Background Thyroid cancer (TC) has traditionally been regarded as an indolent malignancy. However, it retains the potential for metastasis and lethality, underscoring its status as a malignant tumor. To explore the interplay between serum metabolite profiles, gut microbial communities, and TC metastasis, we conducted a comprehensive study. Methods Serum metabolites were analyzed using untargeted metabolomics methods to characterize the metabolic changes. Two diet-derived metabolites, creatine (Cr) and α-linolenic acid (ALA), were selected for in vitro validation. Finally, 16S rRNA gene sequencing was used to characterize the intestinal microbial composition and analyze the correlation between differential metabolites and differential intestinal microbiota. Results LC-MS analysis of serum metabolites identified Arginine and proline metabolism and Arginine biosynthesis as the primary metabolic pathways altered in TC metastasis. Both Cr and ALA effectively inhibited EMT, migration, and invasion of TC cell lines, TPC1 and FTC-133. The analysis of gut microbiota composition revealed that Cr and ALA showed significant correlations with the bacteria genera, Porphyromonas and Papillibacter. Conclusions This study highlights that TC metastasis is associated with significant alterations in the serum metabolite profiles of patients, and Cr and ALA play a role in inhibiting TC development. Furthermore, Cr and ALA were correlated with Porphyromonas and Papillibacter, implying their potential link with food-borne components and their connected impact on TC metastasis. The findings provide new insights into the role of metabolites and gut microbiota in TC progression, suggesting novel avenues for dietary intervention in cancer treatment.
BACKGROUND:Tissue-based immunohistochemistry (IHC) for programmed death-ligand 1 (PD-L1) remains the clinical standard for guiding immunotherapy selection in non-small cell lung cancer (NSCLC) but is constrained by intratumoral spatial heterogeneity and procedural sampling risks. Circulating soluble PD-1 (sPD-1) and soluble PD-L1 (sPD-L1) generated either through alternative mRNA splicing or by ADAM10/17-mediated proteolytic shedding offer minimally invasive liquid-biopsy alternatives that capture systemic immune dynamics in real time. METHODS:This review synthesizes clinical and translational data from prospective validation cohorts, surgical trials, and multi-center meta-analyses to evaluate the stage-stratified diagnostic utility, prognostic reliability, and treatment-response predictive value of sPD-1 and sPD-L1 across diverse therapeutic contexts. RESULTS:Cross-sectional baseline assessments show modest standalone diagnostic sensitivity due to significant absolute concentration overlaps between oncological cohorts and healthy controls. However, elevated pre-treatment sPD-L1 functions as a robust independent negative prognostic indicator of overall survival (OS) and a predictor of primary resistance to immune checkpoint inhibitors (ICIs) across advanced stages, while demonstrating no prognostic relevance under conventional cytotoxic chemotherapy. Longitudinally, an acute postoperative sPD-L1 increase of ≥20% at four months marks micro-metastatic persistence and predicts disease recurrence in early-stage surgical cohorts (Odds Ratio = 10.29). Conversely, sPD-1 demonstrates distinct stage-specific volatility; pre-treatment profiles lack stable prognostic indicators, but an early on-treatment plasma surge during anti-PD-1 monotherapy independently maps to prolonged progression-free survival and superior OS (Hazard Ratio = 0.24). Finally, composite liquid biosignatures (e.g., sCombo or joint bsPD-L1/MMP screening) significantly optimize predictive resolution over single-analyte measurements. CONCLUSIONS:Circulating sPD-L1 serves as a reliable negative indicator of therapeutic efficacy and surgical durability, whereas on-treatment tracking of sPD-1 captures protective host T-cell clonal reactivation. Cross-platform assay standardization and predefined cutoff validation remain essential thresholds before these liquid biomarkers can be implemented into standard clinical practice and thoracic surgery. Integrating these circulating checkpoints into high-throughput multi-omics platforms and AI-based analytics is a promising next step toward reproducible predictive signatures that could guide personalized treatment in thoracic oncology.
