Hypoxia and nutrient deprivation are fundamental drivers of tumor aggressiveness and therapeutic resistance in hepatocellular carcinoma (HCC). While the involvement of neural components in the tumor microenvironment (TME) is increasingly recognized, the molecular transducers linking metabolic stress to neuron-tumor crosstalk remain elusive. Here, we identify ADAM23 (A disintegrin and metalloproteinase 23) as a hypoxia-responsive mediator that mediates communication between HCC cells and neuronal cells. ADAM23 expression was markedly upregulated in HCC cells under both chemical (CoCl2) and physical hypoxia (1% O2), a process further amplified by glucose deprivation and directly modulated by HIF-1α. Functional assays revealed that ADAM23 overexpression promotes epithelial-mesenchymal transition (EMT) and enhances cell viability under metabolic stress. Notably, sorafenib-resistant HCC cells (Huh7SR) exhibited high levels of ADAM23 secretion, which triggered proliferative and metabolic activation in neuronal SH-SY5Y cells. In 3D co-culture spheroid models, Huh7SR cells mixed with SH-SY5Y cells displayed significantly larger spheroid volumes and enhanced neuronal fluorescence compared with parental controls, suggesting that ADAM23-mediated interactions facilitate a supportive neural niche. Analysis of The Cancer Genome Atlas (TCGA) datasets and patient microarrays confirmed that ADAM23 is significantly overexpressed in HCC and positively correlates with HIF-1α expression. Moreover, elevated expression of ADAM23 was significantly correlated with poor overall survival. Collectively, our findings underscore ADAM23 as a critical metabolic-neural linker that promotes HCC progression and drug resistance. These findings suggest that the ADAM23-mediated neuron-tumor axis may represent a potential therapeutic target in aggressive HCC.
Lung cancer remains the leading cause of cancer-associated death, and aerobic glycolysis is a key hallmark of cancer. Ubiquitination is a post-translational modification involved in the regulation of lung cancer progression. The aim of this study was to investigate E3 ubiquitin ligase mouse double minute 2 (MDM2) and its ubiquitination on myosin heavy chain 11 (MYH11) in lung cancer progression and glycolytic reprogramming. Expression of MYH11 was analyzed using online databases. MDM2 and MYH11 mRNA and protein detection was performed by qPCR and Western blot. Cell proliferation, migration, and invasion were respectively detected using colony formation assay, wound healing assay, and transwell assay. Glycolysis was evaluated by measuring lactate production and glucose consumption. Epithelial-mesenchymal transition (EMT) and glycolysis-associated proteins were measured using Western blot. Xenograft tumor model was established for in vivo analysis. Co-immunoprecipitation (Co-IP) assay was performed to assess ubiquitination effect of MDM2 on MYH11 and their protein interaction. MYH11 was a down-regulated gene in lung cancer and it negatively correlated with glycolysis. Lung cancer cell proliferation, migration, invasion, EMT, and glycolysis were significantly inhibited following MYH11 overexpression. Tumor growth in vivo was also reduced by MYH11. Mechanistically, MDM2 mediated ubiquitination of MYH11 to induce MYH11 protein degradation and downregulation. Knockdown of MDM2 suppressed the malignant phenotypes of lung cancer cells via enhancing MYH11 protein stability in vitro and in vivo. These results suggest that MDM2 reduces MYH11 expression via ubiquitination-mediated degradation of MYH11 protein, thereby enhancing lung cancer cell proliferation, migration/invasion, EMT and glycolysis, and ultimately promoting tumor progression.
