-phogenetic protein 4 (BMP4) spontaneously breaks axial symmetry, and induces markers of the primitive streak and epithelial to mesenchymal transition. By gene knockouts and live-cell imaging we show that, downstream of BMP4, WNT3 and its inhibitor DKK1 play key roles in this process. Our work demonstrates that a model human epiblast can break axial symmetry despite no asymmetry in the initial signal and in the absence of extraembryonic tissues or maternal cues. Our 3D model opens routes to capturing molecular events underlying axial symmetry breaking phenomena, which have largely been unexplored in model human systems.

scholarly journals BMP4-induced symmetry breaking in a 3D model of the human epiblast

2019 ◽  
Author(s):  
Mijo Simunovic ◽  
Ali H. Brivanlou ◽  
Eric D. Siggia
Keyword(s):  
Live Cell Imaging ◽  
3D Model ◽  
Epithelial To Mesenchymal Transition ◽  
Cell Imaging ◽  
Embryonic Stem ◽  
Primitive Streak ◽  
Mesenchymal Transition ◽  
Pluripotency Markers ◽  
Anterior Posterior

Abstract We describe the protocol of generating a 3D stem-cell-based model of the human pre-gastrulation epiblast by culturing human embryonic stem cells in a mix of hydrogel and Matrigel. Much like the epiblast of an in vitro attached day-10 human embryo, this model is an epithelial sphere with a cavity at its center, it is expressing key pluripotency markers, and it displays apico-basal polarity. The 3D colonies can further be differentiated with morphogens and in the case of intermediate concentrations of BMP4, they break the anterior-posterior symmetry characterized by an asymmetric expression of a primitive streak marker and showing signs of epithelial to mesenchymal transition. The protocol described here is suitable for immunofluorescence staining and for live-cell imaging.

scholarly journals Endoderm and Hepatic Progenitor Cells Engraft in the Quiescent Liver Concurrent with Intrinsically Activated Epithelial-to-Mesenchymal Transition

2021 ◽  
Vol 30 ◽  
pp. 096368972199378
Author(s):  
W. Samuel Fagg ◽  
Naiyou Liu ◽  
Igor Patrikeev ◽  
Omar A. Saldarriaga ◽  
Massoud Motamedi ◽  
...  
Keyword(s):  
Progenitor Cell ◽  
Iron Nanoparticles ◽  
Epithelial To Mesenchymal Transition ◽  
High Efficiency ◽  
Embryonic Stem ◽  
Mesenchymal Transition ◽  
Cell Engraftment ◽  
Resolution Imaging

Stem cell transplantation to the liver is a promising therapeutic strategy for a variety of disorders. Hepatocyte transplantation has short-term efficacy but can be problematic due to portal hypertension, inflammation, and sinusoidal thrombosis. We have previously transplanted small mouse endoderm progenitor (EP) cells to successfully reverse a murine model of hemophilia B, and labeling these cells with iron nanoparticles renders them responsive to magnetic fields, which can be used to enhance engraftment. The mechanisms mediating progenitor cell migration from the sinusoidal space to the hepatocyte compartment are unknown. Here we find human EP and hepatic progenitor (HP) cells can be produced from human embryonic stem cells with high efficiency, and they also readily uptake iron nanoparticles. This provides a simple manner through which one can readily identify transplanted cells in vivo using electron microscopy, shortly after delivery. High resolution imaging shows progenitor cell morphologies consistent with epithelial-to-mesenchymal transition (EMT) mediating invasion into the hepatic parenchyma. This occurs in as little as 3 h, which is considerably faster than observed when hepatocytes are transplanted. We confirmed activated EMT in transplanted cells in vitro, as well as in vivo 24 h after transplantation. We conclude that EMT naturally occurs concurrent with EP and HP cell engraftment, which may mediate the rate, safety, and efficacy of early cell engraftment in the undamaged quiescent liver.

scholarly journals PI3K/Akt1 signalling specifies foregut precursors by generating regionalized extra-cellular matrix

eLife ◽  
2013 ◽  
Vol 2 ◽  
Author(s):  
S Nahuel Villegas ◽  
Michaela Rothová ◽  
Martin E Barrios-Llerena ◽  
Maria Pulina ◽  
Anna-Katerina Hadjantonakis ◽  
...  
Keyword(s):  
Epithelial To Mesenchymal Transition ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Central Question ◽  
Mesenchymal Transition ◽  
Pi3k Activity ◽  
Pi3k Signalling ◽  
Low Levels

