Microgravity Induces Transient EMT in Human Keratinocytes by Early Down-Regulation of E-Cadherin and Cell-Adhesion Remodeling

Changes in cell–matrix and cell-to-cell adhesion patterns are dramatically fostered by the microgravity exposure of living cells. The modification of adhesion properties could promote the emergence of a migrating and invasive phenotype. We previously demonstrated that short exposure to the simulated microgravity of human keratinocytes (HaCaT) promotes an early epithelial–mesenchymal transition (EMT). Herein, we developed this investigation to verify if the cells maintain the acquired invasive phenotype after an extended period of weightlessness exposure. We also evaluated cells’ capability in recovering epithelial characteristics when seeded again into a normal gravitational field after short microgravity exposure. We evaluated the ultra-structural junctional features of HaCaT cells by Transmission Electron Microscopy and the distribution pattern of vinculin and E-cadherin by confocal microscopy, observing a rearrangement in cell–cell and cell–matrix interactions. These results are mirrored by data provided by migration and invasion biological assay. Overall, our studies demonstrate that after extended periods of microgravity, HaCaT cells recover an epithelial phenotype by re-establishing E-cadherin-based junctions and cytoskeleton remodeling, both being instrumental in promoting a mesenchymal–epithelial transition (MET). Those findings suggest that cytoskeletal changes noticed during the first weightlessness period have a transitory character, given that they are later reversed and followed by adaptive modifications through which cells miss the acquired mesenchymal phenotype.

Download Full-text

E-cadherin represses anchorage-independent growth in sarcomas through both signaling and mechanical mechanisms

10.1101/347815 ◽

2018 ◽

Author(s):

Mohit Kumar Jolly ◽

Kathryn E Ware ◽

Shengnan Xu ◽

Shivee Gilja ◽

Samantha Shetler ◽

...

Keyword(s):

Cell Adhesion ◽

Intracellular Signaling ◽

Epithelial Mesenchymal Transition ◽

Migration And Invasion ◽

Specific Cell ◽

Mesenchymal Transition ◽

Anchorage Independent Growth ◽

Sarcoma Cells ◽

E Cadherin ◽

Cell Cell

E-cadherin, an epithelial-specific cell-cell adhesion molecule, plays multiple roles in maintaining adherens junctions, regulating migration and invasion, and mediating intracellular signaling. Downregulation of E-cadherin is a hallmark of epithelial-mesenchymal transition (EMT) and correlates with poor prognosis in multiple carcinomas. Conversely, upregulation of E-cadherin is prognostic for improved survival in sarcomas. Yet, despite the prognostic benefit of E-cadherin expression in sarcoma, the mechanistic significance of E-cadherin in sarcomas remains poorly understood. Here, by combining mathematical models with wet-bench experiments, we identify the core regulatory networks mediated by E-cadherin in sarcomas, and decipher their functional consequences. Unlike in carcinomas, E-cadherin overexpression in sarcomas does not induce a mesenchymal-epithelial transition (MET). However, E-cadherin acts to reduce both anchorage-independent growth and spheroid formation of sarcoma cells. Ectopic E-cadherin expression acts to downregulate phosphorylated CREB (p-CREB) and the transcription factor, TBX2, to inhibit anchorage-independent growth. RNAi-mediated knockdown of TBX2 phenocopies the effect of E-cadherin on p-CREB levels and restores sensitivity to anchorage-independent growth in sarcoma cells. Beyond its signaling role, E-cadherin expression in sarcoma cells can also strengthen cell-cell adhesion and restricts spheroid growth through mechanical action. Together, our results demonstrate that E-cadherin inhibits sarcoma aggressiveness by preventing anchorage-independent growth.

Download Full-text

HDAC4 promotes nasopharyngeal carcinoma progression and serves as a therapeutic target

Cell Death and Disease ◽

10.1038/s41419-021-03417-0 ◽

2021 ◽

Vol 12 (2) ◽

Author(s):

Chun Cheng ◽

Jun Yang ◽

Si-Wei Li ◽

Guofu Huang ◽

Chenxi Li ◽

...