Circulating tumor DNA (ctDNA) is characterized by low abundance and fragmentation, limiting the development of genetic variant detection technologies. In this study, we established a highly sensitive and specific assay by combining peptide nucleic acid (PNA)-mediated PCR clamping with CRISPR/Cas13a trans-cleavage detection. A PNA probe targeting the wild-type (WT) EGFR T790M allele was designed to suppress WT amplification during PCR, while minimally affecting mutant allele amplification. By combining the target specificity of Cas13a for mutant alleles with the WT-suppression capability of PNA-PCR, we achieved a dual-enrichment effect for mutant detection. When applied to EGFR T790M mutation detection, the assay reached an analytical sensitivity of 0.02%. We established a standard curve for T790M detection using cell-free DNA standards. Clinical validation in 20 plasma samples from lung adenocarcinoma patients demonstrated that the PNA-Cas13a assay achieved a diagnostic sensitivity of 93.3% (95% CI: 68.1%-99.8%) and a specificity of 100% (95% CI: 47.8%-100%), with detection concordance comparable to or improved over ARMS-PCR in this pilot cohort. The results suggest its preliminary diagnostic utility in liquid biopsy. In conclusion, the PNA-Cas13a assay enables sensitive and specific detection of EGFR T790M mutations in ctDNA, is readily adaptable to multiple gene loci, and holds promise for clinical monitoring of tumor drug resistance.
BACKGROUND:Systemic lupus erythematosus (SLE) is a highly complex autoimmune disorder that poses considerable treatment challenges. Among them, cutaneous lupus erythematosus (CLE) is one of the most common and earliest clinical symptoms. Dihydroartemisinin (DHA) is a semi-synthetic derivative of artemisinin, which is extracted from the traditional Chinese herb Artemisia annua. Recent studies have suggested that DHA has immunosuppressive effects. This study aims to further explore the role of DHA in SLE-related skin conditions and its effects on keratinocytes (KC). METHODS:A mouse model of SLE was created by inducing the disease with Pristane, and skin damage was assessed using the CLASI scoring system. Histological alterations were analyzed through H&E and Masson staining. IF was used to detect IgG and C3 deposits in the skin tissue, while immunohistochemistry evaluated the expression of K16. An in vitro SLE skin model was developed by exposing mouse keratinocytes to ultraviolet light. Subsequently, cell proliferation was measured using the EDU assay, ROS levels were determined by DCFH-DA fluorescence, cytokine levels were quantified via ELISA, and cell chemotaxis was assessed using a Transwell assay. Previous research has shown that the MAPK signaling pathway plays a role in treating SLE with prednisone and DHA, and further MCODE analysis identified MAPK14 as the key gene within this pathway. RESULTS:The findings demonstrated that combining Prednisone (PDN) with DHA can reduce skin lesions associated with SLE and the abnormal activation of KC in mice. Mechanistically, we discovered that PDN and DHA bind together to suppress inflammatory activation of KC by targeting MAPK14, thereby modulating the Treg/Th17 balance. CONCLUSION:This research highlights the protective role of DHA in SLE-related skin inflammation and offers experimental support for its ability to improve abnormal KC activation via immune regulatory pathways.
