Journal of Biological Chemistry最新文献

{"title":"Microglial TAK1 promotes neurotoxic astrocytes and cognitive impairment in LPS-induced hippocampal neuroinflammation.","authors":"Xiao Han,Xin Cao,Qianqian Ju,Chengxin Ge,Yongqi Lin,Jinhong Shi,Xinhua Zhang,Cheng Sun,Haoming Li","doi":"10.1016/j.jbc.2025.110225","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110225","url":null,"abstract":"The peripheral immune system has a strong effect on the central nervous system (CNS). Systemic lipopolysaccharides (LPS) administration triggers robust microglial activation and induces significant inflammatory responses in the hippocampus. This study investigates the role of Transforming Growth Factor-β-Activated Kinase 1 (TAK1) in mediating LPS-induced hippocampal neuroinflammation and cognitive impairment. Our findings reveal that LPS induces activation of microglial TAK1, which in turn actives downstream effector NF-κB/p65 to release pro-inflammatory cytokines. The activated microglia also promote astrocytes to polarize into a neurotoxic phenotype (A1-like phenotype), and cause the loss of newborn neurons in the hippocampal dentate gyrus (DG). However, TAK1 reduction inhibits microglial responses, limits neurotoxic astrocytes, rescues newborn neurons, and subsequently improves LPS-induced cognitive deficits, suggesting that targeting TAK1 may be an effective strategy for alleviating neuroinflammation. The interaction between TAK1 activation, microglial responses, and the transition of neurotoxic astrocytes enhances our understanding of the cellular dynamics driving LPS-induced neuroinflammation, suggesting that TAK1 may be a therapeutic target for treating cognitive impairment.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"37 1","pages":"110225"},"PeriodicalIF":4.8,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932848","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inactivation of SIAH-1 E3 ligase attenuates Aβ toxicity by suppressing ubiquitin-dependent DVE-1 degradation in C. elegans models of Alzheimer's disease.","authors":"Lihua Sun,Jiahui Liu,Menghan Lu,Yingying Zhou,Shuqi Guo,Zhipeng Qin,Zekun Wang,Xiaojuan Sun","doi":"10.1016/j.jbc.2025.110226","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110226","url":null,"abstract":"The mitochondrial unfolded protein response (UPRmt), an evolutionarily conserved proteostasis pathway, plays a critical role in the pathogenesis of Alzheimer's disease (AD), characterized by amyloid-β peptide (Aβ) aggregation. Although the transcription factor DVE-1 regulates UPRmt activation in C. elegans and has been implicated in Aβ pathology, its regulatory mechanisms under AD-like conditions remain unclear. Here, using the classical C. elegans muscle-specific AD model (CL2006 strain), we observed UPRmt induction in young adults despite paradoxical depletion of DVE-1 protein concurrent with elevated dve-1 transcript levels. Through integrated genetic and biochemical analyses, we identified SIAH-1, a conserved E3 ubiquitin ligase that partners with the E2 enzyme UBC-25 to interact with DVE-1 and mediate its K48-linked polyubiquitination, as targeting DVE-1 for proteasomal degradation. Disruption of SIAH-1 E3 ubiquitin ligase function or overexpression of DVE-1 significantly reduced Aβ toxicity in both the muscle-expressed Aβ (CL2006) and neuronal Aβ models (gnaIs2). These interventions concurrently suppressed Aβ aggregation in the heat shock-inducible Aβ aggregation model (xchIs15). Mechanistically, this protective effect was associated with restored mitochondrial homeostasis, as evidenced by MitoTracker Red staining and TOMM-20::mCherry fluorescence imaging in muscle-expressed Aβ animals. These assays demonstrated that Aβ accumulation compromises mitochondrial integrity, a phenotype markedly rescued in siah-1 deletion mutants and DVE-1-overexpressing strains. Collectively, these findings establish the SIAH-1/DVE-1 axis as a conserved proteostasis regulator and highlight ubiquitin-dependent mitochondrial quality control as a potential therapeutic target for AD and related proteopathies.