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This study explores the interaction between immune and cancer cells in the tumor microenvironment (TME) of cervical carcinoma (CC), with emphasis on tumor-associated macrophages (M2-TAMs) and the STAT3-NF-κB signaling pathway. It investigates how Treg cell polymorphisms and TAM infiltration through these pathways influence overall survival (OS) in CC patients. This prospective study follows 100 CC patients from 2018 to 2023 using qRT-PCR and immunohistochemistry on tumor samples, and flow cytometry on blood samples to evaluate immunosuppressive cytokines and Treg cell polymorphisms. High stromal CD163+204+ TAM density, mediated by STAT3/NF-κB, correlates with biomarkers such as Ki-67, VEGFα, and FOXP3 (p < 0.001). XPO5 expression is associated with increased STAT3, SNAIL, and HPV 16/18 levels. FOXP3 T allele deletion and HLA-G polymorphism in the blood of patients correlate with higher STAT3 tumor expression and elevated IL-4 and IL-17 blood cytokines. The CXCL12-CXCR4 axis shows a strong association with STAT3, SNAIL in TME and blood cytokines, including IL-6 and IL-12. Elevated CXCL12, CXCR4, and SNAIL expression in TME significantly increases mortality risk. These findings underscore the role of M2TAM infiltration and immune modulation in tumor progression and clinical outcomes in CC. Show less

This research focuses on molecular screening of mRNA by targeting EMT regulator genes in the TGF-β/SMAD pathway to determine the difference in EMT mechanisms between non-metastatic and metastatic primary tumor cells. The method uses Real time/quantitative Polymerase Chain Reaction (RT-qPCR) to measure the expression levels of target genes in colon tissue samples from non-metastatic and metastatic patient groups. Differences in target gene expression between the two groups were analyzed using t-tests. The results of this study show significance differences in the expression of EMT-inducing genes on the TGF-β/Smad pathway between non-metastatic colorectal cancer groups and metastases. TGF-β1 (p-value : 0.041), Smad2 (p-value : 0.020), Snail1 (p-value : 0.028), Twist1 (p-value : 0.036), and ZEB1 (p-value : 0.045) gene expression was higher in the metastatic tumor group. In contrast to these genes, the expression of the Smad4 (p-value : 0.022), E-cadherin (p-value : 0.036), and vimentin (p-value : 0.048) genes was lower in the metastatic tumor group. The observed alterations in gene expression related to EMT within the TGF-β/Smad pathway in metastatic colorectal cancer are likely associated with the partial processes of EMT and MET. These alterations may contribute to further metastatic potential and increase the malignancy of the cancer. Show less

Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by a tendency to recur and a poor prognosis. Epithelial mesenchymal transition (EMT) and proliferation of nasal epithelial cells (NECs) play an important role in CRSwNP development. Secretogranin II (SCG2) is reported to be an EMT-related gene, but its role in CRSwNP has not been reported. In this study, human NECs were cultured in an air-liquid interface culture system and stimulated with IL-13 to maintain or promote the CRSwNP state. EMT-associated protein expression levels were examined by real-time quantitative PCR and Western blot. Dual luciferase, chromatin immunoprecipitation, and co-immunoprecipitation experiments were used to validate the regulatory relationship between SP1, SCG2, and ubiquitin-1 (UBQLN1). The nuclear translocation of Snail was examined by immunofluorescence assay. The results showed that the expression levels of SP1, SCG2, and UBQLN1 were all up-regulated in CRSwNP tissues. SCG2 knockdown inhibited EMT and proliferation of human NECs. Mechanistically, SP1 promoted the proliferation and EMT of human NECs by transcriptionally increasing SCG2 expression. SCG2 activated the AKT serine/threonine kinase (AKT)/glycogen synthase kinase-3 beta (GSK-3β)/Snail family transcriptional repressor 1 (Snail) pathway and promoted Snail nuclear translocation via UBQLN1. In short, SCG2, which is transcriptionally up-regulated by SP1, promotes the proliferation and EMT of human NECs by activating the AKT/GSK-3β/Snail pathway through binding to UBQLN1. Show less