BACKGROUND:The DExD/H-box (DDX) helicase family plays critical roles in RNA metabolism and has been implicated in tumorigenesis. However, the pan-cancer activities and prognostic potential of DDX52, especially in liver hepatocellular carcinoma (LIHC), are largely unknown. METHODS:We comprehensively analyzed DDX52 across pan-cancer using TCGA, GEO, HPA, and SpatialTME databases, assessing its differential expression, prognosis, genetic alterations, Tumor Mutational Burden (TMB), Microsatellite Instability (MSI), and immune microenvironment features. In LIHC, we performed functional enrichment, PPI network, single-cell sequencing, and drug sensitivity analyses. Molecular docking explore potential small-molecule inhibitors of DDX52. Finally, we validated DDX52 expression and function in LIHC. RESULTS:DDX52 mRNA and protein expression were significantly upregulated in multiple malignancies, including LIHC. Elevated DDX52 expression correlated with poor prognosis in LIHC. Genetic analysis revealed frequent copy number variations (CNV) and single-nucleotide variants (SNV) of DDX52, with significant positive correlations between DDX52 expression and both TMB and MSI across numerous cancer types. DDX52 expression was also significantly associated with immune cell infiltration, immune checkpoint molecules. In LIHC, DDX52 was predominantly expressed in B cells, proliferating T cells, malignant cells, and monocytes/macrophages. Functional enrichment analysis indicated DDX52 involvement in metabolic processes, histone modification, cell cycle regulation, and oncogenic signaling pathways. Pharmacologically, high DDX52 expression correlated with reduced drug sensitivity. Experimentally, DDX52 silencing significantly inhibited proliferation, colony formation, migration, and invasion in HepG2 and MHCC-97H cells. CONCLUSIONS:Our integrative analyses and experimental validation suggest that DDX52 is a potential prognostic biomarker and therapeutic target in LIHC. Its associations with immune features and multiple cancer-related pathways provide hypotheses for future mechanistic investigation.
OBJECTIVE:Hinokiflavone (HF) exhibits multiple pharmacological activities. This study aimed to evaluate the effect of HF on cytochrome P450 (CYP450s), which may provide a theoretical basis for its rational clinical application. METHODS:Human pooled liver microsomes (HLMs) were incubated with HF and CYP isoform-specific probe substrates, with the vehicle group (no HF and positive inhibitors) serving as a negative control group and positive inhibitors as a positive control group. The inhibitory effect of HF on CYP2C9, 2C19, and 3A4 was evaluated by using HF at concentrations of 0, 2, 5, 10, 20, 50, and 100 μM. The inhibitory model and parameters of HF on CYP2C9, 2C19, and 3A4 were assessed by the Lineweaver-Burk plots, secondary linear and nonlinear regression analysis. RESULTS:Selective inhibition of HF was observed on CYP2C9, 2C19, and 3A4 among the eight main CYP enzymes. HF showed concentration-dependent inhibition on CYP2C9, 2C19, and 3A4 with the IC50 values of 6.79 μM, 13.59 μM, 3.52 μM for 3A4-T (testosterone as the substrate), and 3.95 μM for 3A4-M (midazolam as the substrate), respectively. Through the inhibitory model evaluation, HF was revealed to be a competitive inhibitor of 2C9 (Kᵢ = 2.951 μM) and 2C19 (Kᵢ = 5.958 μM). The inhibitory model of HF on CYP3A4 was a non-competitive and time-dependent inhibition with the Ki of 1.226 μM, KI of 1.410 μM, and Kinact of 0.095 min-1. CONCLUSIONS:HF competitively inhibited the activities of CYP2C9 and CYP2C19, whereas it exhibited non-competitive and TDI against CYP3A4. Concomitant use of HF with medications primarily metabolized by CYP2C9, CYP2C19, or CYP3A4 may pose a risk of clinically relevant drug-drug interactions (DDI).
Background: R-loops are three-stranded nucleic acid structures formed by a DNA-RNA hybrid and a displaced single-stranded DNA. They regulate transcription, replication, and DNA repair, but their dysregulation causes genomic instability and inflammation, contributing to brain diseases. The nervous system exhibits selective vulnerability to R-loop stress due to ultra-long gene transcription, post-mitotic longevity, and high metabolic demands. Methods: This review synthesizes current literature from PubMed, Scopus, Web of Science, and Embase (2010–2026) on R-loop biology, with a focus on brain-specific mechanisms, regulatory factors (SETX, ZPR1, METTL3, TDP-43/FUS), and disease models. Results: In neurodegeneration, R-loop accumulation drives repeat expansion disorders (Fragile X, Huntington's disease) and loss-of-function SETX mutations (AOA2), whereas gain-of-function SETX (L389S) causes pathological R-loop depletion in ALS4, disrupting TGF-β signaling. TDP-43/FUS and SMN are integral to R-loop resolution, unifying ALS/FTD and SMA. In brain cancers, METTL3-mediated m6A modification of TERRA stabilizes telomeric R-loops in ALT-positive neuroblastoma, creating a therapeutic vulnerability to METTL3 inhibitors (STM2457, STC-15). Glioma stem cells rely on m6A-modified circPOLR2B to regulate R-loop formation and malignancy. Clinical-stage agents (EP102, TUG1ASO, ATX-559) and R-loop-derived prognostic signatures (RLPI) are emerging, but translation is hindered by a lack of non-invasive biomarkers and the dual physiological/pathological roles of R-loops. Conclusions: R-loops are central to brain disease pathogenesis, offering promising therapeutic targets. Future research should prioritize precision R-loop modulators, non-invasive biomarkers, and combinatorial strategies.
