Applied Biochemistry and Biotechnology最新文献

{"title":"Metabolomic Profiling of Cow Urine of Various Breeds Reveals Bioactive Metabolites of Diverse Industrial Applications.","authors":"Jeetesh Kushwaha, Yashpal Singh, M S Mahesh, Sushil Kumar Yadav, Pratik N Sheth, Abhishek S Dhoble","doi":"10.1007/s12010-025-05300-6","DOIUrl":"https://doi.org/10.1007/s12010-025-05300-6","url":null,"abstract":"<p><p>Cow urine is widely utilized for medicinal and agricultural purposes in rural areas of India, with urine from indigenous cow breeds (Bos indicus) believed to offer unique benefits compared to that of exotic (Bos taurus) breeds. This research aimed to profile the metabolites present in the urine of indigenous breeds of cows using gas chromatography-mass spectrometry (GC-MS) and to explore the potential applications of the identified compounds by referencing the established literature. The various cow breeds included in the study were Gir, Sahiwal, Gangatiri, Hariana, Kankrej, Rathi, Gaolao, and Jersey. Cows employed in the study to collect the samples from various locations differed in their body weight, age, and stage of lactation. GC-MS analysis revealed a range of compounds, including ethanone, cresol, bis(2-ethylhexyl) phthalate, phenol, eicosane, pentanol, isobutyl ester, ethyl ester, binapacryl, trifluoroacetate, xylene, amylene hydrate, dibutyl ester, and formamide. Notably, several compounds were consistently observed across multiple indigenous breeds. For instance, bis(2-ethylhexyl) phthalate and xylene were found in nearly all indigenous breeds, while ethanone was detected in Gir, Sahiwal, Gangatiri, Kankrej, Hariana, Gaolao as well as Jersey cows. Similarly, eicosane and pentanol were present in Gangatiri and Hariana breeds. These overlapping chemical signatures highlight potential metabolic similarities among the studied cow breeds. The identified compounds are known for their diverse industrial and pharmaceutical applications, including use in disinfectants, flavorings, cosmetics, and agrochemicals as well as metabolic engineering. Thus, this study-for the first time-comprehensively delineated the comparative metabolite profile of cow urine among different breeds of cows. The spectrum of urinary metabolites identified could offer opportunities to foster bio-based innovations having multifarious applications, including new product developments, across diversified fields.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144666764","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanisms and Research Methods of Protein Modification in Virus Entry.","authors":"Yuyang Xiao, Mingyang Gao, Xianqi Mo, Jiamiao Lang, Zimeng Wang, Zhongjun Ma, Meng Yang, Bailu Tang, Dan Liu, Hailun He","doi":"10.1007/s12010-025-05333-x","DOIUrl":"https://doi.org/10.1007/s12010-025-05333-x","url":null,"abstract":"<p><p>Protein interactions are of paramount importance for the performance of biological functions within organisms. Post-translational modifications, including glycosylation and phosphorylation, regulate protein-protein interactions through non-covalent mechanisms. Glycosylation typically facilitates binding by altering surface properties, whereas phosphorylation can either enhance or disrupt interactions depending on context, collectively amplifying the biological impact of proteins. The entry of viruses and certain intracellular parasites into host cells is facilitated by these modifications, which permit the binding of ligands to receptors and the traversal of the cell membrane barrier. As research in this domain progresses, innovative methodologies are being developed, including protein microarrays and proximity-labeling techniques. These developments are being increasingly employed in disease prevention, therapeutics, and fundamental medical research. In light of the recent surge in emerging infectious diseases, the study of protein interactions has assumed heightened relevance. This review explores protein modifications, including glycosylation, phosphorylation, and ubiquitination, and focuses on their roles in viral entry. It highlights advanced methods for analyzing protein-protein interactions (PPIs), notably proximity labeling and protein microarrays, and concludes with novel insights into therapeutic development, aiming to inspire innovation in this evolving field.