Cervical Cancer is the second most common gynecological malignancy affecting a large group of women worldwide. The molecular mechanism of cervical cancer progression is still not very clear. As a result, diagnosis of cervical cancer occurs at a very advanced stage when the disease has spread to its malignant stage, causing death in the majority of women. EMT is a major culprit associated with the malignant transformation of tumor cells during cancer progression and metastasis. Hence, identification of new biomarkers to detect cervical cancer at an early stage is essential to minimize incidence and mortality. The present study aims to identify Common Differentially Expressed Genes (DEGs) and early biomarkers associated with EMT in cervical cancer. The Datasets were downloaded from the Gene Expression Omnibus (GEO) database, with Accession numbers GSE26511, GSE67522, and GSE9750. Then, the Gene Ontology (GO), KEGG pathway enrichment analysis, and protein-protein interactions (PPI) were done. Further hub genes were identified by molecular interaction networks using Cytoscape from the constructed network of DEGs. Afterwards, survival analysis was performed to assess the prognostic significance of eight hub genes associated with EMT in cervical cancer. A total of 11,339 overlapping DEGs were identified from all three datasets, among all the total 61 DEGS, and 8 hub genes were linked to the EMT pathway. Our study suggests that these eight hub genes, CDH1, CDH2, MMP2, CD44, FN1, FGF2, SNAI1, and SNAI2, may be critically associated with EMT progression. Among the eight identified EMT hub genes, CDH2 (N-cadherin) demonstrated a significant association with overall survival, while FN1 (fibronectin) was notably linked to disease-free survival, underscoring their prognostic value in cervical cancer. Based on these findings, our study suggests that CDH1, CDH2, MMP2, CD44, FN1, FGF2, SNAI1, and SNAI2 hold potential diagnostic and prognostic significance in the progression of cervical cancer. Show less

The coordination between epithelial progenitors and their mesenchymal niche is critical for organogenesis and repair, yet the mechanisms governing their competitive interactions remain unclear. Here, we reveal a paradigm of tissue-scale fitness sensing in the lung, where mesenchymal Yap levels antagonize epithelial Yap levels to dictate epithelial stem cell fate. We show that reduced fitness in alveolar fibroblasts (AF1s) via Yap/Taz or Myc deletion leads to their apoptotic elimination and a collapse of the alveolar stem cell niche. This niche collapse triggers a pathological competitive response from the epithelium, which undergoes aberrant bronchiolization that phenocopies human pulmonary fibrosis. Mechanistically, we uncovered a molecular switch that controls mesenchymal fate. During development and fibrosis resolution, Snail1/2 sequesters Yap/Taz to drive an adipogenic program, generating niche-supportive AF1s. Conversely, Yap/Taz-TEAD-Myc binding instructs a myogenic, pro-fibrotic program. Our findings demonstrate that inter-tissue cell competition, governed by a Snail/Yap rheostat, orchestrates lung architecture and provides a framework for targeting the mesenchymal niche to treat fibrotic disease. Show less

During obesity, the excessive accumulation of fat in tissue promotes dysregulated hormonal and cytokine homeostasis that triggers chronic inflammation, which is, in part, associated with an increased incidence of some cancers. This protumoral inflammatory environment is further exacerbated through the secretome of mature adipocytes, which promotes tumor angiogenesis. Emerging studies suggest that human adipocyte-derived mesenchymal stromal/stem cells (ADMSCs) may contribute to a complementary process supporting local angiogenesis termed vasculogenic mimicry (VM). The molecular mechanisms linking ADMSCs to VM and inflammation remain poorly understood. ADMSC 3D capillary-like structures were generated upon seeding on Cultrex. Structure analysis was performed using WIMASIS. Total RNA was extracted using TRIzol and RT-qPCR was performed to assess gene expression or screen RT Our findings revealed that in vitro priming of ADMSCs with Cultrex led to the formation of 3D capillary-like structures and the acquisition of an inflammatory molecular signature. VM-derived ADMSCs share a common proinflammatory molecular signature similar to that induced in 2D ADMSC monolayers by tumor necrosis factor (TNF)-alpha and are characterized by upregulated expression of COX2, CCL2, CCL5, CXCL5, CXCL8, IL-6, SNAI1, and MMP9. Interestingly, pharmacological inhibition or gene silencing of the JAK2/STAT3 signaling pathway reduced chemotactic cell migration, in vitro VM and the expression of proinflammatory and invasive biomarkers. Overall, we provide novel evidence that inhibiting JAK2/STAT3-regulated VM can also alter the acquisition of a proinflammatory signature and prevent the contribution of ADMSCs to alternative tumor neovascularization processes. Show less

Voghera pepper (VP) extracts were demonstrated to have anti-oxidant ability in several cell types. This study aimed to assess whether VP-extracts could lower oxidative stress and modulate thyroid cancer (TC) cells behavior Extracts were analyzed using the LC-DAD-MS system. Thyroid cell lines, both normal (NHT) and cancerous (TPC-1 and 8505C) were treated with increasing concentrations of Yellow (YVP) and Green (GVP) VP-extracts over time. Viability and proliferation were assessed in all cell types. Changes in Reactive-oxygen-species (ROS) production by TPC-1 and 8505C were assessed by flow-cytometry. The mRNA expression of anti-oxidant mediators ( Treatment with GVP or YVP reduced the viability of TPC-1 and 8505C but not those of NHT, without effects on cells proliferation. GVP and YVP reduced basal and H This is the first demonstration of the potential beneficial effects of VP extracts in TC in terms of reduction of oxidative stress, increase of antioxidant markers, and modulation of markers of metastasis and de-differentiation in TC cells. Show less