TRAF1 is an NF-κB inducible signaling adaptor protein that regulates immune responses. While it limits the NF-κB and MAPK pathways downstream of TLR in monocytes and macrophages, it promotes the survival of T and B lymphocytes downstream of 4-1BB and CD40, respectively. This multifaceted role requires genetic and pharmacological manipulation of TRAF1 expression and its interaction with its signaling partners. Key to such studies is the availability of reliable and specific antibodies that detect TRAF1 protein expression. Here, we systematically evaluate four different commercially available TRAF1 antibody clones (1F3, H3, E12, and 45D3) using wildtype (WT) and Traf1 knockout (KO) cells as definitive controls. We confirm robust induction of Traf1 mRNA in wild-type bone marrow derived macrophages (BMDMs) following stimulation with LPS, Pam3CSK4, or Resiquimod, and T cell activation by CD3 or CD3 + CD28 co-stimulation. Despite this, clones 1F3 and 45D3 produced non-specific signals in negative-control lysates, while clones E12 and H3 detected mouse TRAF1 only when overexpressed in HEK293FT cells and failed to detect endogenous TRAF1 in isolated BMDMs or T cells, even when lysates were intentionally overloaded or enriched by immunoprecipitation. Notably, clone H3 reproducibly detected TRAF1 in lysates of total splenocytes activated with anti-CD3 or anti-CD3 plus LPS, indicating that the assay itself is functional and that detection becomes possible in mixed cell populations where higher-expressing subsets are present. These findings reveal a critical sensitivity gap: currently available commercial antibodies are not adequate for measuring endogenous TRAF1 in isolated monocytes/macrophages or T cells, the very cell types most relevant to inflammation-driven and autoimmune diseases, and underscore the need for new, rigorously validated reagents to support cell-type specific studies of TRAF1 biology in mouse models.
Tumor-immune interactions in the cancer microenvironment have a major influence on the development of colorectal cancer (CRC), as well as immune evasion by the CRC tumor. Chemokine CXCL9 (which signals through the CXCR3 receptor) has been implicated in the immune recruitment of cells and progression of tumors; however, the role of CXCL9 in CRC is poorly understood. To explore the mechanistic role of CXCL9 in CRC cell proliferation, migration, invasion, survival, and immune modulation, we utilized CRC cell lines HCT116 and HT-29. Knocking down CXCL9 expression using siRNA decreased CRC cell proliferation, EdU incorporation, clonogenic survival, and anchorage-independent growth. The restoration of CXCL9 expression partially reversed these observations. In migration and invasion assays, CXCL9 increased motility and epithelial-mesenchymal transition (EMT) of CRC cells via the regulation of E-cadherin, N-cadherin, and vimentin expression. Silencing CXCL9 expression (due to the CXCL9 siRNA) resulted in increased apoptosis and G0/G1 cell cycle arrest, whereas restoring CXCL9 expression allowed CRC cells to transition to S-phase. Mechanistically, CXCL9 maintained redox homeostasis by preventing accumulation of reactive oxygen species (ROS), and activating pro-survival signaling pathways (AKT, ERK1/2, and NF-κB). Importantly, CXCL9 positively regulated immune checkpoint molecules (PD-L1 and IDO1) at both mRNA and protein levels. In co-culture experiments, CXCL9 promoted CD8+ T-cell chemotaxis through CXCR3 and enhanced T-cell-mediated cytotoxicity, effector molecule expression, and pro-inflammatory cytokine secretion under the tested in vitro conditions. Together, these in vitro findings suggest that CXCL9 contributes to CRC cell proliferation, migration, invasion, redox regulation, immune checkpoint-associated signaling, and CD8+ T cell responses in simplified co-culture models. The novelty of this study lies in the combined evaluation of tumor-cell phenotypes, PD-L1/IDO1-associated signaling, and CD8+ T cell responses within the same CXCL9-modulated CRC cell-line system. Further validation using patient-derived samples, in vivo models, and clinically annotated cohorts is required before broader conclusions regarding the CRC tumor microenvironment or therapeutic targeting can be made.