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"17 1","pages":"110226"},"PeriodicalIF":4.8,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering an antimicrobial chimeric endolysin that targets the phytopathogen Pseudomonas syringae pv. actinidiae.","authors":"Suzanne L Warring,Hazel M Sisson,George Randall,Dennis Grimon,Dorien Dams,Diana Gutiérrez,Matthias Fellner,Robert D Fagerlund,Yves Briers,Simon A Jackson,Peter C Fineran","doi":"10.1016/j.jbc.2025.110224","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110224","url":null,"abstract":"Global food shortages and rising antimicrobial resistance require alternatives to antibiotics and agrichemicals for the management of agricultural bacterial pathogens. The phytopathogen Pseudomonas syringae pv. actinidiae (Psa) is the causal agent of kiwifruit canker and is responsible for major agricultural losses. Bacteriophage enzymes present an emerging antimicrobial option. Endolysins possess the ability to cleave peptidoglycan and are effective antimicrobials against gram-positive bacteria. Delivery of endolysins to the peptidoglycan of gram-negatives is impeded by the additional outer membrane. To overcome this barrier, we used VersaTile molecular shuffling to produce Psa-targeting chimeric proteins which were tested for antimicrobial activity. These chimeras consist of endolysins linked by polypeptides to diverse phage proteins mined from Psa phage genomes. A preferential configuration for antibacterial activity was observed for enzymatic domains at the N-terminus and alternative phage proteins at the C-terminus. The lead variant possessed an N-terminal modular endolysin and a C-terminal lipase. Antibacterial activity was enhanced with the addition of the chemical permeabilizers citric acid or EDTA. Mutagenesis of the lipase active site eliminated exogenous antibacterial activity towards Psa. The endolysin-lipase chimera demonstrated specificity towards Psa, illustrating potential as a targeted biocontrol agent. Overall, we generated a chimeric endolysin with exogenous and specific activity towards Psa, the causative agent of kiwifruit canker.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"4 1","pages":"110224"},"PeriodicalIF":4.8,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"UCHL1 regulates adiponectin receptors in Sertoli cells to maintain testicular homeostatic balance.","authors":"Donghui Yang,Wenbo Chen,Ning Zhang,Mengfei Zhang,Wenping Wu,Lei Yang,Qizhong Lu,Shicheng Wan,Congliang Wang,Yuqi Wang,Xiaomin Du,Chao Huang,Zhengli Chen,Haiyang Tang,Na Li,Jinlian Hua","doi":"10.1016/j.jbc.2025.110221","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110221","url":null,"abstract":"Disruptions in testicular homeostasis can lead to impaired spermatogenesis and male infertility. Such disturbances may result from various factors, including viral or bacterial infections, toxic injuries, and genetic mutations or deletions. The maintenance of testicular homeostasis is governed by a complex interplay of various cells, hormones, paracrine factors, genes, and enzymes. UCHL1, a member of the deubiquitinating enzyme family, is recognized for its role in neuronal function. However, its contribution to testicular homeostasis and spermatogenesis remains unclear. This study uncovers a critical role for Uchl1 in maintaining testicular homeostasis, acting as a regulatory switch for spermatogenesis. We demonstrate that Uchl1 knockout (Uchl1_KO) mice exhibit reduced body weight, decreased testicular specific gravity, and impaired spermatogenesis. Single-nucleus RNA sequencing (snRNA-seq) analysis of Uchl1_KO testes reveals a significant decrease in oxidative phosphorylation (OXPHOS) levels and an increase in Sertoli cell abnormalities. Notably, Uchl1_KO/knockdown down-regulates metabolism-related adiponectin signaling (ADIPOR1/AMPK) and up-regulates the inflammation-related SEMA7A/PLXNC1 pathway. Sertoli cell lines (oeAdipor1/shUchl1) confirm UCHL1's dual regulatory role in these signaling pathways in vitro experiments. Our findings identify UCHL1 as a key regulator of testicular homeostasis and spermatogenesis, and it dynamically controls the balance between metabolic and inflammatory signaling in the testis. This study provides a valuable theoretical foundation for exploring the molecular mechanisms underlying testicular homeostasis balance and for advancing human reproductive health.