Pub Date : 2025-11-01Epub Date: 2025-10-08DOI: 10.1016/j.yexcr.2025.114780
Shuwen Ge , Yandong Liu , Dean Li , Guojing Han , Kongyan Niu , Hujun Ju , Hao Zhou , Rui Liu , Zhengjiang Zhu , Nan Liu , Lefeng Qu
Nicotinamide mononucleotide (NMN), a precursory metabolite of NAD, has been demonstrated to boost cellular NAD level that is coupled with various age-related beneficial effects in animal models. NAD-capped RNA (NAD-RNA) represents a critical but poorly studied modification at the epitranscriptomic level. Here we examine the impact of NMN supplementation on NAD-RNA in human peripheral blood mononuclear cells (PBMCs). We demonstrated that NMN supplementation increases NAD turnover coupled with a reduction in NAD-capped RNAs in both human and dog, revealing blood-derived NAD-RNAs as potential biomarkers sensitized to NMN exposure.
{"title":"Epitranscriptomic analysis reveals features of NAD-capped RNAs upon supplementation of nicotinamide mononucleotide in human","authors":"Shuwen Ge , Yandong Liu , Dean Li , Guojing Han , Kongyan Niu , Hujun Ju , Hao Zhou , Rui Liu , Zhengjiang Zhu , Nan Liu , Lefeng Qu","doi":"10.1016/j.yexcr.2025.114780","DOIUrl":"10.1016/j.yexcr.2025.114780","url":null,"abstract":"<div><div>Nicotinamide mononucleotide (NMN), a precursory metabolite of NAD, has been demonstrated to boost cellular NAD level that is coupled with various age-related beneficial effects in animal models. NAD-capped RNA (NAD-RNA) represents a critical but poorly studied modification at the epitranscriptomic level. Here we examine the impact of NMN supplementation on NAD-RNA in human peripheral blood mononuclear cells (PBMCs). We demonstrated that NMN supplementation increases NAD turnover coupled with a reduction in NAD-capped RNAs in both human and dog, revealing blood-derived NAD-RNAs as potential biomarkers sensitized to NMN exposure.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"453 1","pages":"Article 114780"},"PeriodicalIF":3.5,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145274393","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-01Epub Date: 2025-10-14DOI: 10.1016/j.yexcr.2025.114797
Kai Li , Yaping Bai , Jingtong Wang , Li Ren , Anqi Mo , Haijun Liu , Wenjun Pei
Recent studies have highlighted the critical roles of long non-coding RNAs (lncRNAs) in tumorigenesis and progression. Here, we report that the lncRNA RNF216P1 is significantly upregulated in hepatocellular carcinoma (HCC) and contributes to tumor growth. To elucidate its underlying mechanisms, we first analyzed the transcriptional levels of RNF216P1 and its targets, miR-195-5p and autophagy related 4B cysteine peptidase (ATG4B), in HCC tissues using The Cancer Genome Atlas dataset, followed by validation with RT-qPCR. ATG4B protein levels were assessed by Western blotting. Functional assays—including xenograft models, CCK-8 viability tests, wound-healing assays, and Transwell migration assays—were performed to evaluate the role of RNF216P1 in HCC progression. Furthermore, the interactions between RNF216P1 and miR-195-5p, as well as between miR-195-5p and ATG4B, were confirmed by fluorescence in situ hybridization (FISH), RNA immunoprecipitation assays, and dual-luciferase reporter assays. Collectively, our findings demonstrate that RNF216P1 promotes malignant progression in HCC cells by acting as a competing endogenous RNA for miR-195-5p, thereby upregulating ATG4B and enhancing autophagy. This study identifies a novel ceRNA axis—RNF216P1/miR-195-5p/ATG4B—that plays a pivotal role in HCC development and may represent a potential therapeutic target.
