Biology Direct[JOURNAL]
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200 papers
Contactin-2 mitigates heart failure and cardiac remodeling via regulation of NUPR1 and ferroptosis.
Fei YD, Wei ZX, Chen TZ … +21 more , Zhang LH, Zhang YC, Hou JW, Wang Q, Cai XX, Chen M, Yang YL, Yao PC, Fei ZT, Bai Y, Zhao TT, Yan J, Li W, Yang M, Zhang R, Liu XD, Zhao MZ, Xu XL, Sun J, Wang YP, Li YG
Biol Direct · 2026 Jul · PMID 42401981 · Full text
BACKGROUND: The roles of Contactin-2 (CNTN2) and ferroptosis in heart failure and cardiac remodeling remain incompletely understood. RESULTS: CNTN2 was significantly upregulated in hypertrophic cardiomyopathy patients an...
BACKGROUND: The roles of Contactin-2 (CNTN2) and ferroptosis in heart failure and cardiac remodeling remain incompletely understood. RESULTS: CNTN2 was significantly upregulated in hypertrophic cardiomyopathy patients and heart failure mice. In cardiomyocyte specific CNTN2 conditional knockout (CNTN2 cKO) mice, transverse aortic constriction (TAC) induced markedly exacerbated heart failure, cardiac remodeling and ferroptosis compared to control mice. Ferroptosis inhibition substantially attenuated heart failure in CNTN2 cKO mice subjected to TAC, indicating that enhanced ferroptosis contributes to the detrimental effects of CNTN2 deficiency. RNA sequencing identified NUPR1, a ferroptosis repressor, as a downstream molecule of CNTN2. Mechanistically, CNTN2 activated the Lyn/eIF2α/ATF4 pathway to regulate NUPR1. CNTN2 overexpression attenuated Angiotensin II-induced cardiomyocyte ferroptosis and pathological remodeling, whereas these protective effects were abolished by Lyn or NUPR1 inhibitors. We further revealed CNTN2 and Lyn interacted with each other, and that CNTN2 interacted with Lyn through its 1-328aa domain. In vivo NUPR1 overexpression via AAV9 significantly mitigated TAC-induced heart failure and cardiac remodeling in CNTN2 cKO mice. CONCLUSIONS: Our study demonstrates that CNTN2 protects against pressure overload induced heart failure and cardiac remodeling by regulating ferroptosis through the Lyn/eIF2α/ATF4/NUPR1 pathway, suggesting CNTN2 as a potential therapeutic target.
LINC00324 induces nucleus pulposus cell apoptosis in intervertebral disc degeneration via miR-143-3p targeted TRAIL/DR4/DR5 axis.
Zhang J, Zhao M, Shan Z … +2 more , Chang Y, He Y
Biol Direct · 2026 Jul · PMID 42401954 · Full text
BACKGROUND: Intervertebral disc degeneration (IDD) serves as a predominant pathogenic driver of spinal degenerative diseases and low back pain. Long non-coding RNAs (lncRNAs) have emerged as important regulators involved...
BACKGROUND: Intervertebral disc degeneration (IDD) serves as a predominant pathogenic driver of spinal degenerative diseases and low back pain. Long non-coding RNAs (lncRNAs) have emerged as important regulators involved in the pathogenesis of IDD. Despite this, the specific contributions of LINC00324, a particular lncRNA, to the advancement of IDD are not yet well understood. The study seeks to elucidate the functional role and explore the potential molecular mechanisms of LINC00324 in IDD. METHODS: RT-qPCR was employed to analyze LINC00324, miR-143-3p, and TRAIL levels in intervertebral disc (IVD) tissues from IDD patients and control subjects, followed by correlation analysis. Bioinformatics tools, dual-luciferase reporter assays, and RNA pull-down experiments were conducted to validate the regulatory interactions within LINC00324/miR-143-3p/TRAIL in nucleus pulposus cells (NPCs). In vitro, NPCs were stimulated with lipopolysaccharide (LPS) to construct an IDD cell model. RT-qPCR, western blot, CCK-8 assay, and flow cytometry were utilized to evaluate the effects on the levels of LINC00324/miR-143-3p/TRAIL/DR4/DR5, as well as the proliferation and apoptosis of NPCs. Additionally, extracellular matrix (ECM)-related proteins and inflammatory factors were detected by western blot and ELISA, respectively. In vivo, a rat IDD model was established through fibroannular puncture. Radiological analysis (X-ray), HE staining, TUNEL staining, and western blot were used to assess disc structural changes, NPC apoptosis, and the levels of miR-143-3p/TRAIL/DR4/DR5 and apoptosis-related proteins. RESULTS: Clinical evidence showed that LINC00324 and TRAIL levels were increased, and miR-143-3p was decreased in IDD samples. miR-143-3p had the negative correlation coefficient with TRAIL and LINC00324. Furthermore, LINC00324 acted as a sponge for miR-143-3p, and miR-143-3p specifically attenuated the downstream of TRAIL level. In vitro, the LPS-induced IDD model showed decreased NPC viability, increased apoptosis, disrupted ECM metabolism (downregulated Collagen II and Aggrecan, upregulated MMP13), elevated inflammatory factors, and activated TRAIL/DR4/DR5 axis. Silencing LINC00324 or overexpressing miR-143-3p alleviated these pathological changes. Furthermore, miR-143-3p partially counteracted the promoting influences of LINC00324 overexpression on NPC apoptosis, ECM degradation, inflammatory response, and TRAIL/DR4/DR5 axis activation in the IDD model. In vivo, LINC00324 overexpression exacerbated IDD in rats (decreased disc height index, disrupted NPC tissue structure, enhanced NPC apoptosis, aggravated ECM degradation, inhibited miR-143-3p expression, and activated TRAIL/DR4/DR5 axis), while LINC00324 knockdown attenuated these degenerative changes. CONCLUSIONS: In conclusion, LINC00324 promotes IDD progression by inducing NPC apoptosis, disrupting ECM metabolism, and enhancing inflammatory response via sponging miR-143-3p to activate the TRAIL/DR4/DR5 axis. CLINICAL TRIAL NUMBER: Not applicable.
Epicatechin suppresses cancer stem-like cell phenotype and radioresistance in non-small cell lung cancer via targeting CXCL8.