This study aims to investigate the biological function, molecular mechanisms, and impact on the tumor microenvironment of the ubiquitin-conjugating enzyme E2S (UBE2S) in the progression of kidney renal clear cell carcinoma (KIRC). Based on the TCGA and GEPIA databases, the relationship between UBE2S expression and patient prognosis was analyzed. Overexpression and knockdown models of UBE2S were established in Caki-1 and 786-O cell lines. Cell proliferation, apoptosis, and migration capabilities were assessed using CCK-8 assay, colony formation assay, flow cytometry, wound healing assay, and Transwell assay. Protein-protein interactions and expression regulatory mechanisms were validated by co-immunoprecipitation and Western blot. The proteasome inhibitor MG132 and ubiquitination assays were employed to investigate protein degradation mechanisms. Macrophage polarization was analyzed using a cell co-culture system, immunofluorescence staining, and Western blot. UBE2S is highly expressed in KIRC tissues and is associated with poor patient prognosis. Functional experiments demonstrate that UBE2S promotes KIRC cell proliferation and migration while inhibiting apoptosis. Mechanistically, UBE2S directly binds to and positively regulates CDC20 protein expression, and its oncogenic functions depend on CDC20. Furthermore, knockdown of UBE2S induces cuproptosis by activating FDX1 protein, thereby enhancing cellular sensitivity to cuproptosis inducers. UBE2S promotes FDX1 protein degradation via the ubiquitin-proteasome pathway. Additionally, tumor cells with high UBE2S expression promote macrophage polarization toward the M2 phenotype. UBE2S drives KIRC progression by regulating CDC20 expression and suppressing FDX1-mediated cuproptosis, thereby promoting tumor-associated macrophage polarization.
Previous genome-wide association studies have identified more than 400 prostate cancer (PCa) susceptibility loci. However, the tissue and spatial epithelial contexts through which inherited PCa risk may operate in the normal prostate remain incompletely understood. We analyzed normal human prostate spatial transcriptomic data using a gsMap-based framework to localize PCa association signals across spatially resolved prostate compartments. Spatial spots were clustered using SpaGCN and then manually curated and annotated based on matched histological images and canonical prostate marker gene expression. Because individual spatial spots may contain multiple cells, annotations were interpreted as compartment-enriched dominants rather than pure cell-type identities. PCa genetic association scores were aggregated within annotated compartments using Cauchy-combined p value and median spot-level p value as a complementary descriptive summary. Gene-level spatial correspondence was further assessed using Pearson correlation coefficients (PCCs) between gene-expression patterns and spot-level PCa gsMap signal. We annotated normal prostate tissue into four major compartments: basal epithelium-enriched, luminal epithelium-enriched, smooth muscle-enriched, and fibroblast stroma-enriched compartments. Basal epithelium-enriched compartments showed the strongest and most consistent enrichment for PCa association, with a Cauchy-combined p of 1.85 × 10-8 and a median spot-level based p of 3.67 × 10-7. Luminal epithelium-enriched compartments also showed strong enrichment, with a Cauchy p of 3.79 × 10-7 and a median-based p of 1.45 × 10-6. In contrast, fibroblast stroma-enriched compartments showed no evidence of enrichment (Cauchy p = 0.52; median p = 0.42), while smooth muscle-enriched compartments showed only modest evidence in the Cauchy-based analysis that was accompanied by a weaker median spot-level association signal (Cauchy p = 8.95 × 10-4; median p = 0.18). PCC analysis showed that genes with the strongest spatial correspondence to PCa gsMap signal were predominantly epithelial-associated, including CDH1, VAMP8, GMNN, TSPAN1, and FOXA1. In normal human prostate tissue, inherited PCa association localizes preferentially to basal epithelium-enriched and luminal epithelial-enriched compartments rather than to smooth muscle-enriched or fibroblast stroma-enriched compartments. Gene-level PCC patterns further support spatial alignment between PCa-associated gsMap signal and epithelial transcriptional programs. These findings provide proof-of-concept evidence for the spatial localization of inherited PCa susceptibility through the integration of large-scale population-based genetic data and spatial transcriptomics.
Melanoma is a highly aggressive skin cancer with poor prognosis, often linked to excessive UV exposure. Despite advancements in immunotherapy and targeted treatments, melanoma's invasiveness and metastatic potential remain significant challenges. This study aimed to identify novel molecular targets associated with melanoma progression. Consensus clustering analysis revealed three melanoma subtypes, with the C3 subtype exhibiting the worst prognosis. Weighted gene co-expression network analysis (WGCNA) identified FOXQ1 as a key driver in this subtype. FOXQ1 expression was elevated in clinical melanoma tissues and cell lines, and silencing FOXQ1 in vitro reduced melanoma cell proliferation, migration, and induced apoptosis. To explore its downstream targets, FOXQ1 was found to repress the expression of SSBP2, a gene inversely correlated with FOXQ1. Functional assays demonstrated that silencing SSBP2 promoted melanoma malignancy. Rescue experiments showed that knockdown of SSBP2 reversed the tumor-suppressive effects of FOXQ1 silencing. These results suggest that the FOXQ1/SSBP2 regulatory axis plays a critical role in melanoma progression and may serve as a potential therapeutic target to improve melanoma treatment outcomes.
