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E-cadherin loss altered cytoskeletal organization in MCF10A CDH1-/- cells. a) Loss of E-cadherin altered tubulin cytoskeletal arrangement. On the apical surface of MCF10A cells, the microtubules showed radial pattern of organization (indicated by white arrows) with the minus end densely anchored in the centre and the plus end extending towards the cell cortex. However, in MCF10A CDH1 -/- cells had gross defect in the radial pattern of organization and often oriented parallel to the cortex (indicated by white arrows). At the basal surface, the microtubules form a meshwork like structure with no striking difference observed between the two cell lines. b) Loss of E-cadherin altered actin cytoskeletal arrangement. On the apical surface of MCF10A cells, actin forms cross-linking filamentous meshwork while basally it organizes itself into stress fibres like structure. Overall apical actin meshwork looks similar in both MCF10A isogenic cells but basally there are more and thicker stress fibres in MCF10A CDH1-/- cells (indicated by white arrows).
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E-cadherin is an adherens junction protein that forms homophilic intercellular contacts in epithelial cells while also interacting with the intracellular cytoskeletal networks. It has roles including establishment and maintenance of cell polarity, differentiation, migration and signalling in cell proliferation pathways. Its downregulation is common...
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Chronic inflammation orchestrates the tumor microenvironment and is strongly associated with cancer. Tumor necrosis factor- α (TNF α ) is involved in tumor invasion and metastasis by inducing epithelial to mesenchymal transition (EMT). This process is defined by the loss of epithelial characteristics and gain of mesenchymal traits. The mechanisms o...
Citations
... The epithelial-mesenchymal transition (EMT) is a biological process in which epithelial cells lose their epithelial features and undergo various metabolic changes to produce a mesenchymal phenotype [14]. Functional annotation and enrichment analysis revealed that MGT cell lines have a high expression of EMT-related It is known that during the EMT process, expression of Vimentin and α-SMA is dramatically increased, while E-Cadherin an epithelial marker is decreased [15]. Furthermore, the MGT cell lines lack E-cadherin expression while expressing high levels of vimentin and α-SMA. ...
Canine mammary gland tumors (MGT) have a poor prognosis in intact female canines, posing a clinical challenge. This study aimed to establish novel canine mammary cancer cell lines from primary tumors and characterize their cellular and molecular features to find potential therapeutic drugs. The MGT cell lines demonstrated rapid cell proliferation and colony formation in an anchorage-independent manner. Vimentin and α-SMA levels were significantly elevated in MGT cell lines compared to normal canine kidney (MDCK) cells, while CDH1 expression was either significantly lower or not detected at all, based on quantitative real-time PCR (qRT-PCR) analysis. Functional annotation and enrichment analysis revealed that epithelial-mesenchymal transition (EMT) phenotypes and tumor-associated pathways, particularly the PI3K/Akt signaling pathway, were upregulated in MGT cells. BYL719 (Alpelisib), a PI3K inhibitor, was also examined for cytotoxicity on the MGT cell lines. The results show that BYL719 can significantly inhibit the proliferation of MGT cell lines in vitro. Overall, our findings suggest that the MGT cell lines may be valuable for future studies on the development, progression, metastasis, and management of tumors.
... Similarly, the loss of epithelial state and gain of mesenchymal state during carcinogenesis might be also a secondary effect caused by signaling cascades driven initially by different factors, e.g., cancer-driving mutations in KRAS, TP53, etc. E-cadherin is a key adhesion molecule and its loss is considered as the hallmark of EMT (Sun and Ma, 2024). It is funny that E-cadherin loss does not cause an EMT effect (Chen et al., 2014). E-cadherin knockout causes defects in embryos and organs, and promotes tumorigenesis. ...
