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Numerous studies have shown that exposure to cadmium [Cd(II)] contributes to the development of cancers in the lung and other organs. Cd(II) compounds are classified as confirmed human carcinogens; however, the mechanisms underlying Cd(II)-induced carcinogenesis remain poorly understood. Small nucleolar RNA host gene 1 (SNHG1), a long non-coding RNA (lncRNA), has been identified as an oncogene. In this study, we investigated the role of SNHG1 in the invasion and migration of Cd(II)-transformed cells. Our findings revealed that SNHG1 expression was significantly elevated in Cd(II)-transformed cells compared to their passage-matched normal BEAS-2B counterparts. Silencing SNHG1 reduced the invasive and migratory capacities of Cd(II)-transformed cells and inhibited malignant transformation induced by long-term Cd exposure. Notably, ectopic expression of SNHG1 alone in BEAS-2B cells was sufficient to drive malignant transformation and enhance invasion and migration, underscoring its oncogenic potential. SRY-box 2 (Sox2), a transcription factor implicated in cancer cell proliferation, invasion, and migration, was found to be upregulated in Cd(II)-transformed cells, while SNHG1 knockdown led to decreased Sox2 protein levels. Similarly, ras-related C3 botulinum toxin substrate 1 (Rac1), a key regulator of cytoskeletal dynamics linked to tumor growth, invasion, and metastasis, was also elevated in Cd(II)-transformed cells. Knockdown of SNHG1 reduced Rac1 protein levels, and Rac1 knockout significantly suppressed invasion and migration. Additionally, we observed increased expression of Slug, a key transcription factor invovlved in epithelial-mesenchymal transition (EMT), and decreased expression of its downstream target E-cadherin in Cd(II)-transformed cells. Collectively, these results demonstrate that elevated SNHG1 promotes the expression of Sox2, Rac1, and Slug, thereby driving the invasive and migratory behavior of Cd(II)-transformed cells. Show less

Abnormal zygotic genome activation (ZGA) during the early development of somatic cell nuclear transfer (SCNT) embryos is one of the main reasons for the low cloning efficiency. The double homeobox (DUX) family, which includes important transcription factors in mammals, has been shown to play an important role in the ZGA process in mice. However, the role of DUXA, a member of the DUX family, in the early development of porcine somatic cloned embryos is unknown. Here, CRISPR/Cas9 gene editing and lentiviral infection technologies were used to construct stable DUXA knockout and overexpression cell lines for the production of SCNT embryos. Compared with that of wild-type (WT) SCNT embryos, the blastocyst rate of DUXA knockout embryos was significantly lower (P < 0.05), whereas the blastocyst rate of DUXA-overexpressing embryos was significantly greater (P < 0.05). Moreover, RT‒qPCR results revealed that DUXA knockout significantly reduced the expression levels of ZGA-related genes (TDG, SNAI1, RSRP1, TFAP2C, ZSCAN4, LEUTX, and KLF17) (P < 0.05). Additionally, in DUXA-overexpressing embryos, the mRNA levels of TDG, SNAI1, RSRP1, and TFAP2C significantly decreased (P < 0.05), whereas the ZSCAN4, LEUTX, and KLF17 mRNA levels increased (P < 0.05). These findings suggest that DUXA regulates the early development of porcine SCNT embryos by modulating the expression of ZGA-related genes. This research provides significant insights into the potential mechanisms of early embryo loss in porcine SCNT. 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

Although it has been shown that the tumor extracellular matrix (ECM) may sustain the cancer stem cell (CSC) niche, its role in the modulation of CSC properties remains poorly characterized. To elucidate this, paired tumor and adjacent normal mucosa, derived from colon cancer patients' surgical resections, were decellularized and recellularized with two distinct colon cancer cells, HT-29 or HCT-15. Methods: The matrix impact on cancer stem cell marker expression was evaluated by flow cytometry and qRT-PCR, while transforming growth factor-β (TGF-β) secretion and matrix metalloprotease (MMP) activity were quantified by ELISA and zymography. Results: In contrast to their paired normal counterparts, the tumor decellularized matrices enhanced HT-29 expression of the pluripotency and stemness genes 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