Myocardial fibrosis (MF) is a key pathological process driving heart failure, characterized by excessive extracellular matrix (ECM) deposition and impaired cardiac function. Although myocyte-specific enhancer factor 2 A (MEF2A) is implicated in cardiac fibroblast activation, its role in MF remains unclear. We manipulated MEF2A expression in cardiac fibroblasts (CFs) through knockdown and overexpression, and assessed fibrosis markers, migration, and RhoA signaling. Binding of MEF2A to the Snail1 promoter was predicted using JASPAR and validated by chromatin immunoprecipitation (ChIP) and luciferase reporter assays. Rescue experiments with Snail1 overexpression and RhoA inhibition were performed. An angiotensin II (Ang II)-induced MF mouse model was used to evaluate cardiac function by echocardiography and to assess collagen deposition through picrosirius red (PSR) staining. MEF2A was significantly upregulated in Ang II-induced fibrotic hearts and CFs. MEF2A knockdown reduced α-SMA and Col1a1 expression, inhibited CF migration, and suppressed activation of the Snail1/RhoA/α-SMA pathway. ChIP and luciferase assays confirmed the direct binding of MEF2A to the Snail1 promoter. Inhibition of RhoA signaling reversed MEF2A-induced myofibroblast activation and migration. Rescue experiments showed that Snail1 overexpression restored the fibrotic phenotype suppressed by MEF2A knockdown. In vivo, MEF2A knockdown improved left ventricular function, reduced collagen deposition (PSR staining), and lowered heart weight/tibia length ratios. MEF2A promotes myocardial fibrosis by directly activating Snail1 and engages the RhoA/α-SMA pathway. Targeting MEF2A offers a promising therapeutic strategy to attenuate MF and improve heart function. Show less

In this study, effects of environmental carcinogen benzo[a]pyrene (BaP) on deregulation of sphingolipid (SL) and glycosphingolipid (GSL) metabolism were studied during BaP-induced transformation of normal human bronchial epithelial HBEC-12KT cells. After 2-weeks of exposure, BaP altered their morphology, while it downregulated sphingosine-1-phosphate (S1P) and upregulated sphingosine, gangliosides, GM3 and Lc3 GSLs. A longer, 8-week exposure to BaP, further increased cell migratory capacity, induced epithelial-to-mesenchymal transition (EMT) markers and EMT-related transcriptional regulators (SNAI1, ZEB1 and ZEB2), and it increased intracellular sphingosine, ceramide-1-phosphate, as well as a series of GSLs (glucosylceramide, lactosylceramide, GM1a, GD3, Lc3 and Gb3). A distinct profile of SL/GSL levels was observed in fully transformed cells established via exposure to BaP for 12 weeks. Increased sphingosine, S1P, ceramide-1-phosphate, GD3, Lc3 and Gb3 levels were paralleled by a decrease of other SL/GSLs, including GM3 pathway. These alterations were also partly reflected within extracellular vesicles and microvesicles, particularly in those released from fully transformed cells. Significantly altered enzymes of SL/GSL metabolism included a downregulation of S1P lyase and increased S1P, downregulation of GM-synthetic enzymes, and upregulation of enzymes of GD3 and Gb3 synthesis. Using siRNA-mediated knockdown of individual EMT transcriptional regulators, we then found them to play only a partial role in regulation of S1P, GM, GD3, Lc3 or Gb3, via deregulation of expression of the respective enzymes, suggesting that their enzymatic activities can be regulated also by other mechanisms. The mechanisms underlying deregulation of SL/GSL levels elicited by carcinogenic environmental pollutants and functions of individual SL/GSL species deserve further attention. Show less

Renal cell carcinoma (RCC) has a well-established propensity to form grossly visible tumour thrombi; however a comprehensive understanding of the underlying mechanisms is still lacking. The epithelial-mesenchymal transition (EMT) has been implicated in the progression of many carcinomas, including RCC; however, its exact role in the formation of venous tumour thrombi remains unclear. This study aims to explore the involvement of the EMT in venous invasion in RCC. In 14 patients with WHO/ISUP grade 2/3 clear cell RCC with venous invasion, the expression of main EMT markers (the Show less