Epithelial–mesenchymal transition (EMT) is defined as a cellular process during which epithelial cells acquire mesenchymal phenotypes and behavior following the downregulation of epithelial features. EMT and its reversed process, the mesenchymal-epithelial transition (MET), and the special form of EMT, the endothelial-mesenchymal transition (EndMT), have been thought as a universal dogma controlling developmental and pathological processes. More than half a century of EMT study has made it a large research field and a mainstream concept. However, discrepancies and disputes over EMT and EMT research have also grown over time. Particularly, neither the epithelial and mesenchymal states/properties nor their regulatory networks nor specific markers have been defined, rendering the EMT, MET and EndMT concepts groundless. Moreover, EMT and MET effects should not be a cause, but rather a consequence of or an accompanying effect during developmental and pathological processes. EMT and MET represented by the change in cell shapes or adhesiveness or symbolized by EMT factors are biased interpretation of the overall change in cellular property and regulatory networks during development and cancer progression. The true meaning of EMT effects in some developmental and pathological processes, such as fibrosis, needs re-evaluation. But the core EMT factors are actually a few components of the regulatory networks of neural stemness, which determines tumorigenicity and pluripotency. The EMT effects in cancer progression and neural crest formation are wrong attribution of the role of neural stemness during cancer progression and the cell-intrinsic property of neural crest cells to the unknown mesenchymal state. It is time to reassess the significance of EMT and its related concepts in scientific research.
... Despite this, treatment with the CB1 agonists did not increase E-cadherin expression, indicating that these synthetic cannabinoids may not have the tumor suppression function that was once thought. While the majority of research indicates that a decline in E-cadherin expression is a characteristic feature of EMT, the recent literature contends that the absence of E-cadherin is neither causal nor necessary for EMT [63,64]. As such, the study findings are supported by the literature, providing further insight into the specific effects of CB1 agonists on EMT marker expression. ...
Understanding the role of biased G protein-coupled receptor (GPCR) agonism in receptor signaling may provide novel insights into the opposing effects mediated by cannabinoids, particularly in cancer and cancer metastasis. GPCRs can have more than one active state, a phenomenon called either ‘biased agonism’, ‘functional selectivity’, or ‘ligand-directed signaling’. However, there are increasing arrays of cannabinoid allosteric ligands with different degrees of modulation, called ‘biased modulation’, that can vary dramatically in a probe- and pathway-specific manner, not from simple differences in orthosteric ligand efficacy or stimulus-response coupling. Here, emerging evidence proposes the involvement of CB1 GPCRs in a novel biased GPCR signaling paradigm involving the crosstalk between neuraminidase-1 (Neu-1) and matrix metalloproteinase-9 (MMP-9) in the activation of glycosylated receptors through the modification of the receptor glycosylation state. The study findings highlighted the role of CB1 agonists AM-404, Aravnil, and Olvanil in significantly inducing Neu-1 sialidase activity in a dose-dependent fashion in RAW-Blue, PANC-1, and SW-620 cells. This approach was further substantiated by findings that the neuromedin B receptor inhibitor, BIM-23127, MMP-9 inhibitor, MMP9i, and Neu-1 inhibitor, oseltamivir phosphate, could specifically block CB1 agonist-induced Neu-1 sialidase activity. Additionally, we found that CB1 receptors exist in a multimeric receptor complex with Neu-1 in naïve, unstimulated RAW-Blue, PANC-1, and SW-620 cells. This complex implies a molecular link that regulates the interaction and signaling mechanism among these molecules present on the cell surface. Moreover, the study results demonstrate that CB1 agonists induce NFκB-dependent secretory alkaline phosphatase (SEAP) activity in influencing the expression of epithelial–mesenchymal markers, E-cadherin, and vimentin in SW-620 cells, albeit the impact on E-cadherin expression is less pronounced compared to vimentin. In essence, this innovative research begins to elucidate an entirely new molecular mechanism involving a GPCR signaling paradigm in which cannabinoids, as epigenetic stimuli, may traverse to influence gene expression and contribute to cancer and cancer metastasis.
... Upregulation of the SOX2 expression in AGS cells might also result from the hyperactivation of the JAK-STAT and PI3K signaling pathways [36][37][38]. Treatment with OLE resulted in a significant decrease in the migratory capacity of AGS cells (Fig. 7), which implies a decrease in the adhesive properties of these cells [55]. Therefore, the ability of OLE to reverse EMT and to decrease the migratory capacity of AGS cells might be due to the interference of this natural compound with the CSC phenotype in GC. ...