{"title":"RNF216P1 functions as an oncogenic gene through modulating miR-195-5p/ATG4B axis in hepatocellular carcinoma","authors":"Kai Li , Yaping Bai , Jingtong Wang , Li Ren , Anqi Mo , Haijun Liu , Wenjun Pei","doi":"10.1016/j.yexcr.2025.114797","DOIUrl":"10.1016/j.yexcr.2025.114797","url":null,"abstract":"<div><div>Recent studies have highlighted the critical roles of long non-coding RNAs (lncRNAs) in tumorigenesis and progression. Here, we report that the lncRNA RNF216P1 is significantly upregulated in hepatocellular carcinoma (HCC) and contributes to tumor growth. To elucidate its underlying mechanisms, we first analyzed the transcriptional levels of RNF216P1 and its targets, miR-195-5p and autophagy related 4B cysteine peptidase (ATG4B), in HCC tissues using The Cancer Genome Atlas dataset, followed by validation with RT-qPCR. ATG4B protein levels were assessed by Western blotting. Functional assays—including xenograft models, CCK-8 viability tests, wound-healing assays, and Transwell migration assays—were performed to evaluate the role of RNF216P1 in HCC progression. Furthermore, the interactions between RNF216P1 and miR-195-5p, as well as between miR-195-5p and ATG4B, were confirmed by fluorescence in situ hybridization (FISH), RNA immunoprecipitation assays, and dual-luciferase reporter assays. Collectively, our findings demonstrate that RNF216P1 promotes malignant progression in HCC cells by acting as a competing endogenous RNA for miR-195-5p, thereby upregulating ATG4B and enhancing autophagy. This study identifies a novel ceRNA axis—RNF216P1/miR-195-5p/ATG4B—that plays a pivotal role in HCC development and may represent a potential therapeutic target.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"453 1","pages":"Article 114797"},"PeriodicalIF":3.5,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145307250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-09-03DOI: 10.1016/j.yexcr.2025.114711
Yanxu Chen , Qiang Zhang , Wenyu Xie , Pengfei Gao , Zhaowei Hu , Shenghui Wu , Zirong Bi , Huanxi Zhang , Yifang Gao , Changxi Wang , Longshan Liu
Background
Chronic rejection is a major cause of long-term kidney allograft failure, characterized by persistent inflammation and progressive fibrosis. Macrophages are central mediators of this process, but their phenotypic heterogeneity and regulatory mechanisms in chronic rejection remain incompletely understood.
Methods
We performed single-cell transcriptomic analysis on renal allograft biopsies from patients with different types of rejection and on a time-course rat model of chronic rejection. Macrophage subsets were identified through transcriptional profiling and Pseudotime trajectory analysis. Ligand–receptor analysis defined upstream intercellular communication, while in vitro assays using THP-1 macrophages evaluated responses to Jagged1 stimulation under polarizing conditions.
Results
A distinct TGFB+CD86+ macrophage subset exhibiting both pro-inflammatory and pro-fibrotic features was identified. This population, enriched in mixed rejection, occupied an intermediate position along the inferred macrophage trajectory and displayed dual ontogeny. It received Jagged1–NOTCH2 signals from mesenchymal-transitioned tubular epithelial cells and inflammatory inputs from infiltrating T cells. In vitro, co-stimulation with soluble Jagged1 under M1-polarizing conditions induced a similar hybrid phenotype. In the rat model, a phenotypically comparable subset, provisionally termed M2b, appeared early post-transplantation and was later replaced by M2a macrophages as fibrosis progressed. Ligand–receptor analysis confirmed conserved Jagged1–NOTCH2 signaling regulatory axis in vivo.
Conclusion
In summary, we identify a transitional TGFB+CD86+ macrophage population governed by JAG1–NOTCH2 signaling, bridging immune activation and fibrotic remodeling. Modulating this pathway may offer a therapeutic approach to reshape macrophage differentiation and mitigate chronic rejection.
{"title":"NOTCH signaling orchestrates the inflammatory-fibrotic continuum of macrophages in renal allograft rejection","authors":"Yanxu Chen , Qiang Zhang , Wenyu Xie , Pengfei Gao , Zhaowei Hu , Shenghui Wu , Zirong Bi , Huanxi Zhang , Yifang Gao , Changxi Wang , Longshan Liu","doi":"10.1016/j.yexcr.2025.114711","DOIUrl":"10.1016/j.yexcr.2025.114711","url":null,"abstract":"<div><h3>Background</h3><div>Chronic rejection is a major cause of long-term kidney allograft failure, characterized by persistent inflammation and progressive fibrosis. Macrophages are central mediators of this process, but their phenotypic heterogeneity and regulatory mechanisms in chronic rejection remain incompletely understood.</div></div><div><h3>Methods</h3><div>We performed single-cell transcriptomic analysis on renal allograft biopsies from patients with different types of rejection and on a time-course rat model of chronic rejection. Macrophage subsets were identified through transcriptional profiling and Pseudotime trajectory analysis. Ligand–receptor analysis defined upstream intercellular communication, while in vitro assays using THP-1 macrophages evaluated responses to Jagged1 stimulation under polarizing conditions.</div></div><div><h3>Results</h3><div>A distinct TGFB<sup>+</sup>CD86<sup>+</sup> macrophage subset exhibiting both pro-inflammatory and pro-fibrotic features was identified. This population, enriched in mixed rejection, occupied an intermediate position along the inferred macrophage trajectory and displayed dual ontogeny. It received Jagged1–NOTCH2 signals from mesenchymal-transitioned tubular epithelial cells and inflammatory inputs from infiltrating T cells. In vitro, co-stimulation with soluble Jagged1 under M1-polarizing conditions induced a similar hybrid phenotype. In the rat model, a phenotypically comparable subset, provisionally termed M2b, appeared early post-transplantation and was later replaced by M2a macrophages as fibrosis progressed. Ligand–receptor analysis confirmed conserved Jagged1–NOTCH2 signaling regulatory axis in vivo.</div></div><div><h3>Conclusion</h3><div>In summary, we identify a transitional TGFB<sup>+</sup>CD86<sup>+</sup> macrophage population governed by JAG1–NOTCH2 signaling, bridging immune activation and fibrotic remodeling. Modulating this pathway may offer a therapeutic approach to reshape macrophage differentiation and mitigate chronic rejection.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"452 2","pages":"Article 114711"},"PeriodicalIF":3.5,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145006032","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-09-19DOI: 10.1016/j.yexcr.2025.114767