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"10 1","pages":"110221"},"PeriodicalIF":4.8,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932851","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The macrophage sterol transport protein ORP2 promotes cholesterol efflux and prevents foam cell formation and atherosclerosis.","authors":"Xiaowei Wang,Kenan Peng,Yudi Zhao,Liwen Qiu,Chenxi Liang,Yaqian Dou,Qianqian Dong,Xiaoting Ma,Jinye Tang,Yidan Ma,Lin Liu,Mingqi Zheng,Hongyuan Yang,Mingming Gao","doi":"10.1016/j.jbc.2025.110228","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110228","url":null,"abstract":"Cholesterol-loaded macrophage foam cells are a key feature of atherosclerotic plaques. Oxysterol-binding protein-related protein 2 (ORP2) facilitates the transport of cholesterol from lysosomes to the plasma membrane in cultured cell lines. However, the role of ORP2 in macrophages and its involvement in atherosclerosis remain unclear. In this study, we found ORP2 expression was reduced in atherosclerotic vessels and in macrophages exposed to oxidized LDL (ox-LDL). Myeloid-specific human ORP2 overexpression (hORP2MOE) mice were generated and crossed with atherosclerotic-prone ApoE-/- mice, and then fed a high-fat diet (HFD) to induce atherosclerosis. Our results showed that myeloid-specific hORP2 overexpression significantly reduced the atherosclerotic plaque area, along with reduced lipid accumulation, necrotic core size, birefringent crystals, and macrophage presence within the plaque. Additionally, hORP2 overexpression in peritoneal macrophages (PMCs) let to reduced lipid accumulation and increased expression of key cholesterol efflux proteins, including LXRα, ABCA1, and ABCG1. Furthermore, hOPR2 overexpression promoted NBD-cholesterol efflux from macrophages. To explore the underlying mechanism, we conducted co-immunoprecipitation, immunofluorescence and cytoplasmic/nuclear fractionation experiments. Our findings revealed that ORP2 interacts with LXRα and promotes its nuclear localization in macrophages. Moreover, the LXR antagonist GSK2033 blocked the reduction in foam cell formation and the increase in LXRα nuclear translocation induced by hORP2 overexpression. These findings suggest that ORP2 interacts with LXRα and facilitates its nuclear translocation in macrophages, leading to reduced foam cell formation and alleviation of atherosclerosis.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"124 1","pages":"110228"},"PeriodicalIF":4.8,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932852","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Novel Compound DBZ Alleviates Chronic Inflammatory Pain and Anxiety-Like Behaviors by Targeting the JAK2-STAT3 Signaling Pathway.","authors":"Bao Sun,Mengyao Wu,Yilin Ru,Yaxi Meng,Xin Zhang,Fengyun Wang,Zhaodi Xia,Le Yang,Yufei Zhai,Gufeng Li,Jinming Hu,Bing Qi,Pu Jia,Sha Liao,Shixiang Wang,Minggao Zhao,Xiaohui Zheng","doi":"10.1016/j.jbc.2025.110223","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110223","url":null,"abstract":"Chronic pain profoundly disrupts patients' daily lives and places a heavy burden on their families. Tanshinol Borneol Ester (DBZ), a novel synthetic derivative, has demonstrated anti-inflammatory and anti-atherosclerotic effects, yet its impact on the central nervous system (CNS) remains largely unexplored. This study systematically examines the CNS effects of DBZ through a combination of in vivo, in vitro, network pharmacology, and molecular docking approaches. In vivo, we utilized a mouse model of chronic inflammation induced by complete Freund's adjuvant (CFA) to evaluate DBZ's influence on pain, anxiety-like behaviors, and its modulation of inflammatory and oxidative stress markers within the anterior cingulate cortex (ACC). In vitro studies on primary mouse astrocytes assessed DBZ's effects on cell viability and inflammatory marker expression. Network pharmacology was employed to elucidate DBZ's potential molecular targets and pathways, While molecular docking provides valuable docking confirmed its interactions with key components of the JAK2-STAT3 signaling pathway. Our findings demonstrate that DBZ effectively mitigates CFA-induced chronic pain and anxiety-like behaviors. It significantly suppresses astrocytes activation, reduces levels of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α, and diminishes oxidative stress markers such as ROS and MDA, while enhancing SOD levels. Moreover, DBZ modulates excitatory synaptic proteins and the JAK2-STAT3 signaling pathway in the ACC, suggesting its role in neuroprotection. These results position DBZ as a promising candidate for the treatment of chronic pain and anxiety, offering a potential foundation for the development of new therapeutic agents.