During embryonic development signalling pathways act repeatedly in different contexts to pattern the emerging germ layers. Understanding how these different responses are regulated is a central question for developmental biology. In this study, we used mouse embryonic stem cell (mESC) differentiation to uncover a new mechanism for PI3K signalling that is required for endoderm specification. We found that PI3K signalling promotes the transition from naïve endoderm precursors into committed anterior endoderm. PI3K promoted commitment via an atypical activity that delimited epithelial-to-mesenchymal transition (EMT). Akt1 transduced this activity via modifications to the extracellular matrix (ECM) and appropriate ECM could itself induce anterior endodermal identity in the absence of PI3K signalling. PI3K/Akt1-modified ECM contained low levels of Fibronectin (Fn1) and we found that Fn1 dose was key to specifying anterior endodermal identity in vivo and in vitro. Thus, localized PI3K activity affects ECM composition and ECM in turn patterns the endoderm.

Effect of pericytes on epithelial-to-mesenchymal transition in melanoma.

2012 ◽  
Vol 30 (30_suppl) ◽  
pp. 84-84 ◽  
Author(s):  
Jose Humberto Trevino-Villarreal ◽  
Rosalinda Sepulveda ◽  
Douglas A. Cotanche ◽  
Rick A. Rogers
Keyword(s):  
Cell Proliferation ◽  
Tumor Growth ◽  
Cancer Cells ◽  
Epithelial To Mesenchymal Transition ◽  
Scid Mice ◽  
Proliferation Rate ◽  
Mesenchymal Transition ◽  
Facs Sorting ◽  
E Cadherin

84 Background: Melanoma-associated stroma cells play an important role in supporting cancer cell proliferation, invasion, and dissemination. Increased invasiveness has been observed in cancer cells undergoing epithelial-to-mesenchymal transition (EMT); however, it is unknown whether EMT is facilitated by stroma cells. We showed that pericytes constitute a predominant stroma subpopulation that promotes melanoma growth by interacting with cancer cells. This study investigates the mechanism by which pericytes facilitate melanoma development. Methods: GFP+ pericytes isolated from tumors and adipose tissue were co-cultured in vitro with B16 melanoma cells at a 3:1 ratio. After 4 days, proliferation of B16 cells and pericytes was assessed. Also, co-cultures were labeled for several markers and analyzed by flow cytometry (FC). Tumorigenicity was investigated by isolating B16 cells from co-cultures by FACS sorting and then injected into SCID mice to measure tumor growth. Results: B16 cells induced pericyte differentiation into FAP+ myofibroblasts. However, no change in the pericyte proliferation rate was observed. B16 cells co-cultured with pericytes, and later separated by FACS sorting, displayed faster proliferation rates in vitro, and induced increased tumor growth rates when injected into SCID mice, compared with naive B16 cells. Pericytes increased B16 cell proliferation rate and induced a change in cell morphology, from cobblestone to fibroblast-like colonies. Interaction between B16 cells and pericytes in co-culture increased pericyte production of TGF-β and increased the expression of the stem cell markers SSEA-1, OCT3/4 and CD271 in B16 cells. In addition, cadherin switching, demonstrated by loss of E-cadherin expression and an up-regulation of the mesenchymal markers N-cadherin and vimentin was observed in B16 cells. Conclusions: Pericyte production of TGF-β promotes B16 cell development of EMT, as shown by loss of E-cadherin expression and up-regulation of the mesenchymal markers N-cadherin and vimentin, resulting in an increase in melanoma cell tumorigenicity. These results suggest that therapies targeting stromal pericytes may be a promising approach for melanoma treatment.

scholarly journals PPP3CB Inhibits Migration of G401 Cells via Regulating Epithelial-to-Mesenchymal Transition and Promotes G401 Cells Growth