Qing Zheng , Qingquan Tan , Dan Wang , Yanling Ma , Yanni Zhou , Yonghua Chen , Dan Long , Jiayin Yang , Li Feng
Liver fibrosis is a chronic progressive disease that can advance to cirrhosis or hepatocellular carcinoma if untreated. While liver transplantation remains the only curative option for end-stage fibrosis, the development of alternative therapies is urgently needed. In this study, we investigated the role of vitamin D-binding protein (VDBP) in hepatic fibrosis using clinical samples and a CCl4-induced mouse model. We observed significant downregulation of VDBP in fibrotic human and murine livers, suggesting that VDBP may serve as a potential biomarker for disease progression. VDBP knockout (VDBP-KO) mice exhibited exacerbated fibrosis, iron overload, and ferroptosis activation, accompanied by dysregulation of the Hippo-YAP pathway. In vitro, VDBP overexpression reversed these effects, while in vivo treatment with the YAP inhibitor verteporfin attenuated fibrosis, normalized iron homeostasis, and suppressed ferroptosis in VDBP-KO mice. Our findings demonstrate that VDBP plays a pivotal role in maintaining iron balance, inhibiting YAP signaling, and preventing ferroptosis during fibrogenesis. Elucidating the molecular mechanisms of VDBP and its downstream pathways may provide novel therapeutic targets for liver fibrosis. This could significantly improve the clinical management of hepatic fibrosis and offer new hope for patients suffering from this debilitating disease.
{"title":"Deficiency of vitamin D-binding protein exacerbates liver fibrosis by disrupting iron homeostasis via the activation of YAP signaling","authors":"Qing Zheng , Qingquan Tan , Dan Wang , Yanling Ma , Yanni Zhou , Yonghua Chen , Dan Long , Jiayin Yang , Li Feng","doi":"10.1016/j.yexcr.2025.114767","DOIUrl":"10.1016/j.yexcr.2025.114767","url":null,"abstract":"<div><div>Liver fibrosis is a chronic progressive disease that can advance to cirrhosis or hepatocellular carcinoma if untreated. While liver transplantation remains the only curative option for end-stage fibrosis, the development of alternative therapies is urgently needed. In this study, we investigated the role of vitamin D-binding protein (VDBP) in hepatic fibrosis using clinical samples and a CCl<sub>4</sub>-induced mouse model. We observed significant downregulation of VDBP in fibrotic human and murine livers, suggesting that VDBP may serve as a potential biomarker for disease progression. VDBP knockout (VDBP-KO) mice exhibited exacerbated fibrosis, iron overload, and ferroptosis activation, accompanied by dysregulation of the Hippo-YAP pathway. In vitro, VDBP overexpression reversed these effects, while in vivo treatment with the YAP inhibitor verteporfin attenuated fibrosis, normalized iron homeostasis, and suppressed ferroptosis in VDBP-KO mice. Our findings demonstrate that VDBP plays a pivotal role in maintaining iron balance, inhibiting YAP signaling, and preventing ferroptosis during fibrogenesis. Elucidating the molecular mechanisms of VDBP and its downstream pathways may provide novel therapeutic targets for liver fibrosis. This could significantly improve the clinical management of hepatic fibrosis and offer new hope for patients suffering from this debilitating disease.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"452 2","pages":"Article 114767"},"PeriodicalIF":3.5,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145109897","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-09-24DOI: 10.1016/j.yexcr.2025.114774
Renzhong Zhang, Xu Fu, Xiaoli Zhao, Ke Wang
Hyperuricemia (HUA)-induced renal injury involves elusive molecular mechanisms. This study uncovers suppressor of cytokine signaling 1 (SOCS1) as a pivotal mediator of hyperuricemic nephropathy through ferroptosis regulation. In a murine HUA model, we observed significantly elevated serum uric acid, impaired renal function, heightened inflammation, and activated ferroptosis. In vitro studies using uric acid-treated renal tubular cells demonstrated that SOCS1 deficiency alleviated ferroptotic cell death, reduced inflammatory responses, and preserved mitochondrial integrity. Mechanistically, SOCS1 directly interacts with glutathione peroxidase 4 (GPX4) to promote its ubiquitin-dependent proteasomal degradation, as validated by co-immunoprecipitation and protein stability assays. Crucially, pharmacological induction of ferroptosis abolished the protective effects of SOCS1 knockdown, while GPX4 inhibition counteracted its anti-ferroptotic function. In vivo delivery of renal-targeted SOCS1 shRNA via AAV9 vectors attenuated hyperuricemic nephropathy, ameliorating histological damage and suppressing both ferroptosis and inflammation. Our findings establish a pathogenic axis wherein SOCS1 drives hyperuricemic renal injury by facilitating GPX4 ubiquitination and subsequent ferroptosis activation, highlighting this pathway as a promising therapeutic target.