Li T, Han H, Liu M … +11 more , He Y, Zhou S, Xiao Y, Huang N, Peng N, Li C, Xu H, Xia J, Lv Z, Wu Y, Huang M
Biol Direct · 2026 Jul · PMID 42401948 · Full text
BACKGROUND: Radiotherapy remains a cornerstone for non‑small cell lung cancer (NSCLC). However, its efficacy is frequently limited by radioresistance, a phenomenon closely associated with cancer stem-like cells (CSCs). A...
BACKGROUND: Radiotherapy remains a cornerstone for non‑small cell lung cancer (NSCLC). However, its efficacy is frequently limited by radioresistance, a phenomenon closely associated with cancer stem-like cells (CSCs). Although our previous work demonstrated that epicatechin (EC) sensitizes NSCLC cells to ionizing radiation (IR), whether this effect involves modulation of CSCs and the underlying mechanisms remain to be elucidated. METHODS: A radioresistant NSCLC cell line (A549RR) was established via fractionated irradiation and employed in both in vitro assays and murine xenograft models. To assess its radiosensitizing potential, EC was administered prior to IR exposure. Cell viability and colony formation capacity were measured to evaluate radiosensitivity. Putative targets of EC were predicted using network pharmacology and validated through molecular docking. The direct interaction between EC and CXCL8 was examined by cellular thermal shift assay (CETSA). The functional relevance of CXCL8 in EC-mediated effects was assessed through both loss-of-function (shRNA-mediated knockdown) and gain-of-function (CXCL8 overexpression) approaches. CSC properties were evaluated by tumor sphere formation assays and immunoblotting for stemness markers. RESULTS: EC significantly enhanced NSCLC radiosensitivity both in vitro and in vivo, concomitant with marked suppression of CSC stemness. CXCL8 was identified as a direct functional target of EC: its expression was substantially upregulated in radioresistant cells but downregulated upon EC treatment. CETSA confirmed a direct interaction between EC and CXCL8 in cells. Functionally, CXCL8 knockdown suppressed cell viability, colony formation capacity, and CSC properties in A549RR cells. Conversely, overexpression of CXCL8 abrogated EC-induced radiosensitization and restored CSC phenotypes in both cellular and xenograft models. CONCLUSION: EC overcomes NSCLC radioresistance by directly targeting CXCL8, thereby disrupting CSC-like traits.
MLKL confers ferroptosis resistance in colorectal cancer cells via impaired ESCRT-III-mediated membrane repair.
Sonkaew S, Duangthim N, Butkinaree C … +4 more , Yingchutrakul Y, Choksi S, Liu ZG, Jitkaew S
Biol Direct · 2026 Jul · PMID 42401932 · Full text
BACKGROUND: Ferroptosis resistance is a major challenge in cancer therapy, contributing to poor prognosis and unsatisfactory treatment outcomes. Thus, identifying key regulators of ferroptosis is essential for developing...
BACKGROUND: Ferroptosis resistance is a major challenge in cancer therapy, contributing to poor prognosis and unsatisfactory treatment outcomes. Thus, identifying key regulators of ferroptosis is essential for developing strategies to overcome this limitation. Mixed lineage kinase domain-like protein (MLKL) is well established as the executioner of necroptosis, but emerging evidence suggests broader functions beyond necroptosis. A previous study demonstrated that MLKL regulates ferroptosis in an acute kidney injury model. However, its role in ferroptosis regulation in cancer remains largely unexplored. RESULTS: Bioinformatics analyses revealed that MLKL expression positively correlates with glutathione metabolism-related genes in colorectal cancer (CRC) patients and with reduced sensitivity to GPX4 inhibitors across pan-cancer cell lines. Functional studies demonstrated that MLKL depletion sensitizes not only CRC cells but also multiple cancer cell types to ferroptosis induced by GPX4 inhibitors. Mechanistically, MLKL-deficient CRC cells exhibited reduced intracellular glutathione levels and dysregulation of the NRF2/KEAP1/SLC7A11 antioxidant axis. However, KEAP1 knockdown in MLKL-depleted cells partially reduced ferroptosis sensitivity, indicating that additional mechanisms are involved. Proteomic analysis identified CHMP5, a component of the ESCRT-III membrane repair machinery, as a key downstream effector associated with MLKL. MLKL depletion suppressed CHMP5 expression and impaired its translocation to the plasma membrane during ferroptosis induction. Consistent with defective membrane repair, MLKL-knockdown CRC cells exhibited increased HMGB1 release following GPX4 inhibitor treatment, indicating enhanced membrane damage. Notably, CHMP5 overexpression reduced ferroptosis in MLKL-knockdown cells, highlighting that MLKL regulates ferroptosis susceptibility in part through CHMP5-dependent ESCRT-III signaling. CONCLUSIONS: This study reveals a previously unrecognized role for MLKL in ferroptosis regulation through coordinating NRF2/KEAP1/SLC7A11-mediated glutathione metabolism and ESCRT-III-dependent membrane repair. These findings identify MLKL as a potential regulator of ferroptosis resistance and warrant further studies to evaluate the therapeutic relevance of MLKL-targeting strategies in colorectal cancer.
Single-cell RNA sequencing reveals lipid metabolism disorders in the retina in spontaneous high myopia.
Bao B, Liu J, Xie Y … +8 more , Xi R, Sun Y, Yan J, Xin J, Yang Z, Yin X, Guo D, Bi H
Biol Direct · 2026 Jul · PMID 42393743 · Full text
BACKGROUND: Myopia is one of the most common eye diseases affecting children and adolescents, with its etiology often attributed to a combination of genetic and environmental factors. This study utilized single-cell RNA...