2019 ◽  
Vol 20 (2) ◽  
pp. 275 ◽  
Author(s):  
Lei Chen ◽  
Qingling He ◽  
Yamin Liu ◽  
Yafei Wu ◽  
Dongsheng Ni ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Migration ◽  
Tumor Cells ◽  
Epithelial To Mesenchymal Transition ◽  
Phosphatase Domain ◽  
Mesenchymal Transition ◽  
In Vivo Experiments ◽  
E Cadherin

PPP3CB belongs to the phosphoprotein phosphatases (PPPs) group. Although the majority of the PPP family play important roles in the epithelial-to-mesenchymal transition (EMT) of tumor cells, little is known about the function of PPP3CB in the EMT process. Here, we found PPP3CB had high expression in kidney mesenchymal-like cells compared with kidney epithelial-like cells. Knock-down of PPP3CB downregulated epithelial marker E-cadherin and upregulated mesenchymal marker Vimentin, promoting the transition of cell states from epithelial to mesenchymal and reorganizing the actin cytoskeleton which contributed to cell migration. Conversely, overexpression of PPP3CB reversed EMT and inhibited migration of tumor cells. Besides, in vitro and in vivo experiments indicated that the loss of PPP3CB suppressed the tumor growth. However, the deletion of the phosphatase domain of PPP3CB showed no effect on the expression of E-cadherin, migration, and G401 cell proliferation. Together, we demonstrate that PPP3CB inhibits G401 cell migration through regulating EMT and promotes cell proliferation, which are both associated with the phosphatase activity of PPP3CB.

Recombinant mouse SPARC promotes parietal endoderm differentiation and cardiomyogenesis in embryoid bodies

2008 ◽  
Vol 86 (6) ◽  
pp. 487-499 ◽  
Author(s):  
Christopher Hrabchak ◽  
Maurice Ringuette ◽  
Kimberly Woodhouse
Keyword(s):  
Epithelial To Mesenchymal Transition ◽  
Embryonic Stem ◽  
Recombinant Baculovirus ◽  
Neuronal Cell ◽  
Embryoid Bodies ◽  
Marker Genes ◽  
Mesenchymal Transition ◽  
Enhanced Expression ◽  
Parietal Endoderm

In the absence of leukemia inhibitory factor, murine embryonic stem cells cultured in vitro spontaneously aggregate to from three-dimensional embryoid bodies that differentiate to produce hematopoietic, endothelial, muscle, and neuronal cell lineages in a manner recapitulating the events of early embryogenesis. Cardiomyogenesis in embryoid bodies was recently demonstrated to be promoted by PYS-2-derived native SPARC (secreted protein, acidic and rich in cysteine), whose expression is upregulated in parietal endoderm at the onset of the epithelial to mesenchymal transition. Here, we confirm the stimulatory effects of mouse SPARC on cardiomyogenesis using a recombinant baculovirus-produced protein (rmSPARC). Embryoid bodies cultured in the presence of glycosylated rmSPARC, or an unglycosylated peptide spanning the C-terminal EF-hand domain, developed greater numbers of beating cardiomyocytes than did time-matched controls, with enhanced expression of cardiac marker genes including Nkx2.5, Troponin, BMP-2, and MHCα. Histochemical analysis revealed an expansion of the peripheral endoderm, with thicker layers of extracellular matrix (ECM) material observed atop underlying cells. Embryoid bodies treated with SPARC also displayed increased adherence to polystyrene culture dishes, with enhanced expression of ECM mRNAs including collagen IVα3, collagen IVα5, and laminin α1. These results indicate that, in addition to the promotion of cardiomyogenesis, SPARC may also help regulate the molecular composition and organization of ECM secreted by the mesenchymal parietal endoderm.

scholarly journals NRBP2 Functions as a Tumor Suppressor and Inhibits Epithelial-to-Mesenchymal Transition in Breast Cancer

2021 ◽  
Vol 11 ◽  
Author(s):  
Zhiyu Li ◽  
Bingxiong Liu ◽  
Chenyuan Li ◽  
Si Sun ◽  
Hanpu Zhang ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Proliferation ◽  
Epithelial To Mesenchymal Transition ◽  
Epithelial Mesenchymal Transition ◽  
Mammalian Target Of Rapamycin ◽  
Tumor Model ◽  
Adenosine Monophosphate ◽  
Prognostic Indicator ◽  
Mesenchymal Transition