Endothelial-to-mesenchymal transition (EndMT) induced by dysfunctional pulmonary artery endothelial cells (PAECs) is regarded as an initiating and pivotal factor in pulmonary hypertension (PH). This study focuses on identifying a novel therapeutic target for regulating EndMT in PH. A comprehensive analysis of 2 hypoxic PAECs datasets yielded 310 overlapping upregulated and 229 downregulated differentially expressed genes (DEGs). These upregulated DEGs were primarily enriched in HIF-1 signalling pathway and glycolysis/gluconeogenesis, while downregulated only in spliceosome, as indicated by KEGG. Through PPI network analysis and the application of MCC algorithms, 5 hub genes were identified among these upregulated DEGs: GAPDH, LDHA, ALDOA, PFKL, and PFKP. Their enrichment in the 2 aforementioned pathways was confirmed by cross-pathway DEGs analysis and ClueGo. Among the hub genes, LDHA was chosen as the key gene based upon expression and correlation analysis of the validation set from PH patients. Subsequent GSEA also revealed the enrichment of LDHA in these 2 pathways. Additionally, the increased expression of LDHA protein in tissues and cells was confirmed, and the elevated enzymatic activity of LDHA in clinical serum samples was also verified. From 2 online databases, 4 LDHA inhibitors were filtered out, and the stable binding between the inhibitors and the LDHA protein was confirmed through molecular docking and molecular dynamics simulation. Finally, the experimental results indicated that one of the inhibitors FX11 reversed EndMT by inhibiting the lactate-SNAI1 axis, thereby alleviating hypoxia-induced PH. The potential of LDHA as a therapeutic target for PH by modulating EndMT was proposed in this study. Show less

Anoikis is a new mode of cell death that has been shown to correlate significantly with tumors. However, the clinical prognostic significance of anoikis in lung squamous cell carcinoma (LUSC) remains poorly studied. The differentially expressed ARGs and candidate genes were selected by the differential analysis to construct a predictive model. Independent prognostic gene was determined by Cox and LASSO analysis and we used the HCC95 and NCI H520 cell line to verify the gene function. We used the data from TCGA, GEO, GeneCards, and Harmonizome databases to analyze the immune microenvironment, functional enrichment, and drug sensitivity analysis. We identified 717 differentially expressed and selected 3 ARGs (FADD, SNAI1, and BAG4) to construct a predictive model. We found that SNAI1 is an independent prognostic gene and confirmed that knocking out the SNAI1 inhibited the HCC95 We used ARGs to construct a prognosis model for LUSC that can accurately predict the prognosis of LUSC patients. ARGs, especially SNAI1, play an essential role in developing LUSC. These findings could provide individualized treatment plans and new research ideas for LUSC patients. Show less

HNSCC is a highly aggressive cancer of the head and neck region, and there is an urgent need to find novel potential targets for its diagnosis and treatment. Long non-coding RNAs (lncRNAs) have emerged as important therapeutic and diagnostic targets for multiple cancers, including HNSCC. LINC01518 promotes the proliferation of oesophageal cancer cells, but the involvement of LINC01518 in HNSCC pathophysiology is unknown. We show that LINC01518 expression is significantly upregulated in high-grade HNSCC tumor samples in comparison to normal tissue, and transforming growth factor- β (TGF-β) promotes LINC01518 expression in HNSCC cell lines. Loss-of-function studies suggest that LINC01518 promotes cell proliferation, migration, and invasion in HNSCC cells. In addition, LINC01518 depletion sensitizes HNSCC cells to cisplatin-mediated apoptosis. Mechanistically, LINC01518 acts as a competitive endogenous RNA and binds to miR-1-3p and miR-216b-5p, resulting in up-regulation of their target genes Slug and GRP78, respectively. Our findings suggest that LINC01518 is an attractive therapeutic target for HNSCC. 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