Keyword(s):

Nasopharyngeal Carcinoma ◽

Tumor Growth ◽

Therapeutic Target ◽

Histone Deacetylases ◽

Migration And Invasion ◽

Poor Overall Survival ◽

Mesenchymal Transition ◽

E Cadherin

AbstractHistone deacetylases (HDACs) are involved in tumor progression, and some have been successfully targeted for cancer therapy. The expression of histone deacetylase 4 (HDAC4), a class IIa HDAC, was upregulated in our previous microarray screen. However, the role of HDAC4 dysregulation and mechanisms underlying tumor growth and metastasis in nasopharyngeal carcinoma (NPC) remain elusive. Here, we first confirmed that the HDAC4 levels in primary and metastatic NPC tissues were significantly increased compared with those in normal nasopharyngeal epithelial tissues and found that high HDAC4 expression predicted a poor overall survival (OS) and progression-free survival (PFS). Functionally, HDAC4 accelerated cell cycle G1/S transition and induced the epithelial-to-mesenchymal transition to promote NPC cell proliferation, migration, and invasion in vitro, as well as tumor growth and lung metastasis in vivo. Intriguingly, knockdown of N-CoR abolished the effects of HDAC4 on the invasion and migration abilities of NPC cells. Mechanistically, HDAC3/4 binds to the E-cadherin promoter to repress E-cadherin transcription. We also showed that the HDAC4 inhibitor tasquinimod suppresses tumor growth in NPC. Thus, HDAC4 may be a potential diagnostic marker and therapeutic target in patients with NPC.

Download Full-text

The Impacts of Bone Marrow Mesenchymal Stem Cells (BMSCs)-Derived Periostin on Epithelial-Mesenchymal Transition (EMT) of Cervical Cancer Cells

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2022.2958 ◽

2022 ◽

Vol 12 (4) ◽

pp. 820-826

Author(s):

Chengyong Wu ◽

Weifeng Wei ◽

Jing Li ◽

Shenglin Peng

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Epithelial Mesenchymal Transition ◽

Signal Transduction Pathway ◽

Underlying Mechanism ◽

Migration And Invasion ◽

Mesenchymal Transition ◽

Essential Components ◽

Cervical Cancer Cells ◽

E Cadherin

Epithelial-mesenchymal transition (EMT) is closely related to the migrating and invading behaviors of cells. Periostin is one of the essential components in the extracellular matrix and can induce EMT of cells and their sequential metastasis. But its underlying mechanism is unclear. The Hela and BMSC cell lines were assigned into Periostin-mimic group, Periostin-Inhibitor group and Periostin-NC group followed by analysis of cell migration and invasion, expression of E-Cadherin, Vimentin, β-Catenin, Snail, MMP-2, MMP-9, PTEN, and p-PTEN. Cells in Periostin-mimic group exhibited lowest migration, least number of invaded cells, as well as lowest levels of Vimentin, β-Catenin, Snail, MMP-2, MMP-9, p-PTEN, Akt, p-Akt, p-GSK-3β, p-PDK1 and p-cRcf, along with highest levels of E-cadherin and PTEN. Moreover, cells in Periostin-NC group had intermediate levels of these above indicators, while, the Periostin-Inhibitor group exhibited the highest migration rate, the most number of invaded cells, and the highest levels of these proteins (P < 0.05). In conclusion, BMSCs-derived Periostin can influence the EMT of cervical cancer cells possibly through restraining the activity of the PI3K/AKT signal transduction pathway, indicating that Periostin might be a target of chemotherapy in clinics for the treatment of cervical cancer.