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144666763","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SMURF1 Affects Dental Implant Integration in Diabetes By Regulating PKG2 Expression.","authors":"Xue-Qin Wei, Sheng-Zhi Zhang","doi":"10.1007/s12010-025-05326-w","DOIUrl":"https://doi.org/10.1007/s12010-025-05326-w","url":null,"abstract":"<p><strong>Background: </strong>Poor dental implant has been reported in type II diabetes mellitus (T2DM), which seriously affects the patients' quality of life. The protein kinase type 2 (PKG2) is a target to regulate osteogenic differentiation. We aimed to investigate the effect and mechanisms of SMAD Specific E3 Ubiquitin Protein Ligase 1 (SMURF1, E3 ubiquitin ligase) on bone integration of dental implants in T2DM by regulating PKG2 in a ubiquitination-dependent manner.</p><p><strong>Methods: </strong>The dental implant model of diabetic rats and the high-glucose-induced bone marrow mesenchymal stem cell (BMSC) model were constructed. The changes in osteogenic differentiation indices were determined by immunohistochemical staining and alizarin red staining. The levels of insulin and glucose tolerance were assessed by glucose and insulin tolerance tests. Hematoxylin-eosin staining was performed to detect alveolar bone inflammation. PKG2, PERK, CHOP, SMURF1, p-PERK, GRP78, and ATF4 levels were examined via quantitative real-time PCR or western blot. The interaction between SMURF1 and PKG2 and PKG2 ubiquitination was analyzed by co-immunoprecipitation.</p><p><strong>Results: </strong>The T2DM model rats exhibited enhanced glucose tolerance and insulin resistance. PKG2 expression and Runx2 signal were reduced in T2DM rats following the dental implant. After treatment with high glucose in BMSCs, the expression of PKG2 and osteogenic differentiation were reduced. The proteasome inhibitor MG132 recovered PKG2 expression. The reduction of PKG2 was related to ubiquitination. SMURF1 is the key E3 ubiquitin ligase for PKG2. SMURF1 expression was increased in T2DM rats and BMSC cells induced by high glucose. Overexpression SMURF1 was downregulated the expression of PKG2 protein. SMURF1 regulated PKG2 expression via ubiquitination. PKG2 overexpression reversed the inhibition of osteogenic differentiation by SMURF1 overexpression, and inhibits the expression of endoplasmic reticulum stress related proteins. Additionally, these changes caused by SMURF1 overexpression were reversed by cinaciguat (PKG2 generator).</p><p><strong>Conclusions: </strong>In T2DM, SMURF1 regulated PKG2 expression through ubiquitination, thereby interfering with osteogenic differentiation and affecting the integration of dental transplantation.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144666765","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"USP7 Stabilizes USF1 to Aggravate ox-LDL-Induced Endothelial Injury Through the MYD88/NF-κB Pathway in Atherosclerosis.","authors":"Jing Liu, Xiangyang Zhang, Zhaoxia Yu, Tieliang Zhang","doi":"10.1007/s12010-025-05312-2","DOIUrl":"https://doi.org/10.1007/s12010-025-05312-2","url":null,"abstract":"<p><p>Atherosclerosis (AS) is a complex disease that involves the accumulation of lipids in the arterial wall, leading to vessel narrowing and increased risk of heart disease. Upstream stimulatory factor 1 (USF1) is an important regulatory factor that plays an important role in disease progression. Understanding the role and mechanism of USF1 in AS is crucial for unraveling the molecular underpinnings of this condition. Oxidized low-density lipoprotein (Ox-LDL) was used to stimulate human umbilical vein endothelial cells (HUVECs) to induce an AS-like cellular injury. Protein expression was evaluated using western blotting, while mRNA expression was assessed via quantitative real-time polymerase chain reaction. Cell viability and proliferation were analyzed using the cell counting kit-8 and 5-ethynyl-2'-deoxyuridine assays, respectively. Cell apoptosis was examined through flow cytometry. Angiogenic capacity was assessed by tube formation assay in human umbilical vein endothelial cells (HUVECs). Enzyme-linked immunosorbent assays were conducted to measure IL-6 and TNF-α levels, and MDA levels were determined using a lipid peroxidation MDA assay kit. SOD activity was measured using