BACKGROUND:The low early diagnostic efficacy of non-small cell lung cancer (NSCLC) is a key contributor to its high mortality rate, and small nucleolar RNAs (snoRNAs) in serum exosomes can serve as a beneficial liquid biopsy approach for the early diagnosis of NSCLC. METHODS:Exosomes were isolated from collected serum; their morphology was imaged using transmission electron microscopy (TEM); particle size was measured using a particle size analyzer; and the expression of exosomal membrane proteins was identified using Western blot. Gene chips were used to screen for differentially expressed snoRNAs in exosomes, which were further validated by quantitative PCR (qPCR). The area under the receiver operating characteristic (ROC) curve (AUC) was used to estimate their diagnostic performance for NSCLC. Their biological functions in NSCLC were evaluated using an in vitro study. RESULTS:A series of exosome characterization experiments confirmed successful exosome extraction. Microarray and qPCR analyses revealed that serum exosomal snoRNAs (AC092799.1-201 and AC009408.1-201) were significantly upregulated in individuals with NSCLC. When combined with CEA and CYFRA21-1, these two exosomal snoRNAs achieved diagnostic efficacy of 0.948 and early diagnostic efficacy of 0.917. Cell experiments confirmed that AC092799.1-201 is related to rapid proliferation and high invasiveness of tumor cells. CONCLUSION:Exosomal snoRNAs AC092799.1-201 and AC009408.1-201, merged with CEA and CYFRA21-1, can serve as a novel liquid biopsy approach for the early diagnosis of NSCLC.
Celiac disease (CD) is a prototypical example of gluten-induced enteropathy characterized by a strong breakdown of mucosal tolerance. However, the spatial processes underlying intercellular communication across the epithelial-lamina propria barrier remain poorly known. Exosomes have evolved to be useful orchestrators of this autoinflammatory cascade. In contrast to their passive metabolic by-product role, gut-derived exosomes functionally mediate the non-canonical presentation of deamidated gliadin peptides and disease-associated HLA-DQ complexes, which directly prime pathogenic CD4+ T cells independently of conventional cellular synaptic interaction. At the same time, dysregulated exosomal microRNA (miRNA) signatures undermine tight junction integrity and promote pro-inflammatory signaling loops systemically, providing a probable mechanistic link to extra-intestinal symptoms. In this review, the dual role of exosomes as pathogenic propagation vectors and real-time liquid biopsies in CD will be critically discussed.
Acute myeloid leukemia (AML) is a hematologic malignancy that necessitates the identification of new therapeutic targets. Recently, the low-density lipoprotein receptor (LDLR) has been linked to an unfavorable prognosis in AML. LDLR plays a crucial role in various signaling pathways, including the MAPK signaling pathway. Our data show high LDLR expression in the AML cell lines THP-1 and NB4. Knockdown of LDLR using shRNA reduced AML cell proliferation, induced apoptosis, and led to S-phase cell accumulation, accompanied by modulation of the MAPK pathway. Additionally, myosin regulatory light chain-interacting protein (MYLIP) is expressed at low levels in AML and regulates LDLR. Overexpression of MYLIP downregulated LDLR expression, accompanied by an influence on the MAPK signaling pathway. Functionally, MYLIP overexpression decreased AML cell proliferation, increased apoptosis, and potentially delayed S-phase progression. These findings suggest that MYLIP and LDLR may regulate AML cell expansion, potentially through effects on the MAPK pathway. Furthermore, the p38-MAPK activator PCS reversed the inhibitory effect of LDLR knockdown and MYLIP overexpression on cell proliferation. Consequently, targeting LDLR could be a promising approach for AML treatment.