Download Full-text

SETD1A to induce epithelial-mesenchymal transition to promote invasion and metastasis through epigenetic reprogramming of snail in gastric cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.3_suppl.241 ◽

2021 ◽

Vol 39 (3_suppl) ◽

pp. 241-241

Author(s):

Jugang Wu ◽

Jiwei Yu ◽

Yan Gu

Keyword(s):

Gastric Cancer ◽

Molecular Mechanisms ◽

Epithelial Mesenchymal Transition ◽

Epigenetic Modification ◽

Epigenetic Reprogramming ◽

Migration And Invasion ◽

Mrna Levels ◽

Invasion And Metastasis ◽

Mesenchymal Transition ◽

E Cadherin

241 Background: Aberrant epigenetic modification induces oncogenes expression and promotes cancer development. The histone lysine methyltransferase SETD1A, which specifically methylates H3K4, is involved in tumor growth and metastasis, and its ectopic expression has been detected in aggressive malignancies. Our previous study had reported that SETD1A promoted gastric cancer (GC) proliferation and tumorigenesis. However, the function and molecular mechanisms of SETD1A in GC metastasis remain to be elucidated. Methods: Transwell migration and invasion assay were performed to determine GC cell migration and invasion. Lung metastasis assay was used to detect GC cell metastasis. Western Blot and Real-time qPCR were performed to measure the protein and mRNA levels, respectively. ChIP assay was performed to investigate the methylation of H3K4. The correlation between SETD1A and EMT associated key genes in GC were performed by bioinformatic analysis. Results: In this study, we found that overexpression of SETD1A promotes GC migration and invasion, whereas knockdown of SETD1A suppressed GC migration, invasion and metastasis. Furthermore, knockdown of SETD1A suppressed GC epithelial-mesenchymal transition (EMT) by increasing the expression of epithelial marker E-cadherin, and decreasing the expression of mesenchymal markers, including N-cadherin, Fibronectin and Vimentin. Mechanistically, knockdown of SETD1A reduced the EMT key transcriptional factors snail. SETD1A was recruited to the promoter of snail, where SETD1A could methylate H3K4. However, knockdown of SETD1A decreased the methylation of H3K4 on snail promoter. Rescue of snail restored SETD1A knockdown-induced GC migration and invasion inhibition. In addition, linear correlation between SETD1A and several key EMT genes, including E-cadherin, Fibronectin and snail, in GC specimens obtained from TCGA dataset. Conclusions: In summary, our data reveals that SETD1A mediated EMT process and induced metastasis through epigenetic reprogramming of snail.

Download Full-text

Disruption of Cancer Metabolic SREBP1/miR-142-5p Suppresses Epithelial–Mesenchymal Transition and Stemness in Esophageal Carcinoma

Cells ◽

10.3390/cells9010007 ◽

2019 ◽

Vol 9 (1) ◽

pp. 7 ◽

Cited By ~ 2

Author(s):

Chih-Ming Huang ◽

Chin-Sheng Huang ◽

Tung-Nien Hsu ◽

Mao-Suan Huang ◽

Iat-Hang Fong ◽

...

Keyword(s):