{"title":"SOCS1 orchestrates ferroptotic renal injury via GPX4 ubiquitination in hyperuricemia","authors":"Renzhong Zhang, Xu Fu, Xiaoli Zhao, Ke Wang","doi":"10.1016/j.yexcr.2025.114774","DOIUrl":"10.1016/j.yexcr.2025.114774","url":null,"abstract":"<div><div>Hyperuricemia (HUA)-induced renal injury involves elusive molecular mechanisms. This study uncovers suppressor of cytokine signaling 1 (SOCS1) as a pivotal mediator of hyperuricemic nephropathy through ferroptosis regulation. In a murine HUA model, we observed significantly elevated serum uric acid, impaired renal function, heightened inflammation, and activated ferroptosis. <em>In vitro</em> studies using uric acid-treated renal tubular cells demonstrated that SOCS1 deficiency alleviated ferroptotic cell death, reduced inflammatory responses, and preserved mitochondrial integrity. Mechanistically, SOCS1 directly interacts with glutathione peroxidase 4 (GPX4) to promote its ubiquitin-dependent proteasomal degradation, as validated by co-immunoprecipitation and protein stability assays. Crucially, pharmacological induction of ferroptosis abolished the protective effects of SOCS1 knockdown, while GPX4 inhibition counteracted its anti-ferroptotic function. <em>In vivo</em> delivery of renal-targeted SOCS1 shRNA <em>via</em> AAV9 vectors attenuated hyperuricemic nephropathy, ameliorating histological damage and suppressing both ferroptosis and inflammation. Our findings establish a pathogenic axis wherein SOCS1 drives hyperuricemic renal injury by facilitating GPX4 ubiquitination and subsequent ferroptosis activation, highlighting this pathway as a promising therapeutic target.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"452 2","pages":"Article 114774"},"PeriodicalIF":3.5,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145174398","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-09-25DOI: 10.1016/j.yexcr.2025.114773
Tadashi Sasagawa, Masabumi Shibuya
Preeclampsia (PE) is a major disease in the field of obstetrics. Onset and progression of PE are associated with abnormally high serum levels of soluble fms-like tyrosine kinase-1 (sFLT1), an anti-angiogenic factor primarily secreted by syncytiotrophoblasts (STBs) present in the placenta. Although a cell-based assay using primary human trophoblasts has been developed to identify compounds that inhibit sFLT1 secretion, routine application of this assay is limited owing to the complexity of isolating these cells from the placenta and their inability to be passaged. Recently, human trophoblast stem cell (hTSC) lines that can differentiate into STBs and extravillous trophoblasts have been established. Their high proliferative ability allows for obtaining sufficient STBs for drug screening. In the present study, we investigated whether hTSC-differentiated STBs (dSTBs) exhibit enhanced secretion of sFLT1 under hypoxic conditions, similar to primary trophoblasts. Hypoxic stimulation significantly increased sFLT1 secretion by the dSTBs. This response was markedly inhibited by small interfering RNAs targeting the hypoxia-inducible factor (HIF)-2α and HIF-1β, as well as by the HIF-2α inhibitor, belzutifan. These findings suggest that the dSTBs described above are a practical and scalable alternative to primary trophoblasts for drug screening in PE treatment.