Endothelial-to-mesenchymal transition (EndMT) is a biological process through which lung vascular endothelial cells (ECs) transdifferentiate into mesenchymal-like cells. EndMT has recently been implicated in the development and progression of pulmonary vascular remodeling in pulmonary hypertension (PH); however, its underlying regulatory mechanisms remain incompletely understood. MicroRNAs (miRNAs) are key post-transcriptional regulators of EC gene expression and cellular responses to various stimuli. Notably, microRNA-153 (miR-153) has been shown to directly target SNAI1 to modulate epithelial-to-mesenchymal transition (EMT), a process closely related to EndMT and extensively studied in cancer. Whether miR-153 also participates in EndMT regulation, however, remains unknown. In this study, we demonstrate that 72-hour hypoxic exposure induces SNAI1-mediated EndMT in human lung vascular ECs. Hypoxia also increased cell proliferation and disrupted intercellular junctions, leading to enhanced endothelial permeability. Reduced miR-153 expression was observed in both hypoxia- and TGF-β1-induced EndMT, as well as in ECs isolated from PH patients exhibiting an EndMT phenotype. Similar to hypoxia, TGF-β1 promoted EC permeability. Loss of miR-153 enhanced SNAI1-mediated EndMT, endothelial survival, and permeability under normoxic conditions, whereas miR-153 overexpression attenuated EndMT induced by hypoxia or TGF-β1. However, miR-153 restoration did not completely recover endothelial barrier integrity disrupted by these stimuli. In conclusion, miR-153 serves as a critical regulator of EndMT, maintaining endothelial identity and barrier function. Therapeutic delivery of miR-153 may therefore represent a novel strategy to inhibit EndMT and attenuate pulmonary vascular remodeling in PH. Show less

The development of an immunosuppressive microenvironment is a critical factor in stomach carcinogenesis. Polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) serve a pivotal function in mediating immune suppression. However, the precise mechanisms underlying PMN-MDSCs infiltration into the tumor immune microenvironment (TIME) and their immunosuppressive functions remain poorly understood. In this investigation, we observed that PMN-MDSCs were up-regulated during stomach carcinogenesis, with gastric cancer (GC) cells secreting CCL26 to promote the infiltration of PMN-MDSCs into the TIME via the CX3CR1 receptor. The infiltrating CX3CR1 Show less

The widely accepted theory of endometriosis posits that endometriosis stems from the translocation of endometrial tissue through the fallopian tubes into the abdominal cavity. However, the exact pathogenesis and critical molecules of endometriosis remain unclear. Here, we find that alanyl-tRNA synthetase 1 (AARS1) is abundantly expressed in endometrial tissues and promotes the proliferation, migratory capability, and invasive potential in endometriotic stromal cells (EESC) and 11Z cells. Moreover, AARS1 enhances epithelial-to-mesenchymal transition in EESC and 11Z cells. In addition, AARS1 could lactylate Snail1 to maintain its protein stability. In summary, this work identifies a crucial role of AARS1 in advancing endometriosis, which may provide new insights into its pathogenesis and future disease management. Show less

Despite extensive research, the pathogenesis and predispositions underlying long COVID (long-term coronavirus disease 2019) remain poorly understood. To address this, we analyzed the immunological landscapes of 44 patients with long COVID and 44 matched convalescents using single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) and validated the findings with plasma cytokine measurements via Luminex technology. While the immune cell compositions showed minimal quantitative differences only among natural killer (NK) cells, the transcriptome analyses identified distinct gene expression patterns, particularly in classical monocytes: patients with long COVID exhibited downregulation of the inflammation-associated genes, including These findings show that monocytes might be dysregulated and/or exhausted in patients with long COVID. Show less

Snail and Zeb1 have been suggested as markers for the hybrid/mixed epithelial (E)/mesenchymal (M) state of cancer cells. Such cancer cells co-express E- and M-specific transcripts and possess cancer stem cell properties. M13HS-2/-8 tumor hybrid clones derived from human M13SV1-EGFP-Neo breast epithelial cells and human HS578T-Hyg breast cancer cells exhibited co-expression of Snail and Zeb1. To explore the impact of Snail on stemness/epithelial-to-mesenchymal transition (EMT)-related properties in M13HS-2/-8 tumor hybrid clones, Snail was knocked out (KO) using CRISPR/Cas9. Mammosphere formation, colony formation, Western blot analyses, cell migration, and invasion assays were conducted for the characterization of Snail knockout cells. Interestingly, Snail-KO in M13SV1-EGFP-Neo cells resulted in the up-regulation of vimentin and N-cadherin, suggesting EMT induction, which was associated with a significantly enhanced colony formation capacity. In contrast, EMT marker pattern and colony formation capacities of M13HS-2/-8 Snail-KO tumor hybrid clones remained unchanged. Notably, the mammosphere formation capacities of M13HS-2/-8 Snail-KO tumor hybrid clones were significantly reduced. The migratory behavior of all Snail-KO cells was not altered compared with their wild-type counterparts. In contrast, M13HS-2 hybrids and their M13HS-2 Snail-KO variant exhibited a markedly enhanced invasive capacity. Therefore, Snail plays a role as a mediator of stemness properties rather than mediating EMT. Show less