an SOD activity assay kit. Co-immunoprecipitation assay was performed to investigate the association between ubiquitin-specific peptidase 7 (USP7) and USF1, while dual-luciferase reporter assay and chromatin immunoprecipitation assay were conducted to identify the association between USF1 and myeloid differentiation primary response 88 (MYD88). USF1 expression was upregulated in AS patients when compared with healthy volunteers. Knockdown of USF1 protected HUVECs from injury induced by ox-LDL. USP7 was found to stabilize USF1 protein expression by deubiquitination, and its knockdown mitigated ox-LDL-induced HUVEC injury by reducing USF1 protein expression. USF1 was shown to transcriptionally activate MYD88 in HUVECs, and silencing of USF1 protected HUVECs from ox-LDL-induced injury by inhibiting MYD88 expression. Furthermore, USF1 knockdown inactivated the NF-κB pathway by suppressing MYD88 expression. USP7-dependent stabilization of USF1 exacerbated ox-LDL-induced injury in HUVECs by activating the MYD88/NF-κB pathway. These findings underscore the importance of the USP7-USF1-MYD88 axis in AS and suggest potential therapeutic targets for diseases related to AS.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144658099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Upregulated PCSK9 Level Contributes to Human Pulmonary Microvascular Endothelial Cell Apoptosis in Cigarette Smoke Extract-Induced COPD Models Through TLR4/NF-kappaB Pathway Activation.","authors":"Yunxia Li, Fengzhen He, Shasha Zhao, Chunli Zhang, Xueyang Chen, Xiang Luo","doi":"10.1007/s12010-025-05328-8","DOIUrl":"https://doi.org/10.1007/s12010-025-05328-8","url":null,"abstract":"<p><p>Chronic obstructive pulmonary disease (COPD) is persistent and can result in irreversible alveolar collapse and increase the risk of cardiovascular disease. PCSK9 belongs to the pro-protein convertase family and is capable of modulating cholesterol metabolism and low-density lipoprotein (LDL). PCSK9 is associated with endothelial dysfunction and its suppression is conducive to vascular function. The aim of this study was to explore PCSK9 expression in the lung tissues of patients with COPD and investigate the regulation of PCSK9 in cigarette smoke extract (CSE)-exposed human pulmonary microvascular endothelial cells (HPMECs) and an in vivo cigarette smoke (CS)-exposed mouse model. PCSK9 is drastically upregulated in the lung tissues of patients with COPD relative to those of individuals who have never smoked. PCSK9 expression in HPMECs and the CS-exposed mouse model was found to increase. Functional assays demonstrated that PCSK9 silencing decreased CS-induced lung injury and neutrophil and macrophage infiltration. PCSK9 silencing also abolished CSE-induced apoptosis by upregulating Bcl-2 and downregulating Bax expression in COPD mice and cell models. PCSK9 silencing alleviated the inflammatory response in the BALF of CS-exposed mice and CSE-treated HPMECs. The protein expression of P65, NLRP3, ASC, TLR4, p-P65, MyD88, and caspase-1 in mouse BALF and HPMECs was inhibited after PCSK9 knockdown. Collectively, these observations indicate the important role of PCSK9 in COPD progression and present promising treatment targets for COPD.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144658098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Astragalus Polysaccharides Alleviate Sepsis by Boosting Adrenal Hormone Synthesis and Repairing Microvascular Damage.","authors":"Yihui Chai, Yunzhi Chen, Zhaoxia Huang, Xiaoqing Liu, Jun Li","doi":"10.1007/s12010-025-05332-y","DOIUrl":"https://doi.org/10.1007/s12010-025-05332-y","url":null,"abstract":"<p><p>Sepsis is a severe infection-related condition commonly seen in clinical practice, characterized by systemic inflammation and multi-organ dysfunction. Astragalus polysaccharides (APS), the primary bioactive compound from the traditional Chinese herb Astragalus, has shown significant anti-inflammatory, immunomodulatory, and antioxidant effects. This study aimed to explore APS's potential in alleviating sepsis by enhancing adrenal cortical hormone production and repairing adrenal microvascular damage, while uncovering the underlying molecular mechanisms. Using an SD rat model, sepsis was induced via cecal ligation and puncture (CLP) surgery. Rats were treated