{"title":"Human trophoblast stem cell-differentiated syncytiotrophoblasts as a model for hypoxia-enhanced secretion of the anti-angiogenic factor sFLT1","authors":"Tadashi Sasagawa, Masabumi Shibuya","doi":"10.1016/j.yexcr.2025.114773","DOIUrl":"10.1016/j.yexcr.2025.114773","url":null,"abstract":"<div><div>Preeclampsia (PE) is a major disease in the field of obstetrics. Onset and progression of PE are associated with abnormally high serum levels of soluble fms-like tyrosine kinase-1 (sFLT1), an anti-angiogenic factor primarily secreted by syncytiotrophoblasts (STBs) present in the placenta. Although a cell-based assay using primary human trophoblasts has been developed to identify compounds that inhibit sFLT1 secretion, routine application of this assay is limited owing to the complexity of isolating these cells from the placenta and their inability to be passaged. Recently, human trophoblast stem cell (hTSC) lines that can differentiate into STBs and extravillous trophoblasts have been established. Their high proliferative ability allows for obtaining sufficient STBs for drug screening. In the present study, we investigated whether hTSC-differentiated STBs (dSTBs) exhibit enhanced secretion of sFLT1 under hypoxic conditions, similar to primary trophoblasts. Hypoxic stimulation significantly increased sFLT1 secretion by the dSTBs. This response was markedly inhibited by small interfering RNAs targeting the hypoxia-inducible factor (HIF)-2α and HIF-1β, as well as by the HIF-2α inhibitor, belzutifan. These findings suggest that the dSTBs described above are a practical and scalable alternative to primary trophoblasts for drug screening in PE treatment.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"452 2","pages":"Article 114773"},"PeriodicalIF":3.5,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145174428","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-09-07DOI: 10.1016/j.yexcr.2025.114747
Danni Xu , Yu Fu , Huamao Sun , Yanda Lu , Bo Shen , Xinbao Hao
Background
Nasopharyngeal carcinoma (NPC) is a kind of tumor disease with high malignant degree. CREPT expression was elevated abnormally in multi-cancers. However, the role and regulatory mechanism of CREPT in NPC remains unknown.
Methods
NPC clinical samples, NPC cells, and nude mice served as experimental objects. The levels of molecules were detected using RT-qPCR,Western blot, IF and IHC experiments. Malignant activities of NPC cells were evaluated using CCK-8, EdU and clone formation, flow cytometry and TUNEL. Kaplan-Meier and Pearson correlation analysis were employed to analysis of the relationships between CREPT expression and prognosis/ALYREF expression in NPC patients. The interaction between ALYREF and CREPT was validated using RIP, RNA pull-down and dual luciferase experiment.
Results
Upregulation of CREPT and ALYREF expression was observed in NPC samples including the tissues of NPC patients and cells. CREPT knockdown reduced NPC cell viability, proliferation and enhanced NPC cell apoptosis and suppressed tumor growth through deactivating Wnt/β-catenin pathway, but the results of ALYREF overexpression had the inverse results of CREPT knockdown. Furthermore, the combination of CREPT knockdown and ALYREF overexpression compromised ALYREF overexpression-mediated the influences on NPC cells, tumor growth and motivating Wnt/β-catenin pathway. Furthermore, ALYREF interacted with CREPT mRNA and ALYREF promoted the stability of CREPT mRNA in m5C-dependent manner.
Conclusion
ALYREF stabilized CREPT mRNA through interacting with m5C-labeled CREPT mRNA to elevate CREPT expression, thus activating Wnt/β-catenin pathway and facilitating NPC progression.
{"title":"ALYREF stabilizes CREPT mRNA to accelerate the development of nasopharyngeal carcinoma through dependence on m5C modification","authors":"Danni Xu , Yu Fu , Huamao Sun , Yanda Lu , Bo Shen , Xinbao Hao","doi":"10.1016/j.yexcr.2025.114747","DOIUrl":"10.1016/j.yexcr.2025.114747","url":null,"abstract":"<div><h3>Background</h3><div>Nasopharyngeal carcinoma (NPC) is a kind of tumor disease with high malignant degree. CREPT expression was elevated abnormally in multi-cancers. However, the role and regulatory mechanism of CREPT in NPC remains unknown.</div></div><div><h3>Methods</h3><div>NPC clinical samples, NPC cells, and nude mice served as experimental objects. The levels of molecules were detected using RT-qPCR,Western blot, IF and IHC experiments. Malignant activities of NPC cells were evaluated using CCK-8, EdU and clone formation, flow cytometry and TUNEL. Kaplan-Meier and Pearson correlation analysis were employed to analysis of the relationships between CREPT expression and prognosis/ALYREF expression in NPC patients. The interaction between ALYREF and CREPT was validated using RIP, RNA pull-down and dual luciferase experiment.</div></div><div><h3>Results</h3><div>Upregulation of CREPT and ALYREF expression was observed in NPC samples including the tissues of NPC patients and cells. CREPT knockdown reduced NPC cell viability, proliferation and enhanced NPC cell apoptosis and suppressed tumor growth through deactivating Wnt/β-catenin pathway, but the results of ALYREF overexpression had the inverse results of CREPT knockdown. Furthermore, the combination of CREPT knockdown and ALYREF overexpression compromised ALYREF overexpression-mediated the influences on NPC cells, tumor growth and motivating Wnt/β-catenin pathway. Furthermore, ALYREF interacted with CREPT mRNA and ALYREF promoted the stability of CREPT mRNA in m5C-dependent manner.</div></div><div><h3>Conclusion</h3><div>ALYREF stabilized CREPT mRNA through interacting with m5C-labeled CREPT mRNA to elevate CREPT expression, thus activating Wnt/β-catenin pathway and facilitating NPC progression.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"452 2","pages":"Article 114747"},"PeriodicalIF":3.5,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145029123","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-09-15DOI: 10.1016/j.yexcr.2025.114753
Sen Li, Qingchuan Dong, Wei Ren, Yi Sun, Zhigang Wang, Liang Pan
Background
M2-polarized macrophages exhibit pro-tumorigenic functions in multiple malignancies, including prostate cancer (PC). Discs large homolog-associated protein 5 (DLGAP5) acts as an oncogenic driver in PC. This study aims to delineate the precise role and mechanistic basis of DLGAP5 in regulating PC-associated macrophage polarization.