BACKGROUND: Myopia is one of the most common eye diseases affecting children and adolescents, with its etiology often attributed to a combination of genetic and environmental factors. This study utilized single-cell RNA sequencing (scRNA-seq) technology to investigate gene expression differences between guinea pigs with spontaneous high myopia (SHM) and those with normal vision. RESULTS: Lens-induced myopia (LIM) and SHM guinea pigs exhibit significant retinal structural abnormalities and functional impairments, characterized by reduced retinal thickness and abnormal electroretinogram (ERG) responses, indicating progressive retinal dysfunction during myopia development. scRNA-seq and molecular experiments revealed that the ceramide synthase 5 (CERS5) / ceramide synthase 6 (CERS6) / alkaline ceramidase 3 (ACER3) / phospholipid phosphatase 1 (PLPP1) / prosaposin (PSAP) / UDP-glucose ceramide glucosyltransferase (UGCG) signaling axis is significantly activated in the retinas of myopic guinea pigs, suggesting dysregulation of sphingolipid metabolism. These changes were associated with an increased expression of SPHK1 and a shift in receptor expression from S1PR1 to S1PR2, indicating a transition in lipid signaling from a physiological state to a stress-related pathological state. Additionally, the activation of the P62/NRF2/KEAP1 pathway confirmed enhanced oxidative stress in the myopic retina. This study demonstrates that the CERS5/CERS6/ACER3/PLPP1/PSAP/UGCG signaling axis mediates disruption of sphingolipid metabolism and oxidative stress in the myopic guinea pig retina. CONCLUSIONS: The present study demonstrates that the CERS5/CERS6/ACER3/PLPP1/PSAP/UGCG signaling axis mediates sphingolipid metabolic dysregulation and oxidative stress in the retinas of myopic guinea pigs. Mechanistically, the abnormal activation of the SPHK1-S1P signaling pathway, the shift in receptors from S1PR1 to S1PR2, and the subsequent activation of the P62/KEAP1/NRF2 pathway collectively led to changes in retinal morphology (such as reduced thickness) and functional impairment. These findings establish a mechanistic link between lipid metabolic imbalance and retinal pathology in myopia offering potential therapeutic targets for preventing retinal damage associated with myopia.
Running exercise alleviates chronic heart failure by promoting cardiomyocyte autophagic flux through the NEAT1-QKI affecting Beclin1/LC3B mRNA stability.
Zhang Y, Song R, Zhuang L … +6 more , Zhao Y, Chen Z, Wang Y, Mi S, Liu J, Tai W
Biol Direct · 2026 Jun · PMID 42374562 · Full text
As the end-stage manifestation of cardiovascular diseases (CVDs), chronic heart failure (CHF) is associated with high morbidity and mortality. Our previous study showed that Running exercise could improve CHF by inducing...
As the end-stage manifestation of cardiovascular diseases (CVDs), chronic heart failure (CHF) is associated with high morbidity and mortality. Our previous study showed that Running exercise could improve CHF by inducing autophagy, but the underlying mechanisms and applicability remain unclear. This study investigated the roles of NEAT1 and RNA-binding protein Quaking (QKI) in the cardioprotective effects of running exercise initiated during the early stage after abdominal aortic coarctation (AAC) surgery. NEAT1 and QKI were overexpressed in vivo or NEAT1 and QKI were knocked down in vitro. The effects of running exercise on the myocardial damage of CHF rats or Ang II-induced cardiomyocyte apoptosis, autophagic flux, autophagy-related protein expression, and the expression of the PI3K/ AKT pathway by regulating NEAT1 and QKI were assessed using echocardiography, tissue staining, immunofluorescence, western blotting, and confocal microscopy. Furthermore, RNA immunoprecipitation (RIP), RBPmap prediction, actinomycin D (ActD) chase experiments and Dual-luciferase reporter assays were performed to explore the post-transcriptional regulation of autophagy-related genes by QKI. Running exercise reduced myocardial tissue damage in CHF rats and up-regulated the expression of autophagy-related proteins. NEAT1 was up-regulated in the myocardial tissue of CHF rats, and Ang II-damaged cardiomyocytes. Whereas QKI expression showed the opposite trend. Additionally, running exercise inhibited NEAT1 while restoring QKI expression, particularly during weeks 2-3 of exercise intervention. Mechanistically, NEAT1 directly interacted with QKI and negatively regulated its expression. Simultaneously, QKI bound to the 3'untranslated regions (3'-UTRs) of Beclin1 and LC3B mRNA and enhances the stability of Beclin1 and LC3B transcripts, thereby promoting autophagy. Functionally, NEAT1 inhibition or QKI upregulation suppressed the PI3K/AKT/mTOR signaling pathway, and attenuated cardiomyocyte apoptosis under pathological conditions. Collectively, the current findings revealed the regulatory mechanism by which running exercise alleviates CHF, and clarified that running promotes cardiomyocyte autophagy and alleviates CHF through the NEAT1-QKI-Beclin1/LC3B regulatory axis, highlighting a novel mechanism underlying exercise-mediated cardioprotection. Given that exercise intervention was initiated during the early stage following AAC surgery, the observed benefits may include early remodeling-modifying and disease-progression-modifying effects, rather than solely therapeutic effects on established decompensated CHF.
The PTHR1/PKA/CREB1 axis promotes osteosarcoma progression by activating the PVT1/miR-590-3p/AXIN2 ceRNA network to induce epithelial-mesenchymal transition.
Zhang J, Li W, Liu F … +2 more , Zhang H, Wang W
Biol Direct · 2026 Jun · PMID 42374479 · Full text
BACKGROUND: Osteosarcoma (OS) remains a clinically challenging primary bone tumor because of its strong metastatic potential and unsatisfactory outcomes in advanced cases. Parathyroid hormone receptor 1 (PTHR1), a recept...
BACKGROUND: Osteosarcoma (OS) remains a clinically challenging primary bone tumor because of its strong metastatic potential and unsatisfactory outcomes in advanced cases. Parathyroid hormone receptor 1 (PTHR1), a receptor closely associated with bone-related signaling, has not been fully characterized in OS. This study explored whether PTHR1 contributes to OS progression and clarified the downstream regulatory mechanism involved. METHODS: Public GEO datasets, clinical OS specimens, and cultured OS cells were used to assess PTHR1 expression. Stable PTHR1 knockdown and overexpression OS cell models were established using lentiviral vectors. Cell growth, apoptosis, motility, invasiveness, EMT, and tumor growth were evaluated by CCK-8, flow cytometry, Transwell, Western blot, and xenograft assays. The underlying mechanism was explored using dual-luciferase reporter assays, ChIP-qPCR, pathway intervention, and rescue experiments. RESULTS: PTHR1 was significantly upregulated in OS tissues and cells. PTHR1 overexpression promoted OS cell proliferation, migration, invasion, EMT, and xenograft tumor growth while suppressing apoptosis, whereas PTHR1 knockdown exerted the opposite effects. Mechanistically, PTHR1 activated the cAMP/PKA/CREB1 pathway, leading to transcriptional upregulation of lncRNA PVT1. PVT1 functioned as a competing endogenous RNA by sponging miR-590-3p, thereby relieving miR-590-3p-mediated repression of AXIN2. AXIN2 further promoted EMT and malignant progression, and AXIN2 overexpression partially reversed the inhibitory effects of PTHR1 knockdown. CONCLUSION: PTHR1 promotes OS progression by activating the CREB1/PVT1/miR-590-3p/AXIN2 regulatory axis and enhancing EMT. This five-component signaling cascade may provide potential biomarkers and therapeutic targets for OS.