with APS at doses of 400, 600, or 800 mg/kg/day for 7 days. Survival rates, inflammatory cytokine levels, adrenal function, and molecular pathways were evaluated using techniques such as ELISA, HE staining, TUNEL assay, metabolomics, transcriptome sequencing, and validation. Results showed that APS significantly reduced sepsis-induced mortality, elevated serum cortisol levels, and lowered inflammatory cytokines IL-6 and TNF-α. Histological analysis revealed that APS protected the adrenal cortex from apoptosis and inflammatory infiltration. Metabolomic analysis identified 154 differentially regulated metabolites, including significant changes in steroid hormones. Transcriptomic sequencing indicated changes in gene expression, suggesting that APS may inhibit NF-κB activity and promote nitric oxide (NO) production, thus protecting endothelial cells. These findings suggest that APS improves adrenal function and mitigates sepsis-related damage, offering promising therapeutic avenues for sepsis treatment.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144658097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Liposomal Delivery of Allolobophora caliginosa Coelomic Fluid Attenuates Myocardial Infarction by Suppressing Oxidative Damage, Inflammation, and Apoptosis.","authors":"Asmaa E Farouk, Sohair R Fahmy, Amel M Soliman, Sherif Abdelaziz Ibrahim, Shimaa A Sadek","doi":"10.1007/s12010-025-05340-y","DOIUrl":"https://doi.org/10.1007/s12010-025-05340-y","url":null,"abstract":"<p><p>Myocardial infarction (MI) is a concerning coronary heart disease with increasing rates of death and morbidity worldwide. One potential approach to prevent MI involves exploring invertebrate supplements within the nanoliposome formulation to improve targeted delivery, thereby mitigating MI-induced heart damage. Therefore, the study aimed to evaluate the cardioprotective efficacy of liposomal delivery of Allolobophora caliginosa coelomic fluid (ACCF-liposomes) on adrenaline-induced MI in rats. Thirty male albino rats were allocated into five groups: Control, Untreated MI, MI-treated ACCF, MI-treated free liposomes, and MI-treated ACCF-liposomes. The treatment regimen spanned 21 days. Electrocardiography (ECG), biochemical, oxidative stress, inflammatory mediators, electrolyte balance, histopathological and immunohistochemical analyses, and DNA fragmentation were evaluated. Liposomal delivery of ACCF has shown promise in regulating ECG criteria and reducing myocardial markers, particularly AST, LDH, MMP-2, creatine kinase, and troponin-I. It also improves lipid metabolism and inhibits myocardial oxidative stress. Additionally, ACCF and ACCF-liposomes treatment improves cardiomyocyte architecture and reduces DNA fragmentation in myocardial infarcted rats. Furthermore, encapsulating ACCF within liposomes statistically reduced the expression of iNOS and Beclin-1 in cardiac tissue. This suggests that liposomal delivery of ACCF enhances its effectiveness in treating myocardial infarction, potentially via its antioxidant, anti-inflammatory, and anti-apoptotic attributes.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641422","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hexanoic Acid Production from Chinese Cabbage Waste Driven by In Situ Lactic Acid Pre-Fermentation: Effect of pH.","authors":"Zhengang Chen, Liu Huang, Xiaofeng Ji, Ranran Chen, Jiying Zhu","doi":"10.1007/s12010-025-05334-w","DOIUrl":"https://doi.org/10.1007/s12010-025-05334-w","url":null,"abstract":"<p><p>Chain elongation (CE) with lactic acid as electron donor is an important way for the biosynthesis of hexanoic acid, and pH is crucial for the carbon flow direction in the CE process. In this study, a high concentration of lactic acid was achieved through the pre-fermentation of Chinese cabbage wastes (CCW), and the effects of pH on the metabolic flow of lactic acid and hexanoic acid yield during CE reaction were investigated. The coexistence of Lactobacillus, Caproiciproducens, and Acinetobacter at pH 5.5 facilitated the CE process driven by lactic acid, obtaining the highest hexanoic acid yield of 5.01 ± 0.01 g COD/L. At pH 6.0, the high abundance of propionic acid-producing bacteria, such as Blautia and Succiniclasticum, converted lactic acid to propionic acid via the acrylate pathway, resulting in low selectivity of hexanoic acid. Without pH control, acetic acid was the main product due to the low pH at the initial stage of fermentation.