Multiple myeloma (MM) remains largely incurable despite major therapeutic advances, underscoring the need to define novel pathogenic mechanisms and druggable targets. Epitranscriptomic dysregulation, encompassing reversible chemical modifications on RNA, has emerged as a post-transcriptional regulatory layer that may contribute to MM biology. This focused review discusses the emerging roles of major RNA modifications and their regulators in MM pathogenesis, bone disease, drug resistance, and immune escape. We summarize representative experimental and translational studies on RNA-modifying enzymes, non-coding RNAs, and the bone marrow microenvironment, with emphasis on mechanisms directly validated in MM. Evidence derived from AML, solid tumors, or pan-cancer analyses is discussed as hypothesis-generating and requiring MM-specific validation. We summarize MM-supported evidence that m6A demethylases such as FTO and ALKBH5, as well as writers such as METTL3 and NSUN2, may regulate the stability and translation of disease-relevant transcripts. We also discuss emerging cross-cancer data on the m7G writer METTL1 as a hypothesis-generating framework that requires MM-specific validation. We delineate how RNA modification-dependent non-coding RNA networks and extracellular vesicle cargo remodel osteoclast and osteoblast function, linking the epitranscriptome to osteolytic bone disease. We further describe RNA modification-driven drug resistance circuits and immune escape pathways involving FTO, METTL3, H19, MALAT1, YTHDF1, and m5C-defined molecular subtypes. Finally, we summarize current epitranscriptomic therapeutic strategies, including small molecule inhibitors of writers, erasers, and readers, RNA-based therapeutics targeting pathogenic non-coding RNAs, and RNA modification-derived prognostic signatures for risk stratification. Collectively, this review discusses RNA-modification machinery as a potentially actionable regulatory layer in MM and outlines key challenges for clinical translation.
Formalin-fixed paraffin-embedded (FFPE) tissues are among the most commonly preserved specimen types in pathology. Clinical-grade next-generation sequencing (NGS) tests require extensive method development including nucleic acid extraction, analytic and clinical validation, and independent laboratory verification studies. The recently FDA-approved Oncomine Dx Express Test (ODxET) is an in vitro diagnostic (IVD) amplicon-based NGS test designed for clinical testing. This study evaluated clinical accuracy of the test in an external laboratory from the manufacturer and using an independent reference assay. We assessed 48 serially sectioned FFPE tissue from 13 diverse tumor types using ODxET on the Genexus Integrated System as compared to a reference hybrid capture NGS panel processed at an independent laboratory. Discrepant findings were confirmed with either a validated amplicon-based NGS panel, or with droplet digital polymerase chain reaction (ddPCR). Overall concordance was 98% at the variant level (n = 89/91) and 96% at the sample level (n = 45/47), with VAFs ranging from 0.37 to 87.51%. Transfer of ODxET into the clinical laboratory was executed following the current guidance from the CAP organization. Standardization of procedures, optimization of NGS assays, and harmonization of bioinformatics pipelines critically improved the inter-laboratory comparability of genomic profiling from FFPE tissue samples and allowed for rapid integration into the clinical laboratory.