... Given that activation of the apoptotic pathways is one of the common cellular responses to the DNA-damaging factors (including chemo-and radiotherapy), EMT might interfere with the sensitivity of cancer cells and reduce their apoptotic response to chemotherapeutic agents [57]. The pro-apoptotic activity of OLE has been demonstrated in various cancer cells, including melanoma, glioblastoma, hepatocellular carcinoma, and breast, colon, and cervical cancers [19,[54][55][56][57][58][59][60][61][62][63]. These studies have demonstrated that the pro-apoptotic activity of OLE is related to different molecular mechanisms, such as interference with the pluripotency of CSCs, functional impairment of the mitochondria, suppression of DNA repair, development of senescence-associated secretion phenotype, and inhibition of angiogenesis and invasion [18,19,26,58,60,[62][63][64][65][66]. ...
The overall survival of patients with the advanced and recurrent gastric cancer (GC) remains unfavorable. In particular, this is due to cancer spreading and resistance to chemotherapy associated with the epithelial-mesenchymal transition (EMT) of tumor cells. EMT can be identified by the transcriptome profiling of GC for EMT markers. Indeed, analysis of the TCGA and GTEx databases (n = 408) and a cohort of GC patients (n = 43) revealed that expression of the CDH2 gene was significantly decreased in the tumors vs. non-tumor tissues and correlated with the overall survival of GC patients. Expression of the EMT-promoting transcription factors SNAIL and ZEB1 was significantly increased in GC. These data suggest that targeting the EMT might be an attractive therapeutic approach for patients with GC. Previously, we demonstrated a potent anti-cancer activity of the olive leaf extract (OLE). However, its effect on the EMT regulation in GC remained unknown. Here, we showed that OLE efficiently potentiated the inhibitory effect of the chemotherapeutic agents 5-fluorouracil (5-FU) and cisplatin (Cis) on the EMT and their pro-apoptotic activity, as was demonstrated by changes in the expression of the EMT markers (E- and N-cadherins, vimentin, claudin-1) in GC cells treated with the aforementioned chemotherapeutic agents in the presence of OLE. Thus, culturing GC cells with 5-FU + OLE or Cis + OLE attenuated the invasive properties of cancer cells. Importantly, upregulation of expression of the apoptotic markers (PARP cleaved form) and increase in the number of cells undergoing apoptosis (annexin V-positive) were observed for GC cells treated with a combination of OLE and 5-FU or Cis. Collectively, our data illustrate that OLE efficiently interferes with the EMT in GC cells and potentiates the pro-apoptotic activity of certain chemotherapeutic agents used for GC therapy.
... Deletion and dysregulation of E-cadherin are correlated with alteration of intercellular junction, and subsequently, acquire a more mobile and invasive mesenchymal-like phenotype, this allows tumor invasion and metastasis [82,83]. In view of these facts, previously E-cadherin was considerate a cause of EMT [73], however, the abatement of E-cadherin is more a consequence of EMT than a cause [84,85]. ...
The cancer continues to be a leading cause of mortality in the world, the increment number of the cases has been correlated with the mechanisms that the tumoral cells have for growth and metastasis, there are some processes for the survival and resistance of cancer cells including epithelial-mesenchymal transition, metastasis, drug resistance and angiogenesis. The EMT is a biological process that allows cells of epithelial lineage undergo to a conversion to mesenchymal cell phenotype resulting in cells in intermediate states that have both, epithelial and mesenchymal characteristics. The ETM process induced pluripotent stem cell reprogramming and confers to the cancer cells tumor-initiating and metastatic potential. The development of metastatic process involves some proteins like E-cadherin, HLA-E, SPARC, TNS-4 and FN-1, each protein has an important role as check point of the metastasis and allow immune evasion and grows the seconds tumor.