Cellular senescence plays a critical role in physiological and pathological processes. This study aims to elucidate the contribution of cellular senescence-related genes to disease etiology. We investigated a cohort study of 439,501 individuals, which included 22 cancers and 9 non-cancer diseases. We found that HLA-E and HLA-G-associated senescence in epithelial and immune cells were specific oncogenic factors for prostate and lung cancers. MAP2K4 was implicated as a risk factor for breast cancer, while ZFP36L1 and STAT3 were associated with a reduced risk of inflammatory bowel disease (IBD). Notably, ETS2-mediated inhibition of the senescence-associated secretory phenotype (SASP) was associated with decreased disease risk. Furthermore, single-cell level analysis confirmed that the dynamics of these marked gene expressions in immune cells was related to reduced disease risk, while upregulation in epithelial cells correlated with increased disease risk. In parallel, co-localization analyses corroborated these associations, explaining potential regulatory mechanisms underlying disease risk variants. These findings enhance our understanding of how cellular senescence works on disease susceptibility and provide potential targets for therapeutic interventions and precision medicine approaches.
The family with sequence similarity 3 (FAM3) family has four members (FAM3A, FAM3B, FAM3C, and FAM3D), which have been proven to contribute to tumorigenesis. However, their correlations with ovarian cancer (OV) prognosis and regulatory functions in OV are poorly understood. Here, the expression of four FAM3 family members in OV was analyzed, and functional assays were conducted to explore the effects and underlying mechanisms of FAM3C in OC progression and chemical resistance. FAM3C was upregulated in OV tissues, and higher FAM3C was associated with poorer overall survival and shorter progression-free survival in OV patients. Upregulating FAM3C promoted OV cell proliferation and growth and reduced DNA damage and sensitivity to olaparib. In contrast, downregulating FAM3C restrained OV cell proliferation, promoted DNA damage, and enhanced olaparib sensitivity. Mechanistically, FAM3C overexpression promoted p-STAT3 and HIF-1α expression, which was suppressed after treatment with the HIF-1α inhibitor LW6 or the STAT3 inhibitor static. In vivo assay confirmed that inhibiting STAT3 reversed the FAM3C-mediated promotive effects on tumor growth and enhanced the sensitivity of A2780 cells to olaparib. Taken together, our findings demonstrate that FAM3C promotes OV development and reduces olaparib sensitivity by activating the HIF-1α/STAT3 pathway. FAM3C could serve as a potential diagnostic marker and anticancer target in OV.
Cervical cancer (CC) is one of the most common gynecological malignancies. Although E74-like factor 3 (ELF3) has been implicated as an oncogenic driver in multiple cancers, its expression pattern and functional role in CC remain unclear. An integrated analysis of transcriptome, proteome, and transcription-factor libraries was performed to identify the most significantly upregulated gene in CC, and GEPIA3.0 was used to confirm ELF3 overexpression. The expression levels of ELF3, guanosine monophosphate synthetase (GMPS), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4), as well as the knockdown efficiencies of ELF3 and GMPS, were evaluated by RT-qPCR and Western blot. The effects of ELF3 or GMPS silencing on proliferation, migration, apoptosis, and ferroptosis were assessed by colony-formation, Transwell, and flow cytometry assays. Direct targeting of GMPS by ELF3 was verified by chromatin immunoprecipitation and dual-luciferase reporter assays, and a xenograft model in nude mice was established to demonstrate that ELF3 depletion suppresses CC growth. ELF3 was identified as the most significantly upregulated gene in CC. Silencing either ELF3 or GMPS inhibited proliferation and migration while inducing ferroptosis. Mechanistically, ELF3 transcriptionally regulated GMPS, and GMPS overexpression fully rescued the effects of ELF3 knockdown. In vivo, ELF3 depletion markedly restrained CC xenograft growth. These findings demonstrate that ELF3 functions as an oncogenic transcription factor in CC by upregulating GMPS, thereby enhancing proliferation and migration and restraining ferroptosis. Targeting the ELF3/GMPS axis represents a promising therapeutic strategy for CC.