The majority of pregnancy loss in ruminants occurs during the first two months of gestation, and a failure in placenta development is a major cause of pregnancy loss in cattle after day 20. Gaining a cell-type level understanding of normal placental development is essential for uncovering how this critical organ, responsible for nutrient exchange, gas transfer, and waste removal, fails during pregnancy loss. This study integrated single-cell RNA sequencing (scRNA-seq) from sheep and cattle during early placental development. Nineteen distinct cell populations were identified across species, with mesenchymal, epithelial, and trophoblast cells showing largely conserved expression profiles. Interestingly, two trophoblast clusters were unique to cattle, with one expressing IFNT2 (uninucleate) and another expressing CSH2 and PAG17 (binucleate). Genes associated with epithelial-to-mesenchymal transition (EMT), such as SNAI1, SNAI2, ZEB1, VIM, CDH1, and CLDN4, showed dynamic and prominent expression patterns in trophoblasts. Pseudotime and cell-cell signaling analyses supported the occurrence of EMT in uninucleate trophoblasts. Gene ontology comparisons revealed similarities between ruminant and human extravillous trophoblasts, suggesting conserved EMT across placental types. Collectively, these findings highlight EMT as a potentially critical process in early ruminant placentation. Show less

Oral squamous cell carcinoma (OSCC) remains a challenging malignancy with poor 5-year survival rates due to diagnosis at an advanced stage and a high likelihood of recurrence and metastasis. These aggressive traits may be influenced by cancer stem cells (CSC) and epithelial-mesenchymal transition (EMT). This study investigated the prognostic significance of the CSC marker CD44 and EMT-related proteins (Snail1, Snail2, E-cadherin, N-cadherin) in 132 OSCCs using immunohistochemistry. The comprehensive survival analysis included univariate and multivariate (stepwise method) Cox regression for disease-specific survival (DSS) and disease-free survival (DFS), Kaplan-Meier curves based on log-rank testing, and receiver operating characteristic (ROC) analysis to assess the predictive accuracy of the markers. High CD44 expression independently predicted worse DSS (HR = 2.74, 95% CI 1.44-5.23, p = 0.003) and DFS (HR = 2.22, 95% CI 1.16-4.23, p = 0.01), and Snail1 was significantly associated with poor DSS (HR = 2.62, 95% CI 1.37-5.03, p = 0.004). The combined expression of CD44 and Snail1 improved the discrimination of worse outcomes compared to markers individually. The presence of lymphovascular invasion (HR = 8.68, 95% CI 3.81-19.75, p < 0.0001) and a positive surgical margin (< 5 mm; HR = 4.45, 95% CI 1.99-9.96, p = 0.0003) were also independently associated with DSS. The results of this study highlight the prognostic significance of CD44 and Snail1 in OSCC, emphasizing their potential interplay in tumor aggressiveness. Show less

Gastric cancer is aggressive with poor prognosis due to high invasion and metastasis rates, a hallmark of cancer. The Snail family (SNAI1 and SNAI2) drives EMT, enabling epithelial cells to gain migratory and invasive traits. We used "limma" package to identify genes with differential expression between high and low levels of SNAI1/SNAI2 in TCGA stomach adenocarcinoma dataset, intersecting these with cancer invasion and metastasis genes obtained from 5 databases. Using Cox regression analysis, we developed a risk score model and created a nomogram incorporating clinical data. The model's prognostic accuracy was validated with survival and ROC analyses in both TCGA and GEO datasets. Additionally, we performed WGCNA and constructed a ceRNA network to investigate gene interactions, and used CIBERSORT analysis to evaluate immune cell composition in the tumor microenvironment. We developed 5 and 9 risk signatures and nomograms incorporating clinical data. Survival analysis showed high-risk patients had worse overall survival than low-risk patients. WGCNA identified a lightyellow module associated with SNAI1 and SNAI2 expressions, emphasizing extracellular matrix organization. CeRNA network analyses found 6 common hub genes linked to SNAI1 and SNAI2. Immune profiling showed that SNAI1 expression was related to 8 types of immune cells, while SNAI2 was connected to 6, indicating their roles in influencing the tumor microenvironment. This study highlights the significant prognostic impact of SNAI1 and SNAI2 in stomach adenocarcinoma, linking their high expression to poorer survival and aggressive tumor behavior, while also identifying potential therapeutic targets through comprehensive computational analysis. Show less

Melanoma cells evade drug treatment by changing their phenotype from proliferative to migrative cells and vice versa in a process known as phenotype switching. The Microphthalmia-associated transcription factor (MITF) is a key regulator of phenotype switching in melanoma. Previous studies have shown that loss of MITF affects the expression of epithelial-to-mesenchymal transition marker genes such as E-cadherin (CDH1) and N-cadherin (CDH2). However, the specific roles of CDH1 and CDH2 in phenotype switching as well as their direct correlation with MITF remain unclear. This study aimed to investigate how MITF regulates CDH1 expression in melanoma. The results showed that a 1 kb intronic CDH1 fragment (CDH1-B) leads to MITF-dependent activation of CDH1 expression through specific binding sites. Although MITF represses the expression of the epithelial-to-mesenchymal transition transcription factors SNAIL, ZEB1, and TWIST1, knockdown of SNAI1 and TWIST1 did not affect CDH1 expression or expression from the CDH1-B element. In addition, ZEB1 did not affect expression from the CDH1-B element, suggesting that MITF activates CDH1 directly through this regulatory element. Our results show the direct role of MITF in regulating CDH1 expression in melanoma, highlighting an important step in the phenotype switching process. Show less