... Deletion and dysregulation of E-cadherin are correlated with alteration of intercellular junction, and subsequently, acquire a more mobile and invasive mesenchymal-like phenotype, this allows tumor invasion and metastasis [82,83]. In view of these facts, previously E-cadherin was considerate a cause of EMT [73], however, the abatement of E-cadherin is more a consequence of EMT than a cause [84,85]. ...
The cancer continues to be a leading cause of mortality in the world, the increment number of the cases has been correlated with the mechanisms that the tumoral cells have for growth and metastasis, there are some processes for the survival and resistance of cancer cells including epithelial-mesenchymal transition, metastasis, drug resistance and angiogenesis.
... Red-green scale represents down-to upregulated genes and blue represents normal expressed genes ◂ perturbations induced by the CDH1-TANGO6 deletion are greater than those induced by the CDH1 deletion alone in these pathways. Although it is widely known that E-cadherin deficiency leads to weaker adhesion and abnormal microtubule organization [50,51], which may be involved in migration upon malignant transformation [52], our data specifically link homophilic cell adhesion to the longer CDH1-TANGO6 deletion. Indeed, this clone presents upregulation of the pathway 'homophilic cell adhesion via plasma membrane adhesion molecules', which may reflect a yet to prove adhesion-compensatory mechanism, promoted by upregulation of protocadherins in the absence of E-cadherin, seen in our data. ...
Background:
Germline CDH1 pathogenic or likely pathogenic variants cause hereditary diffuse gastric cancer (HDGC). Once a genetic cause is identified, stomachs' and breasts' surveillance and/or prophylactic surgery is offered to asymptomatic CDH1 carriers, which is life-saving. Herein, we characterized an inherited mechanism responsible for extremely early-onset gastric cancer and atypical HDGC high penetrance.
Methods:
Whole-exome sequencing (WES) re-analysis was performed in an unsolved HDGC family. Accessible chromatin and CDH1 promoter interactors were evaluated in normal stomach by ATAC-seq and 4C-seq, and functional analysis was performed using CRISPR-Cas9, RNA-seq and pathway analysis.
Results:
We identified a germline heterozygous 23 Kb CDH1-TANGO6 deletion in a family with eight diffuse gastric cancers, six before age 30. Atypical HDGC high penetrance and young cancer-onset argued towards a role for the deleted region downstream of CDH1, which we proved to present accessible chromatin, and CDH1 promoter interactors in normal stomach. CRISPR-Cas9 edited cells mimicking the CDH1-TANGO6 deletion display the strongest CDH1 mRNA downregulation, more impacted adhesion-associated, type-I interferon immune-associated and oncogenic signalling pathways, compared to wild-type or CDH1-deleted cells. This finding solved an 18-year family odyssey and engaged carrier family members in a cancer prevention pathway of care.
Conclusion:
In this work, we demonstrated that regulatory elements lying down-stream of CDH1 are part of a chromatin network that control CDH1 expression and influence cell transcriptome and associated signalling pathways, likely explaining high disease penetrance and very young cancer-onset. This study highlights the importance of incorporating scientific-technological updates and clinical guidelines in routine diagnosis, given their impact in timely genetic diagnosis and disease prevention.
... However, the high expression of E-cad in the primary compared to the metastatic tumours contradicts other previous studies, which reported significantly increased E-cad expression in metastatic breast cancer in the bone compared to primary tumours [54], and an increased expression of E-cad in metastatic lung nodules compared to primaries [55]. In that setting, even though a loss of E-cad is the first step towards achieving EMT, downregulation of E-cad expression in many circumstances is inadequate to initiate EMT in tumour cells [56]. These inconsistencies may exist due to epithelialmesenchymal plasticity (EMP) in cancer cells [57], by which the tumour cells have the flexibility in moulding towards either the epithelial or mesenchymal or a hybrid state, depending on the stimulus they receive from the tumour microenvironment [58]. ...