Methods
Bioinformatics analyses were used to observe the expression profiling and polarization association of DLGAP5 and its upstream transcription factors (TFs). Cell phenotypes were assessed by measuring cell invasion, migration, apoptosis, and proliferation. After co-culture with PC cells, M2 macrophage polarization was evaluated by measuring M2-associated markers through quantitative PCR, Western blot, and flow cytometry. Xenograft tumor models were employed to validate the oncogenic role of DLGAP5 in vivo. Chromatin immunoprecipitation (ChIP) and luciferase reporter assays were used to confirmed the functional interaction between transcription factor AP-4 (TFAP4) and the DLGAP5 promoter.
Results
DLGAP5 and TFAP4 were upregulated in PC tumors and cell lines. Elevated DLGAP5 expression was significantly associated with higher Gleason score, advanced clinical stage, and worse prognosis. Depletion of DLGAP5 suppressed cell growth, migration, and invasion capacities in vitro. Moreover, DLGAP5 depletion attenuated the M2 polarization of PC-associated macrophages. Also, knockdown of DLGAP5 decreased tumorigenicity and macrophage M2 polarization in vivo. Mechanistically, TFAP4 transcriptionally activated the DLGAP5 promoter to upregulate DLGAP5 expression. Loss of TFAP4 suppressed tumor cell growth, migratory capacity, and invasiveness, as well as macrophage M2 polarization, all of which could be rescued through DLGAP5 re-expression. Additionally, TFAP4 activated the JAK2/STAT3 signaling through DLGAP5 in PC cells. Inhibition of JAK2/STAT3 signaling reversed DLGAP5-mediated malignant phenotypes and macrophage M2 polarization.
Conclusion
Our findings demonstrate that TFAP4-transcriptionally activated DLGAP5 drives PC progression by promoting tumorigenic properties and macrophage M2 polarization, establishing the TFAP4/DLGAP5 axis as a potential therapeutic target for PC.