Identification and prognostic analysis of genes related to CTNNB1 mutations in hepatocellular carcinoma.
Zhu N, Zhang G, Shen X … +4 more , Chen Z, Li Y, Chen D, Jia H
Biol Direct · 2026 Jun · PMID 42363289 · Full text
BACKGROUND: Mutations in CTNNB1 are recognized oncogenic drivers of hepatocellular carcinoma (HCC); however, the downstream effector molecules and their prognostic significance remain incompletely defined. In this study,...
BACKGROUND: Mutations in CTNNB1 are recognized oncogenic drivers of hepatocellular carcinoma (HCC); however, the downstream effector molecules and their prognostic significance remain incompletely defined. In this study, we developed a mutant CTNNB1 gene signature (MCGS) to predict CTNNB1 mutation status and to evaluate the prognostic relevance of the constituent genes. METHODS: Hydrodynamic tail vein injection mouse models of HCC were generated by co-expressing Sleeping Beauty transposon/transposase and mutant CTNNB1 with either AKT-c-Myc or AKT-NRAS, followed by RNA sequencing of tumor tissues. Cross-species differential expression analyses were performed by integrating data from The Cancer Genome Atlas (TCGA) and the International Cancer Genome Consortium (ICGC). Machine-learning models were trained to identify MCGS. The prognostic value of signature genes was validated using public transcriptomic datasets and immunohistochemistry on an institutional tissue microarray (TMA) cohort. RESULTS: Mutant CTNNB1 markedly accelerated hepatocarcinogenesis in mice, leading to earlier tumor development and reduced survival. Transcriptomic profiling revealed enrichment of the Hippo and MAPK signaling pathways in CTNNB1-mutant HCCs. We established a 15-gene MCGS using a Categorical Boosting algorithm, which accurately predicted CTNNB1 mutation status (AUC = 0.957 in TCGA; 0.893 in ICGC). Among these genes, metallothionein-3 (MT3) emerged as a prognostic marker, with high expression independently associated with poorer overall survival in public datasets. Consistently, elevated MT3 protein expression on the TMA was also independently associated with worse overall and recurrence-free survival in our institutional cohort. CONCLUSION: We identified a robust mutant CTNNB1 gene signature capable of accurately predicting CTNNB1 mutation status, and uncovered MT3 as a novel, clinically relevant prognostic biomarker in HCC.
TrxR1 inhibition sensitizes hepatocellular carcinoma to Motesanib via an autophagy-ROS-JNK/ER stress axis.
Du S, Zheng P, Li W … +2 more , Chen G, Chen H
Biol Direct · 2026 Jun · PMID 42363247 · Full text
Hepatocellular carcinoma (HCC) remains a highly aggressive malignancy with a dismal prognosis, largely due to the limited effectiveness of current therapeutic interventions. Although Motesanib (MOT) is a clinically studi...
Hepatocellular carcinoma (HCC) remains a highly aggressive malignancy with a dismal prognosis, largely due to the limited effectiveness of current therapeutic interventions. Although Motesanib (MOT) is a clinically studied VEGFR-centered inhibitor, VEGFR-targeted strategies remain limited by innate or acquired resistance. In this study, we explored a novel pharmacological strategy combining MOT with Auranofin (AF) to enhance therapeutic outcomes in HCC. Our data demonstrate that this dual-targeting approach yields robust synergistic anticancer activity across diverse experimental models. Mechanistically, we discovered that TrxR1 inhibition is critical for this sensitization, which is closely associated with autophagy-related reactive oxygen species (ROS) accumulation. Specifically, genetic depletion of Atg5 effectively suppressed ROS accumulation and attenuated cytotoxicity, suggesting that autophagy contributes to the synergistic oxidative damage. This orchestrated stress response subsequently led to sustained endoplasmic reticulum (ER) stress and JNK pathway activation, culminating in DNA damage, impaired proliferation, and reduced cell viability. Taken together, our results identify the combination of MOT and AF as a promising and mechanistically grounded treatment regimen for advanced HCC (Graphical Abstract).
Integrated microbiome-metabolome analysis implicates Acinetobacter guillouiae in arachidonic acid metabolic remodeling and endometrial cancer cell proliferation.
Ma W, Pi J, Wan H … +4 more , Wang H, Han W, Hu M, Liu J
Biol Direct · 2026 Jun · PMID 42351301 · Full text
BACKGROUND: Lesion-associated microbiota have emerged as potential regulators of tumor biology. However, the ecological organization and metabolic relevance of the local endometrial microbiome in endometrial cancer (EC)...
BACKGROUND: Lesion-associated microbiota have emerged as potential regulators of tumor biology. However, the ecological organization and metabolic relevance of the local endometrial microbiome in endometrial cancer (EC) remain incompletely defined. This study aimed to characterize microbiome-associated metabolic alterations in EC tissues. METHODS: We performed integrated 16S rRNA sequencing and untargeted metabolomic analyses using paired EC and adjacent non-tumor endometrial tissues. Microbial diversity, taxonomic composition, co-occurrence network architecture, and metabolomic pathway alterations were evaluated. Conditioned medium derived from Acinetobacter guillouiae was used to explore its association with arachidonic acid (AA) metabolism and proliferative phenotypes in EC cells, followed by pharmacological pathway interrogation and xenograft validation. RESULTS: Overall microbial diversity was not significantly different between EC and adjacent tissues, whereas EC tissues exhibited a more connected and centralized microbial co-occurrence network. Taxonomic analyses identified enrichment of A. guillouiae in EC tissues based on 16S rRNA taxonomic assignment. Untargeted metabolomics revealed a group-level metabolic shift in EC tissues, with AA metabolism among the most prominently enriched pathways. In EC cells, A. guillouiae-derived conditioned medium increased intracellular AA levels, enhanced cPLA2 phosphorylation, induced AA-metabolizing enzymes, and activated the TLR4/NF-κB/ALOX5 signaling axis. Pharmacological inhibition of cPLA2, ALOX5, TLR4, or NF-κB partially attenuated the metabolic or proliferative effects associated with A. guillouiae conditioned medium. In a xenograft model, intratumoral exposure to A. guillouiae was associated with accelerated tumor growth. CONCLUSIONS: These findings suggest that A. guillouiae-associated microbial alterations may contribute to AA metabolic remodeling and proliferative phenotypes in EC. This study provides a microbiome-metabolome framework for understanding lipid metabolic heterogeneity in EC and supports further validation of bacteria-associated AA metabolism as a potential therapeutic target.