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

HUWE1, a member of HECT E3 ubiquitin ligase family, is implicated in a variety of cellular processes. Recent studies find that HUWE1 also plays critical roles in germ cell development and inactivation of HUWE1 causes germ cell loss in both male and female mice. In this study, we found that Huwe1 was also highly expressed in testicular Sertoli cells. Inactivation of Huwe1 in Sertoli cells resulted in loss of cell polarity, which in turn caused germ cells loss and male infertility. Further study revealed that dysregulation in the expression of cytoskeletal and adhesion-related molecules, as well as a significant increase in EMT-related trans-factors SNAI1&2 in Huwe1-deficient Sertoli cells. Intriguingly, the protein level of WT1 was significantly increased in Huwe1-deficient Sertoli cells, and overexpression of Wt1 in Sertoli cells also caused the defects in spermatogenesis which was consistent with Huwe1 CKO mouse model. Furthermore, the defect of spermatogenesis in Huwe1 CKO mice was partially rescued by deleting one allele of Wt1 gene. Mechanistic studies revealed that WT1 interacts with HUWE1 protein and it could be ubiquitinated by HUWE1. Our study demonstrates that HUWE1 is involved in the establishment of Sertoli cell polarity mainly by regulating the protein level of WT1 gene. Show less

For workers in the industry, occupational exposure to indium compounds induces pulmonary disorders, such as interstitial pneumonia. Moreover, lung cancer has been reported in both humans and rodents exposed to indium compounds by inhalation. However, the biological mechanism underlying indium-induced disorders is poorly understood. Epithelial-mesenchymal transition (EMT)-the cellular process of losing epithelial and acquiring mesenchymal characteristics-is linked to fibrosis and cancer progression. Therefore, we examined whether indium exposure elicits EMT in vitro. A549 human alveolar epithelial cells treated with indium chloride at doses of 0-500 μg/mL for 24 h were used to analyze EMT marker expression and cytoarchitecture. Significant downregulation of CDH1 mRNA expression as an epithelial marker after treatments at 125, 250, and 500 μg/mL occurred dose-dependently; conversely, the mesenchymal marker SNAI1 was upregulated. Consistent with mRNAs, the expression levels of EMT marker proteins (i.e., E-cadherin, ZO1, SNAIL, and Vimentin) were changed significantly by treatment. While NF-κB signaling was activated in treated cells, indium-dependent changes of CDH1 and SNAI1 mRNA expression were not affected by BAY 11-7082, an NF-κB inhibitor, suggesting that NF-κB activation may be dispensable for indium-induced EMT. Fibroblast-like morphological characteristics, such as actin stress fiber formation and cell elongation, along with deconstruction of cell-cell adhesion complexes, were observed in treated cells. Overall, our study is the first to demonstrate that EMT is caused by indium compounds. This will contribute biologically to understanding the mechanism of EMT induction and clinically to unveiling the pathophysiology of indium lung disease. Show less

Apart from biochemical signals, tumour cells respond to biophysical and mechanical cues from their environment. The mechanical forces from the tumour microenvironment could be in the form of shear stress, tension, or solid stress compression. In this study, we explore the effects of solid stress compression on tumour cells. Solid stress compression, a prevalent biomechanical stimulus accumulated during tumour growth, has been shown to enhance invasive and metastatic phenotypes in cancer cells. However, the underlying molecular mechanism that elicits this aggressive metastatic phenotype, especially in breast cancer, is not extensively studied. Using an established 2D Show less

Apigenin, found in a variety of vegetables and fruits, exhibits anti-oxidant, anti-inflammatory and anticancer effects. Recently, we reported the possibility that apigenin induces apoptosis in human lung adenocarcinoma A549 cells through the miR-34a-5p/SNAI1/caspase-3/-7 pathway. Understanding how apigenin triggers apoptosis in cancer cells will help lay the groundwork for developing effective cancer treatments. The lung adenocarcinoma A549 Cell line was used. To determine whether caspase-8 or caspase-9 is activated, we performed a caspase activity assay. Real-time qRT-PCR was performed to identify mRNAs that may stimulate the upstream pathways, including tumor necrosis factor-a (TNF-a), spondin-2 (SPON2), and interferon-a2 (IFNA2), which are known to be involved in apoptosis in various cancer cell lines. In apigenin-treated cells, early-stage apoptosis was observed at 24 h, with increased activity of caspase-8 at 18 h and again at 24 h, and caspase-9 at 24 h and further at 48 h. However, mRNA levels of caspase-8 and caspase-9 significantly decreased after 24 h. Real-time RT-qPCR analysis revealed increased mRNA levels of TNF-a, spondin-2, and interferon-a2 after 24 h of apigenin treatment in A549 cells, whereas treatment for 48 h led to decreased expression of SPON2 and IFNA2. Apigenin promotes apoptosis in A549 cells by modulating various signaling pathways at different time points. 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