In colorectal cancer (CRC), metastasis drives fatalities, necessitating the discovery of therapeutic targets modulating its progression. Though the secreted protein-coding gene leucine-rich repeat neuronal 4 (LRRN4) was shown to be highly overexpressed in CRC and linked to decreased survival in the analysis of TCGA-CRC datasets, little is known about its function as well as mechanism in CRC metastasis. Thus, our work aimed to investigate LRRN4 functions and its role in CRC metastasis. We first confirmed that CRC metastasis was related to raised LRRN4 expression in CRC tissues. LRRN4 depletion using siRNA/shRNA inhibited CRC metastasis in vitro and in vivo, decreasing vimentin and increasing E-cadherin levels, whereas its overexpression yielded opposite effects. Mechanistically, LRRN4 promoted CRC metastasis by upregulating IL23A expression and triggering JAK2-STAT3 signaling, as evidenced by LRRN4 depletion-reduced IL23A expression and downstream JAK2-STAT3 activity; the STAT3 inhibitor and recombinant human IL23 protein rescued the promotion and inhibition of cell migration induced by LRRN4 overexpression and knockdown, respectively. Kaplan-Meier analysis and the K-independent samples nonparametric test revealed that the upregulated marker proportion was positively related to the likelihood of metastases, and people having raised IL23A and LRRN4 levels had the lowest survival. Finally, elevated LRRN4 expression in CRC tissues was correlated with promoter hypomethylation. Thus, LRRN4 is upregulated by promoter DNA hypomethylation, and the LRRN4-IL23A-JAK2-STAT3 axis is a critical driver of CRC metastasis, representing a promising therapeutic target and prognostic biomarker.
Liver hepatocellular carcinoma (LIHC) is a molecularly heterogeneous malignancy for which additional genetically supported biomarkers and functional regulators remain to be identified. We aimed to identify candidate genes through integrative genetic and transcriptomic screening, then determine the cellular context and functional relevance of the leading candidate. We applied Summary-data-based Mendelian Randomization to FinnGen LIHC genome-wide association summary statistics and GTEx v8 liver cis-eQTL summary data. A 1000 Genomes European-ancestry panel provided the linkage disequilibrium reference. Candidate genes were cross-referenced with Gene Expression Omnibus differentially expressed genes and TCGA-LIHC prognostic genes. After ATP13A2 emerged from this screen, we used bulk, single-cell, and spatial transcriptomic analyses to characterize its expression and cellular context. HUVEC knockdown, migration, tube formation, qRT-PCR, and Western blotting were then used for functional assessment. ATP13A2 was the only gene shared by the SMR-prioritized, GEO differential-expression, and TCGA-LIHC prognostic sets. ATP13A2 expression was higher in LIHC tissues and independently associated with worse survival in the TCGA-LIHC cohort. Single-cell analysis subsequently localized ATP13A2 predominantly to tumor endothelial cells, and spatial and pathway analyses associated ATP13A2-positive endothelial cells with angiogenic programs and inferred PTN-NCL signaling. In HUVECs, ATP13A2 knockdown reduced migration and tube formation and was accompanied by reduced ERK1/2 and p38 MAPK phosphorylation. An unbiased integrative screen prioritized ATP13A2 as a genetically supported LIHC candidate, after which single-cell analysis identified its tumor-endothelial context. Functional knockdown data support an association with endothelial angiogenic phenotypes. Rescue, in vivo, and independent clinical validation remain required.
Herein, using the LSL-KrasG12D/+; p48Cre/+ transgenic mouse model [that recapitulates the full spectrum of human disease, progressing from early pancreatic intraepithelial neoplasia lesions to invasive pancreatic ductal adenocarcinoma (PDAC)], we determined the chemopreventive effect of long-term (chronic) administration of Bitter Melon Juice (BMJ-standardized lyophilized juice processed from the fruits of Momordica charantia) against pancreatic cancer. Starting at 6 weeks of age, female LSL-KrasG12D/+; p48Cre/+ mice were subjected to BMJ intervention (oral gavage, dose: 200 mg/kg body wt., 5 days/week) for 35 weeks (study end: 41 weeks of age), and pathological and molecular changes in the pancreas were assessed. Results indicated that BMJ shifted the pancreatic tumor profile from moderately differentiated adenocarcinoma in controls towards a more heterogeneous profile enriched for well-differentiated lesions indicating that prolonged exposure to BMJ bioactives results in a less aggressive phenotype. BMJ significantly reduced proliferative index (~24%) and induced a ~ 4-fold increase in apoptosis in the pancreas. The effects on AMPK activation (~2-fold increase) and decrease in the expression of [glucose (GLUT-1) and lactate (MCT-4) transporter levels (∼49%-69%,), NF-κB/p65 (~40%), EMT marker-Vimentin (~46%), fibrotic microenvironment marker (α-SMA, ~29%) and Mucin-1 (MUC-1, ~45%)] indicate that BMJ modulates several interconnected pathways essential for PDAC growth and progression. Plasma proteomic profiling indicated that Fractalkine (CX3CL1), MIP-3β (CCL19), MIP-3α (CCL20), and SCF were decreased by BMJ, while Axl and TNF-alpha were overexpressed in BMJ-treated animals compared to positive controls. These results encourage further research into bitter melon compounds as a potential chemopreventive option for pancreatic cancer, especially for high-risk groups.