Epigenetic silencing of tumor suppressor genes (TSGs) is a key driver of colorectal cancer (CRC) progression, with phosphatase and tensin homolog deleted on chromosome 10 (PTEN) frequently inactivated through promoter hypermethylation. This study explored the association between DNA methyltransferase 1 (DNMT1) suppression, PTEN reactivation, and miR-29a-3p modulation following 5-aza-2'-deoxycytidine (5-aza-dC) treatment in CRC cell models. Bioinformatic analyses revealed DNMT1 overexpression, increased PTEN promoter methylation, and a potential regulatory network associated with miR-29a-3p. In vitro, HCT116 and SW620 cells treated with 5-aza-dC exhibited dose-dependent reductions in cell viability, with IC50 values of 12.38 ± 0.98 μM and 10.04 ± 0.98 μM, respectively. Treatment with 5-aza-dC induced significant PTEN re-expression and DNMT1 downregulation at both the mRNA and protein levels. Methylation-specific PCR (MSP-PCR) suggested reduced PTEN promoter methylation following treatment. Additionally, miR-29a-3p expression was significantly upregulated. These findings suggest that 5-aza-dC restores PTEN expression in association with DNMT1 suppression and PTEN promoter demethylation, while miR-29a-3p upregulation may reflect an associated epigenetic response requiring further mechanistic validation.
Previous research has indicated that Zinc finger MIZ domain-containing protein 1 (ZMIZ1) was elevated in patients with acute myeloid leukemia (AML); however, the pathological mechanisms underlying ZMIZ1 up-regulation in AML remain unclear. The expression levels of the target proteins, PAX6 ubiquitination modifications, and protein stability were evaluated by Western blot. The effects of regulating ZMIZ1 expression on the growth of AML cells were detected through in vitro and in vivo experiments. The regulatory effects of ZMIZ1 on SMAD3 and PAX6, as well as rescue experiments, were conducted in HL60 cells. In this study, we found that ZMIZ1 and SMAD3 were upregulated in patients with AML, while PAX6 was downregulated. Overexpression of ZMIZ1 significantly promoted HL60 cell growth, while inhibiting cell apoptosis. Consistent with these observations, up-regulation of ZMIZ1 markedly promoted tumor cell growth in nude mice, manifested as increased tumor volume and weight, as well as enhanced tumor cell proliferation. However, ZMIZ1 down-regulation had opposite effects both in vitro and in vivo. Mechanistically, PAX6 is a downstream regulatory factor of ZMIZ1, and its ubiquitination at the K53 position is regulated by ZMIZ1. SMAD3 acts as a bridge to mediate the ubiquitination modification of ZMIZ1 on PAX6. More importantly, PAX6 up-regulation suppressed cell growth induced by ZMIZ1 or SMAD3 overexpression. In conclusion, our findings suggest that targeting ZMIZ1 or overexpressing PAX6 serve as a therapeutic strategy for AML.
Background Wound healing is a complex biological process involving vascular remodeling and tissue repair. However, the mechanism by which granulocyte-macrophage colony-stimulating factor (GM-CSF) facilitates vascular maturation during wound healing remains unclear. This study aims to investigate the biological mechanism by which GM-CSF promotes vascular maturation and enhances wound repair. Methods Human dermal microvascular endothelial cells (ECs) and human brain vascular pericytes (PCs) were cultured separately or in co-culture and treated with various concentrations of GM-CSF. Cell proliferation and migration were assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylformazan and Transwell assays. Tube formation assays were performed to evaluate angiogenic potential. The expression of vascular endothelial growth factor (VEGF) and angiopoietin-1 (Ang-1) was measured by enzyme-linked immunosorbent assay (ELISA) and Western blot. Endothelial permeability was analyzed using fluorescein isothiocyanate (FITC)-dextran staining. The expression of platelet endothelial cell adhesion molecule 1 (PECAM-1) was assessed by immunofluorescence. Results GM-CSF substantially raised EC and PC proliferation and migration in a dose-dependent manner, with more pronounced effects observed in co-culture conditions. GM-CSF grew VEGF and Ang-1 in PCs and enhanced tube formation and barrier integrity in both mono- and co-cultured ECs. Furthermore, GM-CSF-stimulated PC-conditioned medium induced the upregulation of PECAM-1 in ECs. Conclusion GM-CSF promotes PECAM1 expression, angiogenesis, and vascular maturation in ECs by upregulating VEGF and Ang-1 in PCs. These effects enhance the interaction between endothelial cells and pericytes, thereby contributing to vascular stabilization and improved wound healing.