This study aimed to investigate the role of the Midline1 gene in secondary palate development by analyzing its expression and function in palatal shelf fusion and morphology. Initially, twenty mouse embryos were collected for each of the embryonic stages E13.5, E13.75, and E14.5. Whole-mount in situ hybridization was performed approximately ten times to optimize the experimental protocol and to analyze the expression pattern of Midline1 (MID1) in the palatal tissues at these developmental stages. Subsequently, palatal tissues from E13.5 embryos were treated with varying concentrations of Midline1 small interfering RNA (MID1 siRNA), and the knockdown efficiency was evaluated using Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR), with each concentration tested in triplicate. Based on the results, the most effective concentration, 100 nM MID1 siRNA, was selected for further experiments. Subsequently, twelve E13.5 palatal explants were allocated into two groups: six explants were treated with 100 nM MID1 siRNA (experimental group), and six with scrambled small interfering RNA(Scramble siRNA; control group). After 48 h of in vitro culture, hematoxylin and eosin (HE) staining was performed to evaluate the morphology of palatal shelf fusion. To evaluate apoptotic activity, Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL) staining was performed on both experimental and control groups. Finally, immunohistochemistry and Western blot analyses were conducted to examine the expression levels of Matrix Metalloproteinase 8 (MMP8) and Snail Family Transcription Factors (Snail) proteins in three biological replicates from each group. Midline1 deficiency resulted in incomplete palatal shelf fusion and significantly reduced apoptosis. Additionally, the knockdown of Midline1 led to the upregulation of Snail and MMP8 gene expression, indicating that Midline1 plays a critical role in regulating epithelial-to-mesenchymal transition and maintaining cytoskeletal stability during palate development. Midline1 is essential for normal secondary palate development. Its dysregulation disrupts palatal shelf fusion and morphology, potentially contributing to craniofacial abnormalities such as cleft palate. These findings provide new insights into the molecular mechanisms underlying palate development and suggest that Midline1 could be a therapeutic target for addressing cleft palate and related defects. 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

Endothelial-to-mesenchymal transition (EndMT) is a biological process that converts endothelial cells to mesenchymal cells with increased proliferative and migrative abilities. EndMT has been implicated in the development of pulmonary vascular remodeling in pulmonary arterial hypertension (PAH), a fatal and progressive lung vascular disease. Transforming growth factor β 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

Clear cell renal cell carcinoma (ccRCC), the most common subtype of kidney cancer, exhibits notable metabolic reprogramming. We previously reported elevated HDAC7, a class II histone deacetylase, in ccRCC. Here, we demonstrate that HDAC7 promotes aggressive phenotypes and in vivo tumor progression in RCC. HDAC7 suppresses the expression of genes mediating branched-chain amino acid (BCAA) catabolism. Notably, lower expression of BCAA catabolism genes is strongly associated with worsened survival in ccRCC. Suppression of BCAA catabolism promotes expression of SNAIL1, a central mediator of aggressive phenotypes including migration and invasion. HDAC7-mediated suppression of the BCAA catabolic program promotes Show less