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641420","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improving the Accumulation and Solubility of Human Interferon-γ (hIFN-γ) in Escherichia coli: A Fusion Protein-Based Method and Network Pharmacology Analysis.","authors":"Narjes Akbari, Raheem Haddad, Reza Heidari-Japelaghi, Nematollah Gheibi","doi":"10.1007/s12010-025-05331-z","DOIUrl":"https://doi.org/10.1007/s12010-025-05331-z","url":null,"abstract":"<p><p>Using a network pharmacology-based method, some differentially expressed genes (DEGs) were detected in the colon cancer cell line (HCT116) after treatment with the human interferon-γ (hIFN-γ). Moreover, several pathways including cell cycle, NOD-like receptor signaling pathway, and P53 signaling pathway were identified, indicating the inhibitory effect of IFN-γ on the growth and proliferation of HCT116 cells. To validate in silico results, the hIFN-γ was first produced in Escherichia coli strain Rosetta and its bioactivity was then analyzed by anticancer assay. The production of hIFN-γ was performed via the optimization of several effective factors and the fusion of hIFN-γ to elastin like-polypeptide (ELP) tag. The highest amount of hIFN-γ (3.87 ± 0.37% of total soluble protein) was obtained at 22 °C with OD₆₀₀ = 0.6 and IPTG = 0.25 after 3-h inoculation. Whereas, the highest level of the hIFN-γ-ELP, about 4.58 ± 0.14% of TSP, was observed after 6-h inoculation. Compared to the hIFN-γ, the amount of hIFN-γ-ELP accumulation in the form of soluble increased significantly by more than 18%, proposing the desirable effect of ELP on the accumulation and solubility of hIFN-γ. Furthermore, the hIFN-γ prohibited the growth and proliferation of the HCT116 cells and the highest level of inhibition of cell proliferation was found at a concentration of 32.00 pg/mL hIFN-γ after 72-h incubation. Anticancer activity of hIFN-γ was also confirmed through the expression analysis of Bax, p53, and Bcl-2, suggesting the cytotoxic role of hIFN-γ toward HCT116 cells via inducing apoptosis process and arresting cell cycle.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TCF4 Promotes Neuroblastoma Proliferation and Inhibits Ferroptosis by Transactivating GPX4 Expression.","authors":"Yingming Wang, Qiang Gao, Xin Chen, Qian Dong, Ruihong Luan, Fujiang Li, Hongting Lu, Xianjun Zhou","doi":"10.1007/s12010-025-05329-7","DOIUrl":"https://doi.org/10.1007/s12010-025-05329-7","url":null,"abstract":"<p><p>Neuroblastoma, an aggressive pediatric malignancy, exhibits aberrant expression of transcription factors implicated in tumor progression. Here, we investigated the functional role of transcription factor 4 (TCF4) in neuroblastoma, focusing on its impact on cellular proliferation and ferroptosis-a regulated form of iron-dependent cell death, and elucidated the underlying molecular mechanism. Firstly, the expressions of TCF4 in neuroblastoma tissues and cell lines were analyzed, and the expressions of TCF4 mRNA and protein were significantly up-regulated. Functional analysis demonstrated that sh-TCF4 could significantly proliferate neuroblastoma cells, which was measured by CCK-8 kit, EdU staining, and clone formation experiments. Concurrently, TCF4 knockdown significantly elevated ROS accumulation and lipid peroxidation levels. Besides, sh-TCF4 decreased the levels of FTH1 and increased the TFR1 expression. Mechanistically, bioinformatic analysis using the JASPAR database predicted TCF4 binding sites within GPX4 promoter, a key ferroptosis regulator. ChIP and dual-luciferase reporter assays confirmed direct TCF4 occupancy and transcriptional activation of GPX4. Rescue experiments further validated the axis, as GPX4 overexpression abrogated the anti-proliferative and pro-ferroptotic effects induced by sh-TCF4. Collectively, the findings revealed TCF4 as a critical promoter of neuroblastoma growth and ferroptosis resistance, acting through direct up-regulation of GPX4. Targeting the TCF4-GPX4 axis may offer a novel therapeutic strategy to enhance ferroptosis sensitivity in neuroblastoma, warranting further preclinical exploration.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}