Comprehensive multi-omics analysis reveals a fatty acid metabolism gene signature for prognostic assessment and immunotherapy in nasopharyngeal carcinoma, and identifies ABCC1 as a potential novel therapeutic target.
Xu Y, Zhu L, Zhang Q … +9 more , Chen X, Lv H, Ma S, Wang R, Chang Y, Sun Y, Chen K, Li L, Zhu X
Biol Direct · 2026 Jun · PMID 42351260 · Full text
BACKGROUND: Nasopharyngeal carcinoma (NPC) is a subtype of head and neck squamous cell carcinoma characterized by high recurrence and metastasis rates and poor prognosis. Although immune checkpoint inhibitors have emerge...
BACKGROUND: Nasopharyngeal carcinoma (NPC) is a subtype of head and neck squamous cell carcinoma characterized by high recurrence and metastasis rates and poor prognosis. Although immune checkpoint inhibitors have emerged as a promising treatment strategy for recurrent/metastatic nasopharyngeal carcinoma (R/M NPC), only a few patients have benefitted significantly from them. Lipid metabolism reprogramming plays a crucial role in NPC progression and its interaction with the immune microenvironment. This study aims to establish a prognostic model for NPC based on lipid metabolism-related factors, further explore its association with tumor immunity, and investigate the potential for immunotherapy. METHODS: Collect GEO datasets(GSE53819 and GSE102349) for differential expression analysis and least absolute shrinkage and selection operator (LASSO) regression to identify prognostic genes and construct a fatty-acid-metabolism-related prognostic model. Survival analyses and time-dependent receiver operating characteristic (ROC) curves were applied to evaluate the predictive performance of the constructed prognostic markers. Furthermore, associations between the derived risk scores and immunological characteristics were systematically evaluated. Following the identification of ABCC1 as a key gene, its expression was validated through RT-qPCR, immunoblotting, and immunohistochemistry (IHC). Its functional role was further investigated using in vitro functional assays, multiplex immunohistochemistry (mIHC), co-culture experiments with CD8⁺ T cells, and in vivo xenograft tumor models in nude mice. RESULTS: Four genes (ABCC1, CD1D, CYP4B1, and DPEP2) were identified to constr uct a prognostic model associated with fatty acid metabolism. This model revealed significant distinctions in immune infiltration patterns between high-risk and low-risk groups. Specifically, the high-risk group displayed immunosuppressive characteristics, marked by reduced infiltration of CD8⁺T cells. Functional studies demonstrated that ABCC1 promoted NPC cell proliferation, migration, invasion, ROS accumulation, and lipid metabolic reprogramming. Mechanistically, ABCC1 was epigenetically upregulated by the histone acetyltransferase P300 and contributed to CD8⁺ T cell dysfunction and MEK/ERK pathway activation, thereby driving tumor progression. CONCLUSION: In summary, we established a novel fatty-acid-metabolism-related prognostic model for assessing the prognosis and potential immunotherapy response of NPC patients, as well as for characterizing the immunological features of the tumor microenvironment (TME). Furthermore, ABCC1 emerged as a promising prognostic biomarker associated with immunotherapeutic responsiveness in NPC, warranting further validation. CLINICAL TRIAL NUMBER: Not applicable.
Bovine lactoferrin induces apoptosis and modulates the transcriptomic landscape of HeLa cervical cancer cells.
Zhao Y, Liu R, Li M … +4 more , Feng M, Bao Y, Chen G, Zhang H
Biol Direct · 2026 Jun · PMID 42351212 · Full text
Cervical cancer (CC) remains a leading malignancy among women worldwide. Current management strategies, including surgery, radiotherapy, chemotherapy, targeted therapy, and prevention, often cause significant adverse eff...
Cervical cancer (CC) remains a leading malignancy among women worldwide. Current management strategies, including surgery, radiotherapy, chemotherapy, targeted therapy, and prevention, often cause significant adverse effects and demonstrate limited efficacy in advanced disease. Consequently, identifying novel therapeutic strategies and drug targets, particularly low-toxicity, high-specificity natural agents-constitutes a critical research priority. Bovine lactoferrin (bLF), a multifunctional iron-binding glycoprotein, demonstrates therapeutic potential, though its precise molecular mechanisms in cervical cancer are incompletely understood. This study therefore examined the biochemical and transcriptomic alterations in bLF-treated cervical cancer cells to elucidate its mode of action. bLF suppressed HeLa cell viability in a dose-dependent manner and concurrently promoted apoptosis while inhibiting proliferation and migration. The bLF-induced apoptosis appears to arise from a synergistic multi-mechanistic interplay, potentially involving increased intracellular ROS levels, downregulated mitochondrial membrane potential, or modulated expression of autophagy-related proteins to activate autophagic pathways. RNA sequencing identified substantial transcriptomic changes in cancer-associated pathways, notably affecting cell cycle progression, protein transport, apoptotic processes, and the negative regulation of apoptosis. These results substantiate the potent anticancer activity of bLF and highlight its promise as a therapeutic candidate derived from a natural product.
Programmed cell death-related gene S100A9 promotes macrophage M1 polarization and chondrocyte apoptosis in rheumatoid arthritis.
Xu Q, Liu J, Ding Q … +8 more , Zhao C, Wang W, Li H, Niu C, Chen W, Zeng P, Guan D, Zhang R
Biol Direct · 2026 Jun · PMID 42351193 · Full text
BACKGROUND: Rheumatoid arthritis (RA) is a heterogeneous chronic autoimmune disease. Its high disability rate has a serious impact on individuals and society. Programmed cell death (PCD) patterns play a key role in sever...