This study investigated the mechanistic role of DUSP9 in hepatoblastoma (HB) development and progression to provide new insights into the diagnosis and treatment of pediatric HB. Pediatric HB mRNA expression data were obtained from the GEO database. Differentially expressed genes (DEGs) between patients with metastatic and non-metastatic HB were analyzed using the R software, followed by a functional annotation of the DEGs. Least Absolute Shrinkage and Selection Operator regression and Support Vector Machine-Recursive Feature Elimination machine learning methods were employed to further screen for DEGs and identify DUSP9 as a key feature gene. The diagnostic efficacy of DUSP9 was evaluated using the area under the receiver operating characteristic curve. Additionally, in vitro and in vivo functional experiments were conducted to determine the effects of DUSP9 on HB progression. A total of 33 DEGs were identified. Gene Ontology analysis revealed that these DEGs were mainly associated with xenobiotics and fatty acid metabolism. Kyoto Encyclopedia of Genes and Genomes analysis indicated involvement in regulating active transmembrane transporter activity and cytochrome P450-mediated xenobiotic metabolism. Machine learning methods identified DUSP9 as a pivotal feature gene that was significantly upregulated in HB tumor tissues. In vitro functional assays demonstrated that DUSP9 knockdown markedly inhibited the proliferation and migration of HB cells. Additionally, western blot analysis revealed increased expression of epithelial markers and decreased expression of mesenchymal markers, along with a significant reduction in the phosphorylation of markers related to the MAPK signaling pathway. DUSP9 plays a crucial role in HB cell proliferation, migration, and epithelial-mesenchymal transition (EMT), potentially promoting tumor progression by regulating the MAPK signaling pathway.
Glioblastoma (GBM), a Grade IV malignant brain tumor, accounts for over half of all gliomas and remains resistant to current therapies, requiring the development of novel treatment strategies. Aberrant Ras signaling and PI3K/Akt hyperactivation and RAF/MEK/ERK pathways are major oncogenic drivers, implicated in approximately 30% of cancers, including GBM. Chromenes, particularly 4H-chromenes, possess diverse anticancer activities, but their application in GBM remains limited. In this study, we evaluated the efficacy of a novel chromene derivative, ethyl 4-(3,5-dichloro-2-hydroxyphenyl)-5,7-dihydroxy-2-methyl-4H-chromene-3-carboxylate (4H-CRCXL), using a Drosophila melanogaster model of RasV12-driven gliomagenesis. In silico docking revealed strong interactions of 4H-CRCXL with Ras, p-Akt, p-ERK, and Bcl-2, suggesting multi-targeted inhibition. In vivo administration of 4H-CRCXL resulted in significant phenotypic rescue, improved CNS morphology, enhanced survival, and attenuated RAS-driven tumor phenotypes. The compound suppressed glial hyperproliferation, reduced oxidative stress, restored metabolic homeostasis, and induced apoptosis, demonstrating a broad spectrum of anticancer effects. These findings highlight 4H-CRCXL as a promising lead compound for targeting RAS-driven gliomagenesis and support its further evaluation in mammalian glioma models.