Esophageal Cancer ◽

Survival Rate ◽

Tumor Suppressor ◽

Poor Prognosis ◽

Epithelial Mesenchymal Transition ◽

Therapeutic Potential ◽

Regulatory Element ◽

Migration And Invasion ◽

Mesenchymal Transition ◽

E Cadherin

Elevated activity of sterol regulatory element-binding protein 1 (SREBP1) has been implicated in the tumorigenesis of different cancer types. However, the functional roles of SREBP1 in esophageal cancer are not well appreciated. Here, we aimed to investigate the therapeutic potential of SREBP1 and associated signaling in esophageal cancer. Our initial bioinformatics analyses showed that SREBP1 expression was overexpressed in esophageal tumors and correlated with a significantly lower overall survival rate in patients. Additionally, tumor suppressor miR-142-5p was predicted to target SREBP1/ZEB1 and a lower miR-142-5p was correlated with poor prognosis. We then performed in vitro experiments and showed that overexpressing SREBP1 in OE33 cell line led to increased abilities of colony formation, migration, and invasion; the opposite was observed in SREBP1-silenced OE21cells and SREBP1-silencing was accompanied by the reduced mesenchymal markers, including vimentin (Vim) and ZEB1, while E-cadherin and tumor suppressor miR-142-5p were increased. Subsequently, we first demonstrated that both SREBP1 and ZEB1 were potential targets of miR-142-5p, followed by the examination of the regulatory circuit of miR-142-5p and SREBP1/ZEB1. We observed that increased miR-142-5p level led to the reduced tumorigenic properties, such as migration and tumor sphere formation, and both observations were accompanied by the reduction of ZEB1 and SREBP1, and increase of E-cadherin. We then explored the potential therapeutic agent targeting SREBP1-associated signaling by testing fatostatin (4-hydroxytamoxifen, an active metabolite of tamoxifen). We found that fatostatin suppressed the cell viability of OE21 and OE33 cells and tumor spheres. Interestingly, fatostatin treatment reduced CD133+ population in both OE21 and OE33 cells in concert of increased miR-142-5p level. Finally, we evaluated the efficacy of fatostatin using a xenograft mouse model. Mice treated with fatostatin showed a significantly lower tumor burden and better survival rate as compared to their control counterparts. The treatment of fatostatin resulted in the reduced staining of SREBP1, ZEB1, and Vim, while E-cadherin and miR-142-5p were increased. In summary, we showed that increased SREBP1 and reduced miR-142-5p were associated with increased tumorigenic properties of esophageal cancer cells and poor prognosis. Preclinical tests showed that suppression of SREBP1 using fatostatin led to the reduced malignant phenotype of esophageal cancer via the reduction of EMT markers and increased tumor suppressor, miR-142-5p. Further investigation is warranted for the clinical use of fatostatin for the treatment of esophageal malignancy.

Download Full-text

Inhibitor of DNA-Binding Protein 4 Suppresses Cancer Metastasis through the Regulation of Epithelial Mesenchymal Transition in Lung Adenocarcinoma

Cancers ◽

10.3390/cancers11122021 ◽

2019 ◽

Vol 11 (12) ◽

pp. 2021 ◽

Cited By ~ 1

Author(s):

Chi-Chung Wang ◽

Yuan-Ling Hsu ◽

Chi-Jen Chang ◽

Chia-Jen Wang ◽

Tzu-Hung Hsiao ◽

...

Keyword(s):

Lung Cancer ◽

Dna Binding ◽

Lung Adenocarcinoma ◽

Cancer Metastasis ◽

Epithelial Mesenchymal Transition ◽

Migration And Invasion ◽

Mesenchymal Transition ◽

Inhibitor Of Dna Binding ◽

E Cadherin

Metastasis is a predominant cause of cancer death and the major challenge in treating lung adenocarcinoma (LADC). Therefore, exploring new metastasis-related genes and their action mechanisms may provide new insights for developing a new combative approach to treat lung cancer. Previously, our research team discovered that the expression of the inhibitor of DNA binding 4 (Id4) was inversely related to cell invasiveness in LADC cells by cDNA microarray screening. However, the functional role of Id4 and its mechanism of action in lung cancer metastasis remain unclear. In this study, we report that the expression of Id4 could attenuate cell migration and invasion in vitro and cancer metastasis in vivo. Detailed analyses indicated that Id4 could promote E-cadherin expression through the binding of Slug, cause the occurrence of mesenchymal-epithelial transition (MET), and inhibit cancer metastasis. Moreover, the examination of the gene expression database (GSE31210) also revealed that high-level expression of Id4/E-cadherin and low-level expression of Slug were associated with a better clinical outcome in LADC patients. In summary, Id4 may act as a metastatic suppressor, which could not only be used as an independent predictor but also serve as a potential therapeutic for LADC treatment.

Download Full-text

Atypical role of sprouty in colorectal cancer: sprouty repression inhibits epithelial–mesenchymal transition

Oncogene ◽

10.1038/onc.2015.365 ◽

2015 ◽

Vol 35 (24) ◽

pp. 3151-3162 ◽

Cited By ~ 16

Author(s):

Q Zhang ◽

T Wei ◽

K Shim ◽

K Wright ◽

K Xu ◽

...