BACKGROUND: Rheumatoid arthritis (RA) is a heterogeneous chronic autoimmune disease. Its high disability rate has a serious impact on individuals and society. Programmed cell death (PCD) patterns play a key role in several diseases. However, the significance of the interplay between PCD and RA remains underexplored. METHODS: In total, 18 PCD patterns were analyzed for the model construction. Single-cell RNA-seq transcriptome (scRNA-seq) and bulk RNA-seq data were collected from the GSE200815, GSE1919, GSE77298, GSE206848, GSE89408, GSE12021, GSE55235, and GSE55457 cohorts to validate the model. In vivo and in vitro experiments were performed to determine the role of S100A9 in RA. RESULTS: We developed a programmed cell death-related (PCDR) model for RA using 113 combinations of 12 machine learning algorithms and significant PCD signatures; 2 RA clusters were identified. A significant difference was noted in the macrophage numbers between the two groups. Macrophages were identified as key effector cells that play a central role in RA pathogenesis through cellular communication and the transition of cell states. S100A9 was identified as a key gene in the PCDR model, and its knockdown significantly slowed RA progression by reducing joint synovitis and cartilage damage. M1 macrophage polarization was accompanied by the overexpression of S100A9 in the synovial tissues of RA model mice. Compared with RA mice, AAV-shRNA-mediated S100A9 knockdown mice showed decreased M1 macrophage polarization, attenuated severity of synovitis, and elevated expression of the cartilage phenotype proteins-collagen II and BCL-2. Additionally, S100A9 knockdown inhibited M1 macrophage polarization in vitro. Hence, S100A9 inhibition may be a promising therapeutic strategy for RA treatment. CONCLUSION: We established a novel PCDR model by comprehensively analyzing diverse cell death patterns. S100A9 inhibition may be a promising therapeutic strategy for RA treatment.
The role of ESRRA and JUNB in oxyphil cells: insights from spatial transcriptomics and single-cell transcriptomics in uremic secondary hyperparathyroidism.
Zhang Z, Qiu X, Sun B … +2 more , Li X, Xia F
Biol Direct · 2026 Jun · PMID 42343487 · Full text
Oxyphil cells are a primary cellular component of the parathyroid gland. Due to the limitations of techniques for separating oxyphil cells from chief cells, the characteristics and functions of oxyphil cells in uremic se...
Oxyphil cells are a primary cellular component of the parathyroid gland. Due to the limitations of techniques for separating oxyphil cells from chief cells, the characteristics and functions of oxyphil cells in uremic secondary hyperparathyroidism (SHPT) remain largely unclear. Therefore, we integrated spatial transcriptomics and single-cell transcriptomics to investigate the characteristics of oxyphil cells and the association between oxyphil cells and calcitriol resistance in SHPT. 6 uremic SHPT samples and 3 normal parathyroid samples were used for single-cell transcriptomics, while the most suitable SHPT sample was used for spatial transcriptomics. By integrating spatial transcriptomics with single-cell transcriptomics data (93073 cells), we identified a subset of 7653 cells with high confidence as oxyphil cells. Both transcriptomic data indicated a higher mitochondrial transcript proportion in oxyphil cells, and further investigation revealed that the transcription factor estrogen related receptor alpha (ESRRA), participated in mitochondrial biogenesis in these cells. Meanwhile, lower VDR expression and calcitriol resistance were observed in oxyphil cells. Further investigation demonstrated that the transcription factor JUNB promoted the transcription of VDR gene by binding to the promoter area of VDR gene, thus ameliorating calcitriol resistance of oxyphil cells in SHPT. These findings offer reliable evidence and new insights into the characteristics of oxyphil cells and their potential role in SHPT, and provide JUNB as a potential target for calcitriol resistance in SHPT.
Tenascin C promotes the formation of abdominal aortic aneurysm by regulating the homeostasis of vascular smooth muscle cells.
Xiao X, Xiong J, Bao Y … +11 more , Huang C, Chen G, Lu H, Yang C, Li M, Lu W, Song H, Lin B, Lai Y, Liang H, Zhong L
Biol Direct · 2026 Jun · PMID 42343484 · Full text
BACKGROUND: Abdominal aortic aneurysm (AAA) is the most common type of arterial dilation disease. Once ruptured and bleeding, AAA can be life-threatening, with a mortality rate as high as 90%. Vascular smooth muscle cell...
BACKGROUND: Abdominal aortic aneurysm (AAA) is the most common type of arterial dilation disease. Once ruptured and bleeding, AAA can be life-threatening, with a mortality rate as high as 90%. Vascular smooth muscle cells(VSMCs) dyshomeostasis is the main pathological factor for AAA formation, but the specific regulatory process remains unclear. METHODS: Firstly, we employed single-cell RNA sequencing (scRNA-seq) to reveal the heterogeneity and intercellular communication network of VSMCs in the aorta of AAA patients. We further integrated the data from the public database to clarify differences of the target signal in AAA, and also collected clinical data to analyze both differences and predictive performance for AAA. Subsequently, we generated VSMCs-specific knockout mice in the PPE-induced AAA mouse model to observe formation of AAA and VSMCs dyshomeostasis. Finally, we combined methods such as immunoprecipitation, bioinformatics, western blotting, qPCR, transwell co-culture and luciferase reporter assays to clarify the specific molecular mechanisms underlying VSMCs dyshomeostasis. RESULTS: Through multi-level analysis and experiments, we confirmed that the Tenascin C (TNC) communication signal was significantly highly expressed in the arterial tissue and plasma of AAA, and elevated TNC levels in plasma could predict the occurrence of human AAA. In mouse models, we found that VSMCs-specific TNC knockout could inhibit PPE-induced AAA formation and VSMCs dyshomeostasis. Mechanistically, we revealed that TNC promoted nuclear translocation of NF-κB p50 and p65 through TLR4, thereby causing VSMCs dyshomeostasis, while the endoplasmic reticulum stress factor CHOP could induce TNC production in VSMCs. CONCLUSION: Our research demonstrates the existence of a TNC-centered signaling axis mediating VSMCs dyshomeostasis in the formation of AAA.
Silymarin attenuates diabetic nephropathy in rats via modulation of the miRNA-223/NLRP3/caspase-1/GSDMD axis and inflammasome-related pyroptotic signaling.
Aboismaiel MG, Amin MN, Eissa LA
Biol Direct · 2026 Jun · PMID 42343466 · Full text
Diabetic nephropathy (DN) is a progressive microvascular complication of diabetes and a leading cause of end-stage renal disease, with limited disease-modifying therapeutic options. DN is characterized by progressive ren...