CXCL13+ T cells and LAMP3+ dendritic cells (DCs) are pivotal players in orchestrating anti-tumor immune responses, particularly within tumor tertiary lymphoid structures (TLS). However, their heterogeneity, differentiation trajectories, and clinical relevance in bladder cancer remain incompletely defined. This study integrated single-cell RNA sequencing (scRNA-seq) data (16 bladder cancer patients, 113,905 post-quality-control cells) and spatial transcriptomics to characterize CXCL13+ T cell/LAMP3+ DC subsets, their differentiation pathways (via Velocyto trajectory analysis), and intercellular crosstalk (via receptor-ligand mapping). A risk model (DTscore) was constructed using marker genes of these cells and validated in the IMvigor210 (atezolizumab-treated bladder cancer) and TCGA-BLCA cohorts. scRNA-seq clustering identified 10 immune and 3 nonimmune cell types, with T cells stratified into 8 subpopulations (including CD4+CXCL13+ T cells and CD8+CXCL13+ T cells) and DCs into 9 subgroups (including LAMP3+ DCs). Receptor-ligand mapping and spatial transcriptomics confirmed functional crosstalk between CXCL13+ T cells and LAMP3+ DCs via key pairs (e.g., CCR7-CCL19, CXCR5-CXCL13, PDCD1-CD274) within TLS. The DTscore was developed using 8 marker genes (TSHZ2, ALOX5AP, GADD45G, TXN, CHN1, CCL19, CXCL13, ICA1) and exhibited robust prognostic and predictive performance: In the IMvigor210 cohort, high DTscore correlated with significantly poorer overall survival (OS) and a 3.27-fold lower immunotherapy response rate (11% vs. 36%, p = 4.23e-07); multivariate Cox regression confirmed DTscore as an independent OS predictor (hazard ratio = 1.97, p < 0.001). DTscore retained prognostic value in TCGA-BLCA (OS: p = 0.003; disease-specific survival: p < 0.001) and effectively predicted atezolizumab response even in the "immune desert" phenotype (p = 0.04). Combining DTscore with tumor mutational burden/tumor neoantigen burden yielded an AUC of 0.8122 for response prediction. Additionally, high DTscore was associated with higher OS hazard ratios in patients with wild-type TTN, RB1, EP300, or FGFR3 (all p < 0.01), while FGFR3 mutations correlated with lower immune checkpoint/CXCL13 expression. This study delineates the heterogeneity and interactions of CXCL13+ T cell/LAMP3+ DC subsets in bladder cancer TLS and validates DTscore as a robust tool for predicting OS and immunotherapy response, offering a potential guide for personalized bladder cancer treatment.
Oral squamous cell carcinoma (OSCC) exhibits high recurrence rates, and immune escape within the tumor microenvironment is a critical barrier to effective therapy. This study investigates whether cancer-associated fibroblast (CAF)-derived FGF7 promotes immune escape in OSCC through JAK/STAT3 pathway-mediated PD-L1. Tumor and adjacent tissues from OSCC patients were analyzed for FGFF7 expression, and the correlations between FGF7 and clinicopathological features were analyzed. CAFs were isolated from OSCC, transfected with plasmids targeting FGF7, and assessed for activation markers, 3D spheroid formation, and epithelial-mesenchymal transition (EMT) proteins. Co-culture systems were established to evaluate the malignant behaviors of OSCC cells exposed to modified CAFs through proliferation, migration, invasion, and apoptosis assays. OSCC cells were co-cultured with CD8+ T cells, followed by measurement of JAK/STAT3 phosphorylation, PD-L1 expression, T-cell activation markers, and immune-related genes. In vivo effects were examined in xenograft models. FGF7 was overexpressed in OSCC tissues and cells, primarily originating from CAFs. FGF7 knockdown in CAFs suppressed activation markers, EMT, and malignant behaviors of OSCC cells. Mechanistically, FGF7 knockdown reduced JAK/STAT3 phosphorylation and PD-L1 expression, impairing T-cell cytotoxicity and antigen presentation. A specific STAT3 inhibitor completely reversed FGF7-induced PD-L1 upregulation. ChIP-qPCR confirmed that STAT3 directly binds to the PD-L1 promoter and activates its transcription. In vivo, CAF-specific FGF7 knockdown attenuated tumor growth and collagen deposition while increasing CD8+ T-cell infiltration and apoptosis. Combined FGF7 knockdown and anti-PD-L1 therapy synergistically enhanced these effects. CAF-derived FGF7 drives immune escape and OSCC progression by upregulating PD-L1 via JAK/STAT3 signaling.