The leading factor contributing to patient mortality is the local invasion and metastasis of tumors, which are influenced by the malignant progression of tumor cells. The epithelial-mesenchymal transition (EMT) is key to understanding malignancy development. EMT is a critical regulatory mechanism for differentiating cell populations initially observed during the neural crest and embryonic gastrulation formation. This process is closely associated with tumor metastasis in cancer and is also related to the maintenance of cancer stem cells. Flavonoids, known for their antioxidant properties, have been widely studied for their anticancer potential to protect plants from harmful environmental conditions. They have attracted considerable attention and have been the focus of numerous experimental and epidemiological studies to evaluate their potential in cancer treatment. In vitro and in vivo research has demonstrated that flavonoids can significantly impact cancer-related EMT. They may inhibit the EMT process by reducing the levels of Twist1, N-cadherin, ZEB1, integrins, SNAI1/2, CD44, MMPs, and vimentin while increasing E-cadherin levels and targeting the PI3K/AKT, NF-κB p65, and JAK2/STAT3 signaling pathways. In order to suppress the transcription of the E-cadherin promoter, several Zn-finger transcription factors, such as SNAI2, ZEB1, and ZEB2, and basic helix-loop-helix (bHLH) factors, such as Twist, may directly bind to its E-boxes. Overall, clinical cancer research should integrate the anticancer properties of flavonoids, which address all phases of carcinogenesis, including EMT, to improve the prospects for targeted cancer therapies in patients suffering from aggressive forms of tumors. Show less

Enhancer RNAs (eRNAs) are a pivotal class of enhancer-derived non-coding RNAs that drive gene expression. Here we identify the SNAI1 enhancer RNA (SNAI1e; SCREEM2) as a key activator of SNAI1 expression and a potent enforcer of transforming growth factor-β (TGF-β)/SMAD signaling in cancer cells. SNAI1e depletion impairs TGF-β-induced epithelial-mesenchymal transition (EMT), migration, in vivo extravasation, stemness, and chemotherapy resistance in breast cancer cells. SNAI1e functions as an eRNA to cis-regulate SNAI1 enhancer activity by binding to and strengthening the enrichment of the transcriptional co-activator bromodomain containing protein 4 (BRD4) at the local enhancer. SNAI1e selectively promotes the expression of SNAI1, which encodes the EMT transcription factor SNAI1. Furthermore, we reveal that SNAI1 interacts with and anchors the inhibitory SMAD7 in the nucleus, and thereby prevents TGF-β type I receptor (TβRI) polyubiquitination and proteasomal degradation. Our findings establish SNAI1e as a critical driver of SNAI1 expression and TGF-β-induced cell plasticity. 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

Limbal stem cell deficiency (LSCD) is a sight-threatening condition caused by the loss and/or dysfunction of limbal stem cells (LSCs), which are essential for corneal epithelial regeneration and homeostasis and are critical for maintaining corneal transparency. We have previously shown that specific inactivation of the endothelial mineralocorticoid receptor (MR) inhibits corneal neovascularization (CN) and that MR antagonists (MRA) improve corneal epithelial wound healing. This study investigated the therapeutic potential of MRA in LSCD and their mechanisms of action. Using a rat model of LSCD, systemic administration of spironolactone (SPL) or a more specific MRA, eplerenone, similarly reduced CN and corneal oedema, demonstrating MR-specific effects. SPL further limited inflammation, enhanced the corneal epithelial barrier, reduced corneal conjunctivalization and promoted nerve regeneration, highlighting its potential to improve corneal integrity. Transcriptomic analysis revealed that SPL upregulated genes associated with LSC maintenance (Tp63, Wnt6), corneal epithelial differentiation (Vdr, Fermt1, Ehf) and nerve regeneration (Sprr1a, Anxa1), while downregulating genes associated with angiogenesis (Kdr, Scube2), inflammation (Ccl2, Cxcl1) and fibrosis (Fbln1, Snai1). Conversely, transgenic rats overexpressing human NR3C2 encoding MR showed corneal epithelial irregularities and dysregulation of genes related to extracellular matrix remodeling and fibrosis (Matn3, Serpine2, Fmod, Bgn, Ddr2), angiogenesis (Nrp2, Scube1) and limbal cell function (Ifitm3). These findings demonstrate that activation of the MR pathway disrupts limbal and corneal homeostasis and that SPL effectively modulates critical mechanisms in LSCD, offering promising therapeutic potential to reduce CN and improve corneal epithelial barrier integrity. Show less