Nuclear Receptor Binding Protein 2 (NRBP2), one of the pseudokinases discovered during a screen of neural differentiation genes, inhibits tumor progression in medulloblastoma and hepatocellular carcinoma. However, the role and the mechanism of NRBP2 in the regulation of the progression of breast cancer (BC) have not been reported. In our study, NRBP2 was downregulated in human BC tissues compared with the corresponding normal tissues. Moreover, bioinformatics and cellular experiments illustrated that a lower level of NRBP2 contributed to a poor prognosis for patients with BC. In addition, we characterized the NRBP2-overexpressing BC cells and found that NRBP2 overexpression dramatically suppressed cell proliferation and invasion and inhibited the epithelial-mesenchymal transition (EMT) in cells in vitro, whereas knockdown of NRBP2 reversed these effects. Furthermore, overexpression of NRBP2 in the orthotopic breast tumor model significantly reduced lung metastatic nodules in nude mice. Mechanistically, NRBP2 regulated the activation of the 5′-adenosine monophosphate (AMP)-activated protein kinase/ mammalian target of rapamycin (AMPK/mTOR) signaling pathway. Moreover, the inhibition of cell proliferation, invasion and the EMT by NRBP2 overexpression was partially rescued after treatment with an AMPK inhibitor. Conversely, mTOR-specific inhibitors eliminated the effects of NRBP2 knockdown on increasing cell proliferation, invasion and the EMT, which suggested the anti-tumor effect of NRBP2, which may be partially related to the regulation of the AMPK/mTOR pathway. Taken together, NRBP2, a novel and effective prognostic indicator, inhibited the progression of BC and may become a potential therapeutic target for BC.

scholarly journals Cholest-4,6-Dien-3-One Promote Epithelial-To-Mesenchymal Transition (EMT) in Biliary Tree Stem/Progenitor Cell Cultures In Vitro

Cells ◽  
2019 ◽  
Vol 8 (11) ◽  
pp. 1443
Author(s):  
Lorenzo Nevi ◽  
Daniele Costantini ◽  
Samira Safarikia ◽  
Sabina Di Matteo ◽  
Fabio Melandro ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
Epithelial To Mesenchymal Transition ◽  
Biliary Tree ◽  
Telomerase Activity ◽  
Mesenchymal Transition ◽  
Peribiliary Glands ◽  
Emt Markers ◽  
Tree Stem

Human biliary tree stem/progenitor cells (hBTSCs), reside in peribiliary glands, are mainly stimulated by primary sclerosing cholangitis (PSC) and cholangiocarcinoma. In these pathologies, hBTSCs displayed epithelial-to-mesenchymal transition (EMT), senescence characteristics, and impaired differentiation. Here, we investigated the effects of cholest-4,6-dien-3-one, an oxysterol involved in cholangiopathies, on hBTSCs biology. hBTSCs were isolated from donor organs, cultured in self-renewal control conditions, differentiated in mature cholangiocytes by specifically tailored medium, or exposed for 10 days to concentration of cholest-4,6-dien-3-one (0.14 mM). Viability, proliferation, senescence, EMT genes expression, telomerase activity, interleukin 6 (IL6) secretion, differentiation capacity, and HDAC6 gene expression were analyzed. Although the effect of cholest-4,6-dien-3-one was not detected on hBTSCs viability, we found a significant increase in cell proliferation, senescence, and IL6 secretion. Interestingly, cholest-4.6-dien-3-one impaired differentiation in mature cholangiocytes and, simultaneously, induced the EMT markers, significantly reduced the telomerase activity, and induced HDAC6 gene expression. Moreover, cholest-4,6-dien-3-one enhanced bone morphogenic protein 4 (Bmp-4) and sonic hedgehog (Shh) pathways in hBTSCs. The same pathways activated by human recombinant proteins induced the expression of EMT markers in hBTSCs. In conclusion, we demonstrated that chronic exposition of cholest-4,6-dien-3-one induced cell proliferation, EMT markers, and senescence in hBTSC, and also impaired the differentiation in mature cholangiocytes.

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