{"title":"TFAP4/DLGAP5 promotes tumor progression and macrophage M2 polarization in prostate cancer by activating the JAK2/STAT3 signaling","authors":"Sen Li, Qingchuan Dong, Wei Ren, Yi Sun, Zhigang Wang, Liang Pan","doi":"10.1016/j.yexcr.2025.114753","DOIUrl":"10.1016/j.yexcr.2025.114753","url":null,"abstract":"<div><h3>Background</h3><div>M2-polarized macrophages exhibit pro-tumorigenic functions in multiple malignancies, including prostate cancer (PC). Discs large homolog-associated protein 5 (DLGAP5) acts as an oncogenic driver in PC. This study aims to delineate the precise role and mechanistic basis of DLGAP5 in regulating PC-associated macrophage polarization.</div></div><div><h3>Methods</h3><div>Bioinformatics analyses were used to observe the expression profiling and polarization association of DLGAP5 and its upstream transcription factors (TFs). Cell phenotypes were assessed by measuring cell invasion, migration, apoptosis, and proliferation. After co-culture with PC cells, M2 macrophage polarization was evaluated by measuring M2-associated markers through quantitative PCR, Western blot, and flow cytometry. Xenograft tumor models were employed to validate the oncogenic role of DLGAP5 <em>in vivo</em>. Chromatin immunoprecipitation (ChIP) and luciferase reporter assays were used to confirmed the functional interaction between transcription factor AP-4 (TFAP4) and the DLGAP5 promoter.</div></div><div><h3>Results</h3><div>DLGAP5 and TFAP4 were upregulated in PC tumors and cell lines. Elevated DLGAP5 expression was significantly associated with higher Gleason score, advanced clinical stage, and worse prognosis. Depletion of DLGAP5 suppressed cell growth, migration, and invasion capacities <em>in vitro</em>. Moreover, DLGAP5 depletion attenuated the M2 polarization of PC-associated macrophages. Also, knockdown of DLGAP5 decreased tumorigenicity and macrophage M2 polarization <em>in vivo</em>. Mechanistically, TFAP4 transcriptionally activated the DLGAP5 promoter to upregulate DLGAP5 expression. Loss of TFAP4 suppressed tumor cell growth, migratory capacity, and invasiveness, as well as macrophage M2 polarization, all of which could be rescued through DLGAP5 re-expression. Additionally, TFAP4 activated the JAK2/STAT3 signaling through DLGAP5 in PC cells. Inhibition of JAK2/STAT3 signaling reversed DLGAP5-mediated malignant phenotypes and macrophage M2 polarization.</div></div><div><h3>Conclusion</h3><div>Our findings demonstrate that TFAP4-transcriptionally activated DLGAP5 drives PC progression by promoting tumorigenic properties and macrophage M2 polarization, establishing the TFAP4/DLGAP5 axis as a potential therapeutic target for PC.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"452 2","pages":"Article 114753"},"PeriodicalIF":3.5,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145080096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bone marrow-derived mesenchymal stromal cells (BMSCs) are multipotent cells that have attracted considerable attention in regenerative medicine. Current in vitro test focus on biochemical assays of hair cell-like cells (HCLCs) derived from BMSCs associated with changes in electrophysiological properties. HCLCs were produced from BMSCs by culturing BMSCs with B27, EGF, FGF, and IGF-1. RNA Sequencing studies, immunocytochemistry (ICC) and double immunofluorescence staining were used to test hair cell-associated markers on day 17 and 21–26. Next, we performed whole-cell patch-clamp recording by utilizing current- and voltage-clamp techniques to assess changes in membrane potential and ionic currents during differentiation. Immunostaining assay reveals significant expression of myosin VIIA and SOX2 in cultured hair cells on day 21–26. We have also found 8 enhanced transcripts in differentiated cell genes (Wnt7a, Mgat5b, Myo7a, Pou4f3, SOX2, Atoh1, Map2k3, Actin) using RNA Sequencing. Electrophysiological results indicate that cells undergoing differentiation had an average resting membrane potential (RMP) of −11.93 ± 0.89 mV on day 17 and −58.96 ± 1.10 mV on days 21–26. Differentiated HCLCs displayed a mean resting membrane resistance of 171.66 ± 29.12 MΩ, membrane time constant of 10.73 ± 0.45 ms and membrane capacitance of 0.0625 ± 0.0087 pF, following 21–26 days in culture. Our results also showed cultured HCLCs express transcriptomic profile of this cell type. These findings indicate that alterations in RMP may serve as a valuable indicator for distinguishing HCLCs differentiation potential from BMSCs.
{"title":"A time course analysis of the electrophysiological and gene expression properties during differentiation of hair cell-like cells in culture","authors":"Ali Asghar Peyvandi , Shima Davoudi , Narges Bazgir , Mahyar Janahmadi , Hamid Norioun , Shahrokh Khoshsirat , Somayeh Niknazar","doi":"10.1016/j.yexcr.2025.114775","DOIUrl":"10.1016/j.yexcr.2025.114775","url":null,"abstract":"<div><div>Bone marrow-derived mesenchymal stromal cells (BMSCs) are multipotent cells that have attracted considerable attention in regenerative medicine. Current in vitro test focus on biochemical assays of hair cell-like cells (HCLCs) derived from BMSCs associated with changes in electrophysiological properties. HCLCs were produced from BMSCs by culturing BMSCs with B27, EGF, FGF, and IGF-1. RNA Sequencing studies, immunocytochemistry (ICC) and double immunofluorescence staining were used to test hair cell-associated markers on day 17 and 21–26. Next, we performed whole-cell patch-clamp recording by utilizing current- and voltage-clamp techniques to assess changes in membrane potential and ionic currents during differentiation. Immunostaining assay reveals