Overactivation of thioredoxin reductase 1 (TrxR1) is implicated in tumor development. This study aims to elucidate the efficacy and mechanisms by which neferine (Nef) exerts its anti-tumor effects in pancreatic cancer through targeting TrxR1. The results demonstrate that Nef directly binds to and inhibits the enzymatic activity of TrxR1, leading to an abnormal accumulation of reactive oxygen species (ROS) in mitochondria. This accumulation subsequently results in a decrease in mitochondrial membrane potential, DNA damage, and the activation of apoptotic pathways, characterized by changes in the Bax/Bcl-2 ratio and a cascade reaction involving caspases. Reversal experiments indicated that the ROS scavenger NAC significantly counteracted the inhibitory effects of Nef on pancreatic cancer cell proliferation and its induction of apoptosis. Genetic experiments confirmed that the knockdown of TrxR1 enhances the anti-pancreatic cancer effects of Nef. Furthermore, a nude mouse xenograft tumor model validated the in vivo efficacy of Nef in suppressing tumor growth via TrxR1 inhibition. Bioinformatic analysis suggested that elevated TrxR1 expression is associated with a poor prognosis in pancreatic cancer. Collectively, our results confirm that Nef is a novel inhibitor of TrxR1 and provide experimental evidence for a therapeutic strategy targeting TrxR1.
Gastric cancer (GC) remains a leading cause of cancer-related mortality worldwide, with limited therapeutic advancements despite progress in early detection. Serine hydroxymethyltransferase 2 (SHMT2), a key metabolic enzyme, and fructose-1,6-bisphosphate aldolase A (ALDOA), a glycolytic enzyme, are implicated in tumor progression. However, the molecular mechanisms linking SHMT2 and ALDOA in GC remain unclear. This study investigates how SHMT2 regulates ALDOA expression via m6A RNA modification to drive GC malignancy. Bioinformatic analyses (TCGA, LinkedOmics, and SRAMP) were used to assess SHMT2 expression in GC patients and identify its correlated genes. In vitro experiments (CCK-8, EdU, Transwell, and wound healing) evaluated the effects of SHMT2 overexpression or knockdown on GC cell proliferation, migration, invasion, and glycolysis. m6A modification of ALDOA was analyzed via MeRIP-PCR and dual-luciferase assays, while RNA stability was assessed using actinomycin D treatment. Xenograft models validated SHMT2's role in vivo. SHMT2 was upregulated in GC tissues and cell lines, correlating with advanced tumor stages and poor prognosis. SHMT2 knockdown suppressed GC cell viability, migration, invasion, and glycolysis, while overexpression enhanced these traits. Mechanistically, SHMT2 increased S-adenosylmethionine levels, promoting ALDOA m6A modification, likely mediated through the predicted site 1 (position 291). This modification stabilized ALDOA mRNA via IGF2BP1 recognition, an m6A reader. ALDOA overexpression reversed the tumor-suppressive effects of SHMT2 knockdown. In vivo, SHMT2 depletion reduced tumor growth and Ki67 expression in xenograft models. In conclusion, SHMT2 drives GC progression by enhancing ALDOA expression through m6A modification and IGF2BP1-mediated stabilization. Targeting the SHMT2-ALDOA axis represents a promising therapeutic strategy for gastric cancer.
Neutrophil extracellular traps (NETs) play crucial roles in cancer progression, but their regulatory mechanisms in breast cancer remain poorly understood. We developed a NETs-related prognostic risk model using TCGA breast cancer data and identified key biomarkers through bioinformatics analysis. AZU1 expression was validated in clinical samples using qRT-PCR and immunohistochemistry. In vitro experiments investigated AZU1's effects on neutrophil activation and NET formation using recombinant protein treatment, co-culture assays, and flow cytometry. Mechanistic studies employed phospholipase C (PLC) inhibition and PAD4 knockdown approaches. An orthotopic mouse model validated in vivo findings. Four NETs-related genes (F2RL2, AZU1, IL33, ELANE) constituted a robust prognostic model with good predictive performance. AZU1 showed significant upregulation in breast cancer tissues and correlated with advanced tumor stages. AZU1 overexpression in breast cancer cells enhanced neutrophil recruitment and NET formation through PLC signaling activation. Recombinant AZU1 dose-dependently activated neutrophils, promoted NET formation, and enhanced cancer cell invasion via epithelial-mesenchymal transition induction. PLC inhibition and PAD4 knockdown effectively blocked AZU1-induced neutrophil activation. In vivo experiments confirmed that AZU1 overexpression accelerated tumor growth and metastasis, while PAD4 inhibition reversed these effects. AZU1 promotes breast cancer progression through PAD4-dependent NET formation, representing a potential therapeutic target for breast cancer treatment.