Snail1 transcriptional factor is essential for the epithelial to mesenchymal transition and for the acquisition by tumor cells of properties associated to this transition, such as increased invasion and chemoresistance. Snail1 function is mainly controlled post-translationally, through different modifications that directly or indirectly control Snail1 protein stability. In this review I describe these modifications, the enzymes that produce them and their relevance for Snail1 function, focusing particularly in polyubiquitination and phosphorylation. I also propose several explanations for the divergent effects of some of these modifications, since the phosphorylation of some residues have been reported to both promote and decrease Snail1 stability. Moreover, I discuss the possible causes of the observed Snail1 promiscuity in the interaction with the many factors involved in its regulation, on the basis of the Show less

Prefoldin1 (PFDN1), a molecular chaperone, is essential for stabilizing cytoskeletal proteins like actin and tubulin, supporting cellular processes such as survival, migration, and cell cycling. Recent evidence suggests that PFDN1 also influences key cancer-related signaling pathways. However, the complete mechanisms involved and the downstream genes implicated in such action remain relatively undiscovered. This study investigated the effects of PFDN1 silencing on cellular processes and gene expression in triple-negative breast cancer (TNBC) cells, focusing on its potential as a therapeutic target. MDA-MB-231 cells, a TNBC model, were transfected with PFDN1-targeting siRNA to knock down PFDN1 expression. The effects of PFDN1 silencing were assessed through various assays, including phase contrast and scanning electron microscopy (SEM) for morphological changes, colony formation and wound healing assays for proliferation and migration, and flow cytometry for cell cycle and apoptosis analysis. Gene expression changes were evaluated using a qRT-PCR array targeting 84 genes involved in cancer progression. PFDN1 silencing resulted in a 54.8% reduction in PFDN1 protein levels (p < 0.0001). Morphological analysis revealed cytoplasmic shrinkage, chromatin condensation, roughened membranes, and microvilli loss, consistent with apoptotic changes. Colony formation assays showed a 10.33% reduction in colony number and size (p < 0.05) in PFDN1-silenced cells. Migration was significantly impaired, with reduced wound closure observed in wound healing assays (p < 0.01). Flow cytometry revealed a G2/M phase arrest (p < 0.05) and increased early apoptotic populations (20.93% vs. 5.42% in controls, p < 0.01). Gene expression analysis showed downregulation of genes associated with angiogenesis (KDR, TEK), EMT (FOXC2, SNAI1), and hypoxia signaling (CA9, EPO), while proapoptotic genes, such as FASLG, were upregulated. This study highlights the critical role of PFDN1 in TNBC progression, demonstrating that its silencing disrupts survival, migration, cell cycling, and apoptosis pathways. PFDN1 knockdown also significantly alters the expression of key cancer-related genes, further impairing angiogenesis, EMT, and hypoxia adaptation. These findings suggest that targeting PFDN1 could be a promising therapeutic strategy for TNBC, warranting further investigation in preclinical models. Show less

Snail is a zinc finger transcription factor encoded by the SNAI1 gene and triggers a cellular process termed epithelial-mesenchymal transition (EMT) upon its increased expression and/or functional activation. Snail expression and activity are regulated by various extracellular stimuli, including cytokines and environmental factors. Transforming growth factor-β (TGF-β) is a Snail inducer that functions via Smad3-mediated transcriptional activation. In the present study, we identified a distal enhancer that modulates TGF-β-induced SNAI1 expression. ChIP-seq and Hi-C analyses showed that the enhancer is located 46 kb downstream of the SNAI1 gene; in TGF-β-stimulated cells, it associates with Smad3 and interacts with the SNAI1 proximal promoter. Inhibiting the activity of the enhancer using CRISPRi attenuated TGF-β-induced SNAI1 expression, stress fiber formation, and cell motility enhancement, suggesting that the enhancer mediates TGF-β-induced EMT. The enhancer contains a Smad-binding CAGA motif and an activator protein-1 (AP-1) binding motif that function in transcriptional activation. Ras-responsive element binding protein 1 (RREB1), a transcription factor required for TGF-β-induced Snail expression, regulated the basal activity of the enhancer but not its inducibility by TGF-β. In contrast to the enhancer, the association of Smad3 with the proximal promoter was not evident. These findings suggest that the proximal promoter and the distal enhancer respond to distinct signaling cues, integrate them, and cooperatively function to drive SNAI1 expression. Show less