Exosomes, a specialized class of extracellular vesicles, exhibit significant therapeutic potentials for neurological disorders, in particular for vascular dementia (VaD). VaD is the second most common form of dementia, characterized by cognitive and behavioral impairments. VaD is often linked to hippocampal damage resulting from its vulnerable vascular structure, which disrupts memory formation and retrieval. Secreted by various cell types within the central nervous system, exosomes mediate intercellular communication by transporting bioactive molecules. Growing evidence indicates that exosomes enhance synaptic plasticity, modulate neuroinflammation, inhibit apoptosis, and promote angiogenesis, supporting their therapeutic potentials in VaD. Given the urgent need for effective treatments and the unique ability of exosomes to cross the blood-brain barrier (BBB) and deliver multi-targeted therapies, research in this field is critically important. It offers a viable pathway toward the development of disease-modifying interventions for a condition that is currently managed primarily through symptomatic treatment. This review summarizes current knowledge on the functions of exosomes in the central nervous system, explores recent advances in exosome-based strategies for VaD, and discusses ongoing challenges and future directions for their clinical translation.
Background: Non-small cell lung cancer (NSCLC) is the most common subtype of lung cancer with a steady increase in incidence. Purpose: This study aimed to confirm the role of LINC01583 on NSCLC progression and Osimertinib (Osi) resistance. Methods: The interactions of LINC01583, miR-4640-5p, and CHD8 were confirmed by RNA pulldown and Luciferase assay. The ROC curve was utilized to assess the discriminatory ability of LINC01583 for patients with Osi-resistant. NSCLC cell lines H1975 and PC9 were selected, and Osi-resistant cell lines were established by drug administration using the concentration gradient escalation method. The effect of LINC01583 on lung cancer drug-resistant cell lines was evaluated using CCK8, Transwell assay, and rescue experiments. Results: In NSCLC cell lines and patients, the expression of LINC01583 and CHD8 was significantly increased, while miR-4640-5p expression was significantly decreased. Mechanistically, knockout of LINC01583 suppressed the migration ability of Osi-resistant cells and increased their sensitivity to Osi. Upregulation of LINC01583 can significantly promote the migration ability of lung cancer cell parent lines and reduce their sensitivity to Osi. Rescue experiments showed that co-transfection of miR-4640-5p inhibitor partially reversed the inhibitory effects of LINC01583 knockdown. LINC01583 acted as a molecular sponge for miR-4640-5p and regulated CHD8 expression by competitive binding. Conclusion: In this study, we revealed LINC01583 as a key oncogenic driver that promotes both NSCLC progression and Osi resistance by functioning as a competing endogenous RNA (ceRNA) for miR-4640-5p, thereby upregulating its target gene CHD8.
Myocardial Hypoxia-Reperfusion (H/R) injury is a major challenge in coronary artery disease treatment. This article explores the potential mechanism of salt inducible kinase 1 (SIK1) in H/R injury. This study used H9C2 cardiomyocytes as the model. By constructing SIK1 overexpression and myocyte enhancer factor 2 (MEF2) silencing vectors, and combining qRT-PCR, CCK8, 5-Ethynyl-2'-deoxyuridine, flow cytometry, Western blot, JASPAR prediction, luciferase assay and chromatin immunoprecipitation experiments, it explored the effects and mechanisms of SIK1 on H/R cardiomyocytes from multiple dimensions. H/R impaired H9C2 cells (reduced viability, increased apoptosis/ROS/Caspase-9/12/Cytochrome C, downregulated MEF2/Sirtuin 1 (SIRT1)), which SIK1 overexpression reversed. MEF2 silencing blocked SIK1's protection; JASPAR, luciferase assay and Chromatin Immunoprecipitation confirmed MEF2 directly bound to SIRT1 promoter and activated its transcription. SIK1 alleviates H/R-induced myocardial cell damage in association with the upregulation of MEF2 and subsequent MEF2-mediated transcriptional activation of SIRT1.