Keyword(s):

Colorectal Cancer ◽

Colon Cancer ◽

Cell Migration ◽

Cancer Cells ◽

Epithelial Mesenchymal Transition ◽

Migration And Invasion ◽

Colon Cancer Cells ◽

Mesenchymal Transition ◽

E Cadherin

Abstract Sprouty (SPRY) appears to act as a tumor suppressor in cancer, whereas we demonstrated that SPRY2 functions as a putative oncogene in colorectal cancer (CRC) (Oncogene, 2010, 29: 5241–5253). We investigated the mechanisms by which SPRY regulates epithelial–mesenchymal transition (EMT) in CRC. SPRY1 and SPRY2 mRNA transcripts were significantly upregulated in human CRC. Suppression of SPRY2 repressed AKT2 and EMT-inducing transcription factors and significantly increased E-cadherin expression. Concurrent downregulation of SPRY1 and SPRY2 also increased E-cadherin and suppressed mesenchymal markers in colon cancer cells. An inverse expression pattern between AKT2 and E-cadherin was established in a human CRC tissue microarray. SPRY2 negatively regulated miR-194-5p that interacts with AKT2 3′ untranslated region. Mir-194 mimics increased E-cadherin expression and suppressed cancer cell migration and invasion. By confocal microscopy, we demonstrated redistribution of E-cadherin to plasma membrane in colon cancer cells transfected with miR-194. Spry1 −/− and Spry2 −/− double mutant mouse embryonic fibroblasts exhibited decreased cell migration while acquiring several epithelial markers. In CRC, SPRY drive EMT and may serve as a biomarker of poor prognosis.

Download Full-text

Transformation of cell adhesion properties by exogenously introduced E-cadherin cDNA

Nature ◽

10.1038/329341a0 ◽

1987 ◽

Vol 329 (6137) ◽

pp. 341-343 ◽

Cited By ~ 451

Author(s):

Akira Nagafuchi ◽

Yasuaki Shirayoshi ◽

Kenji Okazaki ◽

Kunio Yasuda ◽

Masatoshi Takeichi

Keyword(s):

Cell Adhesion ◽

Adhesion Properties ◽

E Cadherin

Download Full-text

Immediate and Delayed Effects of E-Cadherin Inhibition on Gene Regulation and Cell Motility in Human Epidermoid Carcinoma Cells

Molecular and Cellular Biology ◽

10.1128/mcb.25.20.9138-9150.2005 ◽

2005 ◽

Vol 25 (20) ◽

pp. 9138-9150 ◽

Cited By ~ 39

Author(s):

Henriette Andersen ◽

Jakob Mejlvang ◽

Shaukat Mahmood ◽

Irina Gromova ◽

Pavel Gromov ◽

...

Keyword(s):

Cell Adhesion ◽

Cell Motility ◽

Family Members ◽

Dominant Negative ◽

Dominant Negative Mutant ◽

Short Term ◽

Mesenchymal Transition ◽

Negative Mutant ◽

E Cadherin ◽

Cell Cell

ABSTRACT The invasion suppressor protein, E-cadherin, plays a central role in epithelial cell-cell adhesion. Loss of E-cadherin expression or function in various tumors of epithelial origin is associated with a more invasive phenotype. In this study, by expressing a dominant-negative mutant of E-cadherin (Ec1WVM) in A431 cells, we demonstrated that specific inhibition of E-cadherin-dependent cell-cell adhesion led to the genetic reprogramming of tumor cells. In particular, prolonged inhibition of cell-cell adhesion activated expression of vimentin and repressed cytokeratins, suggesting that the effects of Ec1WVM can be classified as epithelial-mesenchymal transition. Both short-term and prolonged expression of Ec1WVM resulted in morphological transformation and increased cell migration though to different extents. Short-term expression of Ec1WVM up-regulated two AP-1 family members, c-jun and fra-1, but was insufficient to induce complete mesenchymal transition. AP-1 activity induced by the short-term expression of Ec1WVM was required for transcriptional up-regulation of AP-1 family members and down-regulation of two other Ec1WVM-responsive genes, S100A4 and igfbp-3. Using a dominant-negative mutant of c-Jun (TAM67) and RNA interference-mediated silencing of c-Jun and Fra-1, we demonstrated that AP-1 was required for cell motility stimulated by the expression of Ec1WVM. In contrast, Ec1WVM-mediated changes in cell morphology were AP-1-independent. Our data suggest that mesenchymal transition induced by prolonged functional inhibition of E-cadherin is a slow and gradual process. At the initial step of this process, Ec1WVM triggers a positive autoregulatory mechanism that increases AP-1 activity. Activated AP-1 in turn contributes to Ec1WVM-mediated effects on gene expression and tumor cell motility. These data provide novel insight into the tumor suppressor function of E-cadherin.