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"21 1","pages":"110223"},"PeriodicalIF":4.8,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phosphorylation of SNX17 impedes activation of Retriever-mediated sorting.","authors":"Jan Dominik Speidel,Kaikai Yu,Ralph Thomas Böttcher","doi":"10.1016/j.jbc.2025.110222","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110222","url":null,"abstract":"Sorting nexin 17 (SNX17) functions as cargo receptor on endosomal membranes that enables the recycling of numerous membrane cargo proteins by binding to the Retriever complex. Yet, little is known how SNX17 activity or its membrane recruitment is regulated. Here, we report that phosphorylation of SNX17 at serine 38 (Ser38) within the phox (PX) domain serves as a critical regulatory switch governing its endosomal localization and function. A mutant form mimicking the phosphorylated state disrupts SNX17's ability to bind phosphatidylinositol-3-phosphate (PI3P), which in turn impairs its association with early endosomal membranes and inactivates SNX17-dependent cargo-recycling in cells. Furthermore, our results demonstrate that Ser38 is part of an autoinhibitory mechanism to regulate SNX17 cargo binding. Collectively, these findings provide new insights into the dynamic regulation of SNX17 activity and Retriever-mediated sorting processes. It also highlights SNX17 Ser38 phosphorylation as a critical regulatory mechanism that controls SNX17's endosomal localization and cargo recycling function.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"68 1","pages":"110222"},"PeriodicalIF":4.8,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The bacterial chaperone CsgC inhibits functional amyloid CsgA formation by promoting the intrinsically disordered pre-nuclear state.","authors":"Anthony Balistreri, Divya Kolli, Sanduni Wasana Jayaweera, Daniel Lundahl, Yilin Han, Lily Kalcec, Emily Goetzler, Rachel Alessio, Brandon Ruotolo, Anders Olofsson, Matthew R Chapman","doi":"10.1016/j.jbc.2025.110217","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110217","url":null,"abstract":"<p><p>E. coli assembles a functional amyloid called curli during biofilm formation. The major curlin subunit is the CsgA protein, which adopts a beta-sheet rich fold upon fibrillization. The chaperone protein CsgC inhibits CsgA amyloid formation. CsgA undergoes a 3-stage aggregation process: an initial lag phase where beta-rich nuclei form, an exponential elongation phase, and a plateau phase. It is currently not known if CsgC inhibits amyloid formation by inhibiting formation of a pre-fibril nucleus, or if CsgC inhibits a later stage of amyloid formation by blocking monomer addition. Here, CsgC homologs from C. youngae, C. davisae, and H. alvei were purified and characterized for their ability to interrogate CsgA amyloid formation. Each of the CsgC homologs prolonged the lag phase of E. coli CsgA amyloid formation similar to E. coli CsgC. Additionally, we found E. coli CsgC interacted transiently and weakly with a monomeric, pre-nucleus species of CsgA which delayed amyloid formation. A transient CsgC-CsgA heterodimer was observed using ion mobility-mass spectrometry. When CsgC was added to actively polymerizing CsgA, exponential growth commonly associated with nucleation-dependent amyloid formation was lost. Adding preformed CsgA seeds did not rescue exponential growth, indicating that CsgC also has inhibitory activity during fibril elongation. Indeed, CsgC interacted strongly with CsgA fibers, suggesting the interaction between CsgC and CsgA fibers can slow new fiber growth. CsgC displays unique inhibitory activity at multiple stages of amyloid formation and acts as an energy-independent chaperone that transiently interacts with prefibrillar CsgA and an amyloid fiber.</p>","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"110217"},"PeriodicalIF":4.0,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144010846","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acid sphingomyelinase recruits palmitoylated CD36 to membrane rafts and enhances lipid uptake.","authors":"Meng Ding,Yun Zhang,Xiaoting Xu,Yuan Zhu,Hui He,Tianyu Jiang,Yashuang Huang,Wenfeng Yu,Hailong Ou","doi":"10.1016/j.jbc.2025.110213","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110213","url":null,"abstract":"CD36 palmitoylation increases its membrane localization and is required for CD36-mediated uptake of oxidized low-density lipoprotein (oxLDL). Acid sphingomyelinase (ASMase) is