The process of epithelial-mesenchymal transition (EMT) involves the phenotypic transformation of cells from epithelial to mesenchymal status. The cells exhibiting EMT contain features of cancer stem cells (CSC), and the dual processes are responsible for progressive cancers. Activation of hypoxia-inducible factors (HIF) is fundamental to the pathogenesis of clear cell renal cell carcinoma (ccRCC), and their role in promoting EMT and CSCs is crucial for ccRCC tumour cell survival, disease progression, and metastatic spread. In this study, we explored the status of HIF genes and their downstream targets, EMT and CSC markers, by immunohistochemistry on in-house accrued ccRCC biopsies and adjacent non-tumorous tissues from patients undergoing partial or radical nephrectomy. In combination, we comprehensively analysed the expression of HIF genes and its downstream EMT and CSC-associated targets relevant to ccRCC by using publicly available datasets, the cancer genome atlas (TCGA) and the clinical proteome tumour analysis consortium (CPTAC). The aim was to search for novel biological prognostic markers that can stratify high-risk patients likely to experience metastatic disease. Using the above two approaches, we report the development of novel gene signatures that may help to identify patients at a high risk of developing metastatic and progressive disease.
... adhesion are the key characteristics in the EM transition process [47][48][49], suggesting that the 50 alternative splicing events we got is crucial for the regulation of the EM transition process, and thus also proves the reliability of our calculating method. The above evidence shows that our method effectively selected alternative splicing events and RNA-binding proteins related to EM transition. ...
During breast cancer metastasis, the developmental process epithelial-mesenchymal (EM) transition is abnormally activated. Transcriptional regulatory networks controlling EM transition are well-studied; however, alternative RNA splicing also plays a critical regulatory role during this process. Alternative splicing was proved to control the EM transition process, and RNA-binding proteins were determined to regulate alternative splicing. A comprehensive understanding of alternative splicing and the RNA-binding proteins that regulate it during EM transition and their dynamic impact on breast cancer remains largely unknown. To accurately study the dynamic regulatory relationships, time-series data of the EM transition process are essential. However, only cross-sectional data of epithelial and mesenchymal specimens are available. Therefore, we developed a pseudotemporal causality-based Bayesian (PCB) approach to infer the dynamic regulatory relationships between alternative splicing events and RNA-binding proteins. Our study sheds light on facilitating the regulatory network-based approach to identify key RNA-binding proteins or target alternative splicing events for the diagnosis or treatment of cancers. The data and code for PCB are available at: http://hkumath.hku.hk/~wkc/PCB(data+code).zip.
... Indeed, a change in the E-cadherin levels is one of the hallmarks of an epithelial-mesenchymal transition, which is a crucial program in the regulation of cell motility and invasion. E-cadherin loss promotes a reduction in cell-substrate adhesion and causes a significant disruption to the normal organization of the microtubule and actin cytoskeletons [76]. ...
There is growing concern about the consumption of synthetic cannabinoids (SCs), one of the largest groups of new psychoactive substances, its consequence on human health (general population and workers), and the continuous placing of new SCs on the market. Although drug-induced alterations in neuronal function remain an essential component for theories of drug addiction, accumulating evidence indicates the important role of activated astrocytes, whose essential and pleiotropic role in brain physiology and pathology is well recognized. The study aims to clarify the mechanisms of neurotoxicity induced by one of the most potent SCs, named MAM-2201 (a naphthoyl-indole derivative), by applying a novel three-dimensional (3D) cell culture model, mimicking the physiological and biochemical properties of brain tissues better than traditional two-dimensional in vitro systems. Specifically, human astrocyte spheroids, generated from the D384 astrocyte cell line, were treated with different MAM-2201 concentrations (1–30 µM) and exposure times (24–48 h). MAM-2201 affected, in a concentration- and time-dependent manner, the cell growth and viability, size and morphological structure, E-cadherin and extracellular matrix, CB1-receptors, glial fibrillary acidic protein, and caspase-3/7 activity. The findings demonstrate MAM-2201-induced cytotoxicity to astrocyte spheroids, and support the use of this human 3D cell-based model as species-specific in vitro tool suitable for the evaluation of neurotoxicity induced by other SCs.