This study aimed to investigate the effect of saltwater stir-fried Plantaginis Semen(SPS) on renal fibrosis in rats and decipher the underlying mechanism. Thirty-six Sprague-Dawley rats were randomly assigned into control, model, losartan potassium, and low-, medium-, and high-dose(15, 30, and 60 g·kg~(-1), respectively) SPS groups. Rats in other groups except the control group were subjected to unilateral ureteral obstruction(UUO) to induce renal fibrosis, and the modeling and gavage lasted for 14 days. After 14 consecutive days of treatment, the levels of serum creatinine(Scr) and blood urea nitrogen(BUN) in rats of each group were determined by an automatic biochemical analyzer. Hematoxylin-eosin(HE) and Masson staining were used to evaluate pathological changes in the renal tissue. Western blot and immunofluorescence assay were conducted to determine the protein levels of fibronectin(FN), collagen Ⅰ, vimentin, and α-smooth muscle actin(α-SMA) in the renal tissue. The mRNA levels of epithelial-mesenchymal transition(EMT)-associated transcription factors including twist family bHLH transcription factor 1(TWIST1), snail family transcriptional repressor 1(SNAI1), and zinc finger E-box binding homeobox 1(ZEB1), as well as inflammatory cytokines such as interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α), were determined by RT-qPCR. Human renal proximal tubular epithelial(HK2) cells exposed to transforming growth factor-β(TGF-β) for the modeling of renal fibrosis were used to investigate the inhibitory effect of SPS on EMT. Network pharmacology and Western blot were employed to explore the molecular mechanism of SPS in alleviating renal fibrosis. The results showed that SPS significantly reduced Scr and BUN levels and alleviated renal injury and collagen deposition in UUO rats. Moreover, SPS notably down-regulated the protein levels of FN, collagen Ⅰ, vimentin, and α-SMA as well as the mRNA levels of SNAI1, ZEB1, TWIST1, IL-1β, IL-6, and TNF-α in the kidneys of UUO rats and TGF-β-treated HK-2 cells. In addition, compared with Plantaginis Semen without stir-frying with saltwater, SPS showed increased content of specific compounds, which were mainly enriched in the mitogen-activated protein kinase(MAPK) signaling pathway. SPS significantly inhibited the phosphorylation of extracellular signal-regulated kinase(ERK) and p38 MAPK in the kidneys of UUO rats and TGF-β-treated HK2 cells. In conclusion, SPS can alleviate renal fibrosis by attenuating EMT through inhibition of the MAPK signaling pathway. Show less

Triple negative breast cancer (TNBC) is the most aggressive subtype and disproportionately affects African American women. The development of breast cancer is highly associated with interactions between tumor cells and the extracellular matrix (ECM), and recent research suggests that cellular components of the ECM vary between racial groups. This pilot study aimed to evaluate gene expression in TNBC samples from patients who identified as African American and Caucasian using traditional statistical methods and emerging Machine Learning (ML) approaches. ML enables the analysis of complex datasets and the extraction of useful information from small datasets. We selected four regions of interest from tumor biopsy samples and used laser microdissection to extract tissue for gene expression characterization via RT-qPCR. Both parametric and non-parametric statistical analyses identified genes differentially expressed between the two ethnic groups. Out of 40 genes analyzed, 4 were differentially expressed in the edge of tumor (ET) region and 8 in the ECM adjacent to the tumor (ECMT) region. In addition to statistical approach, ML was used to generate decision trees (DT) for a broader analysis of gene expression and ethnicity. Our DT models achieved 83.33 % accuracy and identified the most significant genes, including Show less