Rheumatoid arthritis (RA) is one of the most common chronic inflammatory autoimmune diseases, and ferroptosis has been associated with its pathogenesis. TRIM16 belongs to the TRIM protein family and possesses various biological function. However, the role of TRIM16 in RA has not been reported. Our results showed that TRIM16 was upregulated in collagen-induced arthritis (CIA) mice, and TRIM16 overexpression alleviated joint inflammation. Notably, the level of 4-HNE was decreased in CIA mice, whereas TRIM16 overexpression restored it. The expression of GPX4 and SLC7A11 was upregulated in CIA mice, whereas TRIM16 overexpression significantly suppressed their levels, suggesting that TRIM16 promotes ferroptosis. We then detected TRIM16 expression in TNF-α-induced fibroblast-like synoviocytes (FLS), and found that TNF-α stimulation reduced TRIM16 expression. Overexpression of TRIM16 increased the lipid ROS, Fe Show less

Biliary atresia (BA) is a severe pediatric liver disease in which progressive liver fibrosis (LF) significantly affects the prognosis. Epithelial-mesenchymal transition (EMT) is considered a key factor in the development and progression of LF. This study aimed to investigate the role and mechanism of PEAK1-related kinase activating pseudokinase 1 (PRAG1) in the EMT-related LF process in BA. We found that the expression of PRAG1 was significantly elevated in both patients with BA and the bile duct ligation (BDL) model, and predominantly localized on biliary epithelial cells. Also, the expression of PRAG1 positively correlated with the cholangiocyte marker KRT19 and the mesenchymal marker ACTA2, and increased with the severity of fibrosis. In human intrahepatic biliary epithelial cells (HIBECs), PRAG1 promoted the expression of mesenchymal markers (VIM and ACTA2) and fibrosis markers (COL1A1 and FN1), inhibited the expression of the epithelial marker CDH1, and enhanced cell proliferation. The key factor of EMT-SNAIL1 presented increased expression and delayed degradation after overexpression of PRAG1. Moreover, we identified PRAG1 could bind with F-box protein 11 (FBXO11) and subsequently reversed FBXO11-mediated inhibition of SNAIL1 protein expression, cell proliferation, and the EMT phenotype. This study provides the potential role of PRAG1 in the mechanisms underlying the LF progression of BA. Show less

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide, with tumor progression often driven by dysregulated oncogenic pathways. USP6NL, a known regulator of endocytic trafficking, has recently been implicated in tumorigenesis. However, its precise role in CRC remains unclear, and more studies are still needed to deepen our understanding of underlying mechanisms implicated in its oncogenic role. Therefore, silencing USP6NL could provide a novel therapeutic strategy by concurrently disrupting several oncogenic mechanisms, creating a new avenue for CRC management, particularly in patients who develop resistance against conventional therapies. This study investigates the impact of USP6NL knockdown on CRC cell morphology, proliferation, apoptosis, migration, angiogenesis, and metabolic adaptation, providing mechanistic insights into its oncogenic functions. HCT116 colorectal cancer cells were transfected with USP6NL-specific siRNA. Immunocytochemistry was used to confirm successful silencing, functional assays were performed to assess changes in cell morphology using phase-contrast and scanning electron microscopy, and colony formation and wound healing assays were performed to assess cell clonogenic capacity and migration, respectively, in addition to apoptosis assay via flow cytometry, and RT²-Profiler PCR array to measure variation in gene expression of 84 cancer-related genes. Statistical analyses were performed to evaluate significant differences between control and USP6NL-silenced groups. USP6NL depletion led to profound morphological changes, including membrane blebbing, cell shrinkage, and loss of adhesion, reflecting late apoptotic features of cells. These findings were further supported by flow cytometry, which confirmed increased apoptosis, with a higher proportion of late apoptotic cells (20.99% in USP6NL knockdown vs. 2.69% in control, p = 0.042). Colony formation assays revealed a significant reduction in the clonogenic potential, suggesting a critical role of USP6NL in promoting CRC cell proliferation (p ≤ 0.05). The wound healing assay demonstrated impaired migration in USP6NL-silenced cells, with a marked delay in wound closure (p = 0.0201 at 48 h). Gene expression analysis revealed a significant downregulation of VEGFC (-8.62-fold) and ANGPT2 (-4.03-fold), impairing angiogenesis and suppressing FOXC2, SNAI1, and SNAI2, indicating EMT inhibition. Additionally, CASP9, APAF1, and BCL2L11 were upregulated, confirming the activation of intrinsic apoptosis, while metabolic regulators HIF1A and LDHA were downregulated, suggesting impaired tumor hypoxic adaptation. This study establishes USP6NL as a key modulator of CRC progression, regulating proliferation, apoptosis, migration, angiogenesis, and metabolic pathways. The loss of USP6NL leads to EMT suppression, apoptosis induction, and reduced tumor cell viability, positioning it as a potential therapeutic target in colorectal cancer. Further investigations are warranted to explore USP6NL's interactions in oncogenic signaling networks and its feasibility as a target for CRC therapy. It could serve as a promising therapeutic target in colorectal cancer, potentially enhancing tumor cell death and limiting metastasis. Targeting USP6NL could also provide a novel approach in combination with existing therapies, improving treatment efficacy and reducing side effects. Show less