significant expression of myosin VIIA and SOX2 in cultured hair cells on day 21–26. We have also found 8 enhanced transcripts in differentiated cell genes (Wnt7a, Mgat5b, Myo7a, Pou4f3, SOX2, Atoh1, Map2k3, Actin) using RNA Sequencing. Electrophysiological results indicate that cells undergoing differentiation had an average resting membrane potential (RMP) of −11.93 ± 0.89 mV on day 17 and −58.96 ± 1.10 mV on days 21–26. Differentiated HCLCs displayed a mean resting membrane resistance of 171.66 ± 29.12 MΩ, membrane time constant of 10.73 ± 0.45 ms and membrane capacitance of 0.0625 ± 0.0087 pF, following 21–26 days in culture. Our results also showed cultured HCLCs express transcriptomic profile of this cell type. These findings indicate that alterations in RMP may serve as a valuable indicator for distinguishing HCLCs differentiation potential from BMSCs.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"452 2","pages":"Article 114775"},"PeriodicalIF":3.5,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145185141","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-09-29DOI: 10.1016/j.yexcr.2025.114778
Harish C. Chandramoorthy , Raed Obaid Saleh , Viralkumar Mandaliya , Roopashree R , Hanen Mahmod Hulail , Subasini Uthirapathy , Renu Arya , Deepak Nathiya , Dina M.R. AlKhafaf
The NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3) inflammasome has recently been characterized as a major player in cancer-related inflammation, tumor progression, immune modulation, and metastasis. The role of extracellular vesicles (EVs), such as exosomes and microvesicles, as active carriers of NLRP3 signaling components is increasingly being recognized, as they modulate the tumor microenvironment as well as distant tissues primed for metastasis. This study will address the diverse roles of EV-mediated NLRP3 signaling in cancer immunopathogenesis, with a particular emphasis on its role in establishing a pre-metastatic niche and immune evasion. Tumor-derived EVs that include NLRP3 as cargo can reprogram stroma and immune cells at secondary sites in the body to support a metabolic role associated with metastatic colonization. While tumor-derived EVs promote extracellular matrix remodeling, angiogenesis, and the recruitment of immune-suppressive cells, this cascade of processes permits the development of a permissive niche for metastatic colonization, and simultaneously impairs anti-tumor immune surveillance. The provided EV-mediated crosstalk also enables tumor cells to evade immune detection through the downregulation of antigen-presenting cells and the activation of immune-suppressive pathways via NLRP3-dependent mechanisms. The molecular mechanisms underlying EV-driven inflammasome signaling could pave the way for the identification of a novel and specific biomarker, in which its modulation could potentially affect the immune system in the tumor microenvironment (TME) and participate in cancer immunopathogenesis. This review highlights the need to dissect further the interactions and context-specific roles of EV-NLRP3 across cancers to identify new therapeutic strategies for preventing metastasis and enhancing immune therapies.
{"title":"Extracellular-vesicle-mediated NLRP3 signaling in cancer","authors":"Harish C. Chandramoorthy , Raed Obaid Saleh , Viralkumar Mandaliya , Roopashree R , Hanen Mahmod Hulail , Subasini Uthirapathy , Renu Arya , Deepak Nathiya , Dina M.R. AlKhafaf","doi":"10.1016/j.yexcr.2025.114778","DOIUrl":"10.1016/j.yexcr.2025.114778","url":null,"abstract":"<div><div>The NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3) inflammasome has recently been characterized as a major player in cancer-related inflammation, tumor progression, immune modulation, and metastasis. The role of extracellular vesicles (EVs), such as exosomes and microvesicles, as active carriers of NLRP3 signaling components is increasingly being recognized, as they modulate the tumor microenvironment as well as distant tissues primed for metastasis. This study will address the diverse roles of EV-mediated NLRP3 signaling in cancer immunopathogenesis, with a particular emphasis on its role in establishing a pre-metastatic niche and immune evasion. Tumor-derived EVs that include NLRP3 as cargo can reprogram stroma and immune cells at secondary sites in the body to support a metabolic role associated with metastatic colonization. While tumor-derived EVs promote extracellular matrix remodeling, angiogenesis, and the recruitment of immune-suppressive cells, this cascade of processes permits the development of a permissive niche for metastatic colonization, and simultaneously impairs anti-tumor immune surveillance. The provided EV-mediated crosstalk also enables tumor cells to evade immune detection through the downregulation of antigen-presenting cells and the activation of immune-suppressive pathways via NLRP3-dependent mechanisms. The molecular mechanisms underlying EV-driven inflammasome signaling could pave the way for the identification of a novel and specific biomarker, in which its modulation could potentially affect the immune system in the tumor microenvironment (TME) and participate in cancer immunopathogenesis. This review highlights the need to dissect further the interactions and context-specific roles of EV-NLRP3 across cancers to identify new therapeutic strategies for preventing metastasis and enhancing immune therapies.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"452 2","pages":"Article 114778"},"PeriodicalIF":3.5,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145206082","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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