Diabetic nephropathy (DN) is a progressive microvascular complication of diabetes and a leading cause of end-stage renal disease, with limited disease-modifying therapeutic options. DN is characterized by progressive renal dysfunction, oxidative stress, inflammation, fibrosis, and activation of inflammasome-related pathways. Silymarin, a natural compound with antioxidant and anti-inflammatory properties, has demonstrated renoprotective potential; however, its association with miRNA-223/NLRP3/caspase-1/GSDMD signaling in DN remains incompletely defined. Therefore, the present study investigated the potential renoprotective effects of silymarin in a high-fat diet/streptozotocin (HFD/STZ)-induced rat model of DN, with particular focus on miRNA-223 expression and inflammasome-associated pyroptotic signaling. Diabetic rats received oral silymarin (100 mg/kg/day) either for 12 weeks as an early intervention or 4 weeks as a delayed treatment. Renal function indices, blood glucose, HbA1c, serum insulin, HOMA-IR, LDH, lipid profile, body weight, kidney weight/body weight index, oxidative stress markers, and histopathological changes were evaluated. Renal expression of miRNA-223, NOD-like receptor family pyrin domain-containing 3 (NLRP3), caspase-1, gasdermin D (GSDMD), nuclear factor-kappa B (NF-κB) (p65), hypoxia-inducible factor-1 alpha (HIF-1α), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and fibronectin was assessed using RT-qPCR, ELISA, and immunohistochemistry. Silymarin significantly improved renal function, glycemic control, insulin resistance, lipid profile, oxidative stress, inflammatory mediators, HIF-1α expression, LDH activity, histopathological injury, and fibrosis relative to untreated DN rats. Moreover, silymarin was associated with increased renal miRNA-223 expression and reduced expression of NLRP3, caspase-1, and GSDMD. These effects were more pronounced in the early intervention group compared with delayed treatment. In conclusion, silymarin attenuated experimental DN via modulation of the miRNA-223/NLRP3/caspase-1/GSDMD axis and suppression of pyroptosis-associated signaling, alongside improvement of oxidative stress, inflammation, hypoxia, fibrosis, and metabolic disturbances. Further mechanistic studies are warranted to clarify the causal relationship between miRNA-223 modulation and the observed renoprotective effects of silymarin.
Deciphering mitotic catastrophe-associated transcriptomic patterns in renal cell carcinoma: prognostic significance and immunotherapy-related associations.
Wang Z, Hu X, Lin Z … +3 more , Che F, Wei X, Zeng H
Biol Direct · 2026 Jun · PMID 42332836 · Full text
BACKGROUND: Renal cell carcinoma (RCC) is a highly heterogeneous malignancy with complex molecular features. Mitotic catastrophe (MC), a key regulator of cell fate during aberrant mitosis, contributes to tumor progressio...
BACKGROUND: Renal cell carcinoma (RCC) is a highly heterogeneous malignancy with complex molecular features. Mitotic catastrophe (MC), a key regulator of cell fate during aberrant mitosis, contributes to tumor progression and therapeutic response; however, its prognostic relevance in RCC remains unclear. METHODS: We integrated multi-omics, clinical, and imaging data from TCGA and multiple external cohorts. Prognostic mitotic catastrophe-related genes were identified to construct a mitotic catastrophe-related score (MCRS) using CoxBoost and random survival forest models. Associations with clinicopathologic features, immune contexture, and mutational profiles were analyzed. Single-cell RNA sequencing characterized Cyclin F (CCNF) expression and its association with immunotherapy-treated cohort outcomes, and these findings were further validated in vitro and in murine xenograft models. RESULTS: The nine-gene MCRG signature stratified patients into high- and low-risk groups with strong prognostic performance (TCGA 1-, 3-, 5-year AUCs > 0.9). High MCRS correlated with advanced stage, adverse CT features, angiogenesis, inflammatory signaling, regulatory T-cell infiltration, and elevated LAG3/TIGIT expression. MTOR mutations were enriched in high-risk tumors. SurvSHAP(t) analysis identified CCNF as the top prognostic contributor. CCNF was upregulated in tumor tissues and enriched in proliferating cells and epithelial cells. High CCNF expression was associated with worse survival and inferior outcomes in immunotherapy-treated cohorts. Overexpression of CCNF promoted RCC cell proliferation, migration, invasion, and in vivo tumor growth, partly through mTOR pathway activation. CONCLUSIONS: We established a robust MC-based prognostic model for RCC and identified CCNF as a key regulator of tumor progression and the immune microenvironment, characterized by increased Treg abundance and checkpoint expression. CCNF may represent a candidate biomarker and therapeutic target in RCC, although its relevance to outcomes in immunotherapy-treated cohorts requires further validation.
PCSK9 inhibitors do not increase cognitive risk.
Ni D, Feng Z, Liang D … +4 more , Qi Z, Lu Y, Zhu L, Li G
Biol Direct · 2026 Jun · PMID 42332816 · Full text
BACKGROUND: Proprotein convertase subtilisin-kexin type 9 (PCSK9) inhibitors are potent lipid-lowering therapies, however, concerns regarding their neurocognitive safety persist because of conflicting evidence. METHODS:...
BACKGROUND: Proprotein convertase subtilisin-kexin type 9 (PCSK9) inhibitors are potent lipid-lowering therapies, however, concerns regarding their neurocognitive safety persist because of conflicting evidence. METHODS: We applied an integrated approach combining real-world pharmacovigilance, genetic epidemiology, and preclinical models. Disproportionality analysis was performed using the FDA Adverse Event Reporting System (FAERS) data to assess cognitive adverse events. Two-sample Mendelian randomization (MR) was used to assess the causal effects of apolipoprotein B (ApoB) lowering and PCSK9 inhibition on Alzheimer's disease (AD) using UK Biobank and FinnGen data, with a factorial MR design to evaluate interactions stratified by ApoB and PCSK9 polygenic risk. Preclinically, hepatocyte-specific knockout and pharmacological inhibition of PCSK9 in AD-transgenic mice were assessed through cognitive testing and neuropathological assessment. RESULTS: FAERS analysis showed no increased reporting signal for cognitive adverse events with PCSK9 inhibitors. For AD, the ROR was below unity (ROR = 0.62, 95% CI 0.45-0.87), whereas no statistically significant signal was observed for dementia (ROR = 0.92, 95% CI 0.79-1.08). In contrast, statins showed significant disproportionality signals for AD (ROR = 2.86, 95% CI 2.51-3.26) and dementia (ROR = 1.51, 95% CI 1.36-1.67). MR analysis revealed no causal association between genetically proxied PCSK9 inhibition and AD risk (OR = 1.00, 95% CI 0.93-1.07), or between ApoB and AD risk (OR = 0.79, 95% CI 0.56-1.11). In AD-transgenic mice, PCSK9 knockout reduced low-density lipoprotein-cholesterol by 54% without impairing cognition or neuronal integrity, or increasing amyloid-β plaques and astrocytes. Pharmacological inhibition of PCSK9 similarly showed no cognitive deficits. CONCLUSIONS: Integrated evidence from pharmacovigilance, genetic epidemiology, and preclinical models robustly supports the neurocognitive safety of PCSK9 inhibitors, including in individuals with high ApoB levels. These findings affirm their long-term neurocognitive safety profile in the management of atherosclerotic cardiovascular disease.