Preventing the progression from acute kidney injury (AKI) to chronic kidney disease (CKD) remains a considerable clinical challenge. In this study, we elucidate the role of WNT5A in accelerating the AKI-to-CKD transition and its underlying mechanisms. Renal biopsies from patients with AKI showed marked upregulation of WNT5A and its receptor, CD146, in proximal tubules, with higher expression in patients with CKD progression. In murine AKI models, Wnt5a knockdown attenuated CKD progression. Conversely, proximal tubular overexpression of Wnt5a exacerbated renal fibrosis in ischemia-reperfusion injury (IRI) mice, which was alleviated by Box5, a specific WNT5A antagonist. In vitro, WNT5A overexpression in transforming growth factor β (TGF-β)-stimulated HK-2 cells promoted CD146 upregulation, activated JNK phosphorylation, and enhanced SNAI1 expression. The genetic silencing of WNT5A/CD146 and JNK inhibition suppresses SNAI1 expression and attenuates fibrotic responses. Mechanistically, JNK-mediated c-JUN phosphorylation promoted its interaction with KLF5 at the SNAI1 promoter, driving renal fibrosis. Elevated serum levels of soluble CD146 correlated with renal function in patients with AKI and were higher in patients exhibiting CKD progression. Inhibition of WNT5A could serve as a therapeutic target for delaying renal fibrosis in AKI progression. Show less

Transforming growth factor-β (TGF-β) plays well-established roles in cancer cell invasion and epithelial-mesenchymal transition (EMT); however, its role in thyroid carcinoma (TC) remains unclear. This study aimed to evaluate the effects of TGF-β on EMT in TC and determine its underlying mechanisms. Treatment of TC cell lines with TGF-β the morphology of thyroid cancer cells changed, Immunofluorescence staining revealed that the localization of E-cadherin shifted from the cell membrane to the cytoplasm, and the fluorescence intensity decreases. Wound-healing assay in BCPAP and TPC-1 revealed that migration ability was significantly higher in the TGF-β (5 ng/mL) treatment group than in the control group (P < 0.01). Transwell assays showed that the invasive abilities of TGF-β-treated BCPAP, TPC-1, and K1 cells were 7-, 10-, and 6-fold higher than those of the control group, respectively (P < 0.05). After TGF-β treatment, mRNA levels of SNAI1 significantly increased in TPC-1 and BCPAP cell lines. Treatment of TC cell lines with TGF-β downregulated the epithelial marker E-cadherin and upregulated the mesenchymal markers N-cadherin and vimentin, at the mRNA level. Western blotting indicated similar results at the protein level, TSH could enhance this process. TGF-β promotes EMT-like phenotypic changes in thyroid cancer cells, accompanied by upregulation of SNAI1 and EMT-related markers, which is enhanced by TSH. Overall, this study provides a basis for the development of therapeutic strategies for TC targeting the EMT. Show less

Breast cancer is considered to be the most common cancer that affects women worldwide, where it accounts for approximately 38.8% of all cancer cases among females. Luminal subtypes are the most prevalent in Egypt. Small noncoding RNAs also called microRNAs (miRNAs) influence gene expression posttranscriptionally. Since they regulate the epithelial-mesenchymal transition process, which is vital for tumor invasion and metastasis, microRNAs play a critical role in the progression of cancer. This study has investigated the expression profiles of four microRNAs (miR-101-3p, miR-106a-5p, miR-106b-5p, and miR-130b-5p) and their impacts on genes associated with epithelial-mesenchymal transition (EMT) in luminal breast cancer. Tissue samples from 43 luminal breast cancer patients and 18 controls have been studied via real-time PCR (RT-qPCR). The association between the expression levels was evaluated using the Pearson correlation test. The correlation between the measured variables and numerous clinicopathological characteristics was assessed using the linear regression test. The results demonstrated that miR-101-3p, miR-106a-5p, and miR-106b-5p were significantly dysregulated, highlighting their possible role as oncogenes or tumor suppressors in the development of breast cancer. EMT markers, especially Twist, SNAI1, and E-cadherin, show significant alterations, indicating the activation of EMT pathways in luminal breast cancer. Correlation analysis showed interactions between miRNAs and EMT-related genes, showing a negative correlation between miR-101-3p and SNAI1, as well as a positive correlation between Twist and miR-106a-5p. Moreover, logistic regression analysis associated expression levels of those miRNAs with clinicopathological characteristics, such as body weight, age, and tumor laterality. These findings highlight the leading role of miR-101-3p and miR-106b-5p in the progression of luminal breast cancer via interacting with the EMT process and their potential as diagnostic, prognostic, and therapeutic targets. Show less