transported to the plasma membrane, where it promotes lipid raft clustering, facilitating membrane protein anchoring for biological functions. We here investigated the effects of oxLDL on CD36 palmitoylation and explored the role of ASMase in CD36 membrane translocation. We found that oxLDL increased CD36 palmitoylation and drives its intracellular trafficking from the endoplasmic reticulum to plasma membrane lipid rafts in macrophages. Affinity purification followed by mass spectrometry analysis identified CD36 bound to ASMase in plasma membrane. The CD36/ASMase binding was enhanced by oxLDL treatment. Genetic ablation and pharmacological inhibition of ASMase reduced CD36 recruitment to lipid rafts, and inhibited CD36 intracellular signaling and lipid uptake. Moreover, inhibiting Sortilin to block ASMase intracellular trafficking and reduce membrane ASMase also caused a sharp decrease in amount of membrane CD36. In addition, ASMase overexpression dramatically promoted palmitoylated CD36 membrane localization but not CD36 without palmitoylation, in which the modification was inhibited by 2-bromopalmitate (2-BP) treatment or point mutation at the palmitoylation site. Moreover, ASMase knockout inhibited CD36 membrane recruitment both in peritoneal macrophages and in aorta, and attenuated lipid accumulation in atherosclerotic plaques in mice. Finally, we found oxLDL activated extracellular signal-regulated kinase1/2 (ERK1/2)/specificity protein (SP1) signaling, upregulating ASMase transcription and promoting sphingomyelin catabolism. Therefore, these data demonstrate that ASMase expression induced by oxLDL is required for palmitoylated CD36 membrane translocation during foam cell formation in macrophages.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"13 1","pages":"110213"},"PeriodicalIF":4.8,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932885","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the regulatory mechanism of CCNA2 in colorectal cancer: insights from multiomics and experimental analysis.","authors":"Xinyi Lei,Lanying Qiu,Qiang Chen,Lan Liao,Pengfei Yu,Wenjie Wu,Zhengyang Zhu,Chunying Li,Gang Lin,Zirui Zhuang,Yuxin Meng,Yan Wang,Cunchuan Wang,Yian Du","doi":"10.1016/j.jbc.2025.110216","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.110216","url":null,"abstract":"Colorectal cancer (CRC) is the third-most common cancer and the second-leading cause of mortality due to cancer worldwide. The underlying regulatory mechanism of CCNA2 in CRC was explored through multiomics and experimental analyses, thus facilitating diagnosis, therapy and prognosis. Two gene expression datasets (i.e., GSE9348 and GSE110223) were extracted from GEO. Differentially expressed genes (DEGs) were identified via GEO2R, which were used for enrichment analyses through DAVID. PPI network of DEGs was constructed by STRING, and the core genes were identified. CCNA2, a prognostic core gene for CRC, was validated in TCGA and HPA via transcriptomics and proteomics. ROC analysis was performed to evaluate the diagnostic value of CCNA2 in CRC. The therapeutic value of CCNA2 was evaluated in DGIdb through pharmacogenomics. The correlation between CCNA2 and immune infiltration was determined in TIMER by immunomics. TF-mRNA and miRNA-mRNA networks for CCNA2 were constructed in miRnet and miRDB via transcriptomics. The role and mechanism of CCNA2 in CRC were investigated both in vitro and in vivo. The miR-548x-3p/CCNA2 regulatory axis in CRC was investigated in vitro. CCNA2 showed excellent diagnostic, therapeutic, and prognostic value in CRC. CCNA2 was closely associated with tumor-infiltrating immunocytes, TFs, and miRNAs. The upregulation of CCNA2 was observed in CRC, and the knockdown of CCNA2 inhibited the proliferation, migration, and invasion while inducing apoptosis of CRC cells. The knockdown of CCNA2 could inhibit epithelial-mesenchymal transition (EMT) pathway. CCNA2 acted as a target of miR-548x-3p in regulating the biological behavior of CRC cells via the EMT-signaling pathway. CCNA2 is a potential biomarker for the diagnosis, treatment, and prognosis of CRC and is associated with immune infiltration, TF, and miRNA. The miR-548x-3p/CCNA2 axis plays a pivotal role in regulating the tumorigenesis of CRC through the EMT-signaling pathway.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"33 1","pages":"110216"},"PeriodicalIF":4.8,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}