Sodium overload has recently emerged as a critical metabolic stressor involved in cancer progression; however, its molecular characteristics and clinical relevance in acute myeloid leukemia (AML) remain unexplored. RNA-seq data sets, clinical annotations, and mutational profiles of AML patients were annotations from The Cancer Genome Atlas and integrated with Genotype-Tissue Expression normal samples. Sodium overload-related genes (SORGs) were obtained from GeneCards. Differentially expressed SORGs (DESORGs) screened by applying the limma statistical model, followed by univariate Cox proportional hazards regression, consensus clustering, functional enrichment, immune infiltration analysis, and pathway evaluation. A prognostic signature was developed through least absolute shrinkage and selection operator regression followed by multivariate Cox modeling. The model's performance was further verified in two external GEO data sets (GSE71014 and GSE37642). Nomogram construction, subgroup analysis, tumor mutational burden (TMB) assessment, drug sensitivity prediction, transcription factor (TF) analysis, and competing endogenous RNA (ceRNA) network analyses were also performed. A total of 57 DESORGs were identified, and 2 sodium overload-related molecular subtypes exhibited distinct survival, immune infiltration, and inflammatory pathway activation. A robust four-gene signature (DOCK1, GABRE, HTR7, ACSM1) stratified patients into high- and low-risk categories with significantly different survival across training and validation cohorts. High-risk patients displayed increased immune infiltration, higher TMB, reduced sensitivity to multiple chemotherapeutic drugs, and inferior predicted response to PD-L1 blockade. TF and ceRNA networks revealed multilayered transcriptional and post-transcriptional regulation of the signature genes. This study identifies sodium overload-related molecular heterogeneity in AML and establishes a validated four-gene prognostic signature that integrates genomic, immunologic, and therapeutic features, offering potential utility for personalized risk assessment and treatment optimization.
ABSTRACT As a key regulator of N 6 ‐methyladenosine (m 6 A) modification, methyltransferase‐like 14 (METTL14) has been implicated in the progression of various cancers; however, its functional role in breast cancer remains controversial. Isorhapontigenin (ISO), a natural polyphenolic compound, has been identified as a METTL14 agonist with antitumor potential in multiple malignancies. Nevertheless, the biological function of ISO in breast cancer, particularly its mechanism of regulating METTL14 and downstream signaling pathways, has not been fully elucidated. This study aimed to explore the effects of ISO on breast cancer cell viability, colony‐forming efficiency, and ferroptosis and investigate whether ISO exerts these effects by regulating METTL14 expression. Bioinformatics analyses were performed to identify differentially expressed m 6 A‐related genes in breast cancer tissues. CCK‐8 and colony formation assays were used to evaluate the effects of ISO on breast cancer cell viability and colony‐forming efficiency. Ferroptosis was assessed by quantifying ferroptosis‐related indicators, including reactive oxygen species, glutathione, malondialdehyde, and intracellular Fe 2+ levels. METTL14‐overexpressing cell lines, as well as METTL14‐ and SLC7A11‐silenced cell lines, were constructed to explore gene functions. Bioinformatics analysis revealed that METTL14 is downregulated in basal‐like and HER2‐enriched breast cancer subtypes, and METTL14 overexpression suppressed cell viability and colony‐forming efficiency and promoted ferroptosis in these METTL14‐low subtypes. ISO also suppressed cell viability and colony‐forming efficiency and induced ferroptosis in these cell subtypes. Mechanistically, ISO exerted its effects by upregulating METTL14 expression, which in turn induced m 6 A modification of SLC7A11 mRNA. In conclusion, ISO reduced cell viability and colony‐forming efficiency, and promoted ferroptosis in HER2‐enriched and basal‐like breast cancer cells through promoting METTL14‐dependent m 6 A modification of SLC7A11 mRNA. These findings suggest that ISO may serve as a candidate therapeutic agent for the treatment of HER2‐enriched and basal‐like breast cancer.