The precise involvement of Guanine Nucleotide-Binding Protein-Like 3-Like Protein (GNL3L) in lung cancer progression and invasion remains unclear. In this study, we explored the impact and underlying mechanisms of GNL3L on the proliferation, migration, and invasion of lung adenocarcinoma (LUAD), and evaluated the therapeutic potential of targeting GNL3L. Inhibition of GNL3L expression led to a notable decrease in the in vitro proliferation, migration, and invasion of A549 and H1299 non-small cell lung cancer (NSCLC) cells. Meanwhile, GNL3L silencing could significantly reduce the tumor volume of the nude mice and improve the outcomes of tumor-bearing mice in vivo. Additionally, inhibition of GNL3L expression dramatically suppressed NF-κB activation and Slug, MMP2, and MMP9 expression. Overexpression of Slug or treatment of the GNL3L-deficient cells with NF-κB activator can partially restore the growth suppressed by GNL3L deficiency, and combined treatment with Slug overexpression and NF-κB activator could totally restore the suppressed cell growth caused by GNL3L deficiency. Moreover, the overexpression of MMP2 or MMP9 could partially enhance the reduced migration and invasion caused by GNL3L deficiency, and this GNL3L-deficiency-caused suppression of migration and invasion can be totally restored by the overexpression of MMP2 and MMP9 together. These results strongly indicated that GNL3L has the capability to activate the NF-κB and increase Slug, MMP2, and MMP9 expression, which in turn could stimulate the proliferation, migration, and invasion of lung cancer cells. NF-κB activation and Slug, MMP2, and MMP9 expression enhanced by GNL3L, leading to the promotion of proliferation, migration, and invasion of lung cancer cells, indicating the therapeutic implications and potential significance of these pathways in the progression and invasion of NSCLCs that overexpress GNL3L protein. Show less

The IARC classified betel nut as Group 1 carcinogen (2004) and arecoline as Group 2B carcinogen (2020), with approximately one-third of global oral cancer cases attributed to smokeless tobacco or betel nut consumption. While current evidence establishes an association between arecoline and oral cancer, the underlying molecular mechanisms remain complex and poorly elucidated. This study employs network toxicology integrated with molecular docking techniques to systematically investigate the potential molecular pathogenesis of arecoline-induced oral cancer, aiming to provide novel insights for targeted therapeutic strategies. The SMILES structure of arecoline was retrieved from PubChem for foundational data preparation. Toxicity profiling was conducted using ProTox-3.0 and ADMETlab databases. Potential targets of arecoline were identified via STITCH and SwissTargetPrediction. Oral cancer-related targets were collated from GeneCards, OMIM, and TTD. Intersection analysis between arecoline targets and oral cancer-associated targets was performed to identify shared targets, which were further utilized to construct compound-target regulatory network and subjected to PPI, GO, and KEGG analyses. Core targets driving oral cancer were screened using the cytoHubba plugin. Then, the correlation between core targets and immune cell infiltration in oral cancer was explored, and molecular docking validated the binding affinity of arecoline to core targets. Finally, Gromacs 2022.3 software was used to simulate the molecular dynamics of the complexes obtained by molecular docking for 100 ns. Using the STITCH and SwissTargetPrediction databases, a total of 46 potential targets of arecoline were identified. Concurrently, 2,375 oral cancer-related targets were retrieved from GeneCards, OMIM, and TTD. Intersection analysis of these two target sets yielded 26 overlapping targets. PPI analysis revealed that TP53, IL6, SNAI1, and CASP3 occupied central positions in the network, exhibiting extensive interactions with other target proteins. Enrichment analysis comprehensively elucidated the molecular functions, biological processes, cellular components, and associated pathways of these overlapping targets. Further screening using Cytoscape software identified four core targets: TP53, TNF, IL6, and CASP3. Immune infiltration analysis indicated that the expression levels of TP53, TNF, IL6, and CASP3 in oral cancer tissues were positively correlated with the infiltration levels of immune cells, including CD8 + T cells, Th1 cells, NK cells, and macrophages. Molecular docking experiments demonstrated strong binding activities between arecoline and TP53, IL6, and CASP3, while TNF also exhibited moderate binding affinity. Dynamic simulation further verified the stable binding of arecoline to TP53, TNF, IL6 and CASP3. Arecoline may induce oral cancer by acting on core targets including TP53, TNF, IL6, and CASP3, which interfere with normal cellular growth regulation, inflammatory responses, and apoptotic mechanisms. Therapeutic strategies targeting TP53, TNF, IL6, and CASP3 may represent novel research directions for clinical diagnosis and treatment of oral cancer. Show less