HIF-1α/NREP axis promotes hypertrophic scar fibroblast proliferation and collagen deposition via activation of the TGF-β1/Smad pathway.
Chen B, Lin C, Wang X … +3 more , Zhu W, Zhang J, Chen L
Biol Direct · 2026 Jun · PMID 42332798 · Full text
BACKGROUND: Hypertrophic scar (HS) is a fibroproliferative skin disorder characterized by excessive fibroblast proliferation and collagen overproduction during aberrant wound healing. Although hypoxia is a well-recognize...
BACKGROUND: Hypertrophic scar (HS) is a fibroproliferative skin disorder characterized by excessive fibroblast proliferation and collagen overproduction during aberrant wound healing. Although hypoxia is a well-recognized driver of HS formation, the molecular mechanisms linking hypoxic signaling to fibroblast activation remain incompletely understood. METHODS: In this study, the expression of hypoxia-inducible factor 1α (HIF-1α), neuronal regeneration-related protein (NREP), and phosphorylated small mother against decapentaplegic family member 2/3 (p-Smad2/3) was examined in HS and matched normal tissues from patients. Hypertrophic scar fibroblasts (HSFBs) were cultured under hypoxic or normoxic conditions, followed by genetic manipulation of HIF-1α or NREP to assess their effects on fibroblast proliferation and collagen synthesis. The direct transcriptional regulation of NREP by HIF-1α was confirmed using dual-luciferase reporter and ChIP assays. RESULTS: Compared with normal tissues, HS tissues exhibited markedly elevated expression of HIF-1α, NREP, and phosphorylated Smad2/3. Hypoxia significantly increased HIF-1α and NREP levels in HSFBs, promoting fibroblast proliferation and collagen deposition, whereas knockdown of either HIF-1α or NREP abrogated these effects. Mechanistically, HIF-1α directly bound to the NREP promoter to enhance its transcription, and NREP overexpression partially rescued the suppressive effects of HIF-1α knockdown. Furthermore, HIF-1α-induced NREP upregulation activated the TGF-β1/Smad signaling cascade, while inhibition of this cascade counteracted the effect of NREP overexpression on the changes caused by HIF-1α silencing. CONCLUSION: Our findings identify a novel HIF-1α/NREP/TGF-β1-Smad axis that drives fibroblast hyperproliferation and excessive collagen accumulation in HS. Targeting this signaling pathway may offer a promising therapeutic strategy for preventing or treating HS formation.
Polystyrene nanoplastics induce size-associated oxidative-inflammatory stress, hepatic insulin-signaling impairment and metabolic disturbances in normal and diabetic mice.
Xu F, Jin Y, Sha Z … +4 more , Yang Y, Wang S, Chen Y, Yin J
Biol Direct · 2026 Jun · PMID 42316274 · Full text
BACKGROUND: Nanoplastics are emerging pollutants that can cross biological barriers and perturb metabolic homeostasis. Polystyrene nanoplastics (PS-NPs) are frequently detected in food and drinking water; however, their...
BACKGROUND: Nanoplastics are emerging pollutants that can cross biological barriers and perturb metabolic homeostasis. Polystyrene nanoplastics (PS-NPs) are frequently detected in food and drinking water; however, their size-dependent metabolic effects and the modifying role of pre-existing diabetes remain incompletely understood. This study aimed to investigate whether subchronic oral exposure to PS-NPs of different particle sizes altered hepatic insulin signaling and glucose-lipid homeostasis in normal and diabetic mice, and whether oxidative-inflammatory imbalance was associated with these responses. RESULTS: We found that PS-NP exposure induced size-dependent disturbances in metabolic and hepatic signaling endpoints. Among the tested particle sizes, 25 nm PS-NPs generally showed the largest apparent changes across several inflammatory, oxidative-stress, glycemic and insulin-signaling outcomes, although direct between-size differences were not uniformly established for all endpoints. PS-NP exposure increased serum levels of inflammatory cytokines (IL-6, CRP and TNF-α) and altered hepatic oxidative-stress markers, including the lipid peroxidation marker malondialdehyde (MDA) and antioxidant enzyme activities (CAT, SOD and GSH-Px), suggesting systemic inflammatory activation accompanied by hepatic oxidative-stress imbalance. These changes were associated with suppression of the hepatic IRS1-AKT-PPARγ axis and downregulation of GLUT4 expression, consistent with impaired insulin signaling and metabolic regulation. Compared with normal mice, diabetic mice exhibited exacerbated oxidative stress, inflammatory activation, and metabolic disturbance upon PS-NP exposure, suggesting higher susceptibility of metabolically compromised hosts to PS-NPs. Exploratory SEM provided supportive evidence for an association between oxidative-inflammatory imbalance and hepatic insulin-signaling impairment, while downstream links to systemic metabolic disturbance require further validation. Exploratory multivariate analyses, including principal component analysis (PCA), linear discriminant analysis (LDA), and random forest (RF), suggested response patterns associated with metabolic background and particle size, but these analyses should be interpreted as supportive pattern-recognition tools rather than validated predictive models. CONCLUSIONS: This study suggests that PS-NPs may act as metabolic stressors under the present experimental conditions, with their effects influenced by particle size and host metabolic background. By integrating classical toxicological methods with exploratory computational modeling, our findings provide mechanistic clues and preliminary data-driven evidence that PS-NP exposure may be associated with hepatic insulin-signaling impairment and metabolic disturbance, underscoring the need to assess nanomaterial safety in metabolically vulnerable populations.
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