Download Full-text

FOXM1 Drives Epithelial-Mesenchymal Transition Program Through E-Cadherin Promoter Biding Ability in Non-Small-Cell Lung Cancer Cells

10.21203/rs.3.rs-702896/v1 ◽

2021 ◽

Author(s):

Ling Wang ◽

Lin Xiaolan ◽

ZongSheng Jiang ◽

Yanzi Sun ◽

Yixuan Li ◽

...

Keyword(s):

Cell Migration ◽

Enzyme Assay ◽

Epithelial Mesenchymal Transition ◽

Small Cell ◽

Migration And Invasion ◽

Small Cell Lung ◽

Mesenchymal Transition ◽

Reporter Enzyme ◽

Tgf Β1 ◽

E Cadherin

Abstract Background: Forkhead box (FOX) gene family plays a critical role in regulating Epithelial-mesenchymal transition (EMT) program, and in which, FOXM1 can mediate multiple malignant process in many type of tumor cells. However, the modulate functions of FOXM1 on EMT in non-small-cell lung cancer (NSCLC) cells, especially the transcriptional function on E-cadherin coding gene CDH1 remains unclear. This article mainly focuses on FOXM1, exploring its mechanism in regulating EMT of NSCLC cells, and FOXM1 inhibitor thiostrepton’s effects in EMT intervention. Methods: Morphological changes of overexpressed cells were observed by HE staining. The effects of scratch test, Transwell chamber test and Western-blot analysis on cell migration and invasion ability and the expression of EMT-related markers were analyzed. Dual luciferin reporter enzyme assay and nuclear transcription factor immunoprecipitation assay (ChIP, immunofluorescence) revealed the transcriptional regulation of FOXM1 on EMT markers. MTT assay and clone formation assay were used to determine the effect of thiomycin on the viability of NSCLC cells and the ability of cell clone formation.Rusults: After overexpression of FOXM1, the cells showed intermediate epithelial-mesenchymal morphology, but not complete mesenchymal morphology, and their migration and invasion abilities were enhanced. The protein expression levels of N-cadherin,Snail1 and Vimentin were increased, while the expression levels of E-cadherin were decreased. On the contrary, knockdown of FOXM1 expression showed the opposite result. The double luciferin reporter enzyme assay showed that FOXM1 inhibited the luciferin reporter vector CDH1-2000-promoter. ChIP results confirmed that FOXM1 could bind endogenous to CDH1 gene promoter. In cells overexpressing FOXM1, knockdown of Snail further promotes FOXM1-mediated CDH1 transcription. MTT results and clone formation experiments showed that thiomycin had inhibitory effect on the proliferation of NSCLC cells. Morphological observation, cell migration assay and Transwell chamber assay showed that streptotin inhibited TGF-β1-induced enhanced cell migration and invasion. Western-blot analysis showed that thiomycin down-regulated the expression of FOXM1, N-cadherin, Snail, and Vimentin induced by TGF-β1, while blocking the expression of E-cadherin induced by TGF-β1 decreased.Conclusion: FOXM1 can directly bind to the promoter of E-cadherin encoding gene, and can indirectly inhibit E-cadherin expression by stimulating Snail. Overexpression of FOXM1 can promote EMT progression in NSCLC cells. Therefore, down-regulation of FOXM1 can inhibit this process. In addition, thiostrepton, a FOXM1 inhibitor, blocked proliferation, colony formation, and EMT progression in NSCLC cells.

Download Full-text