Pub Date : 2025-11-19DOI: 10.1016/j.prp.2025.156300
Zhixing Zhang , Wei Kang , Chunjun Li , Dongdong Zhang , Xiaoyu Chen , Dan Lu , Yuzhen Huang , Lixia Zeng
Purpose
To investigate the presence and clinical significance of tertiary lymphoid structures (TLSs) in HER2-low breast cancer, by focusing on their associations with clinicopathological features and prognosis.
Methods
Hematoxylin and eosin staining and immunohistochemical markers were used in combination with whole-slide imaging (WSI) to delineate invasive carcinoma and adjacent TLSs. WSI-based tools were subsequently utilized to annotate the location, density, and maturity of TLSs.
Results
Among 560 patients with HER2-low breast cancer, the median age was 53 years and the age range was 28–85 years; 34 % (189/560) had TLSs-positive tumors. TLSs were associated with high histological grade, high-grade DCIS, absence of lymphovascular invasion, ER-negative, PR-negative, HER2 2 + , high K-i67, and triple-negative breast cancer subtypes (all P < 0.05). In the low-age cohort, TLSs (+), TLSs density, TLSs maturity, and peritumoral TLSs were significantly associated with poorer DFS (all P < 0.05).Conversely, TLSs (+), TLSs density, TLSs maturity, and peritumoral TLSs were significantly associated with better DFS in the high-age cohort (all P < 0.05).
Conclusion
In HER2-low breast cancer, TLSs were associated with higher histological grade, the presence of DCIS, absence of lymphovascular invasion, as well as expression of ER negative, PR negative, HER2 2 + , high K-i67, and triple-negative breast cancer. Additionally, the clinical prognosis value of TLSs (such as DFS) exhibited a correlation with the patient’s age.
{"title":"The clinical pathological significance of TLSs in HER2-low breast cancer","authors":"Zhixing Zhang , Wei Kang , Chunjun Li , Dongdong Zhang , Xiaoyu Chen , Dan Lu , Yuzhen Huang , Lixia Zeng","doi":"10.1016/j.prp.2025.156300","DOIUrl":"10.1016/j.prp.2025.156300","url":null,"abstract":"<div><h3>Purpose</h3><div>To investigate the presence and clinical significance of tertiary lymphoid structures (TLSs) in HER2-low breast cancer, by focusing on their associations with clinicopathological features and prognosis.</div></div><div><h3>Methods</h3><div>Hematoxylin and eosin staining and immunohistochemical markers were used in combination with whole-slide imaging (WSI) to delineate invasive carcinoma and adjacent TLSs. WSI-based tools were subsequently utilized to annotate the location, density, and maturity of TLSs.</div></div><div><h3>Results</h3><div>Among 560 patients with HER2-low breast cancer, the median age was 53 years and the age range was 28–85 years; 34 % (189/560) had TLSs-positive tumors. TLSs were associated with high histological grade, high-grade DCIS, absence of lymphovascular invasion, ER-negative, PR-negative, HER2 2 + , high K-i67, and triple-negative breast cancer subtypes (all P < 0.05). In the low-age cohort, TLSs (+), TLSs density, TLSs maturity, and peritumoral TLSs were significantly associated with poorer DFS (all P < 0.05).Conversely, TLSs (+), TLSs density, TLSs maturity, and peritumoral TLSs were significantly associated with better DFS in the high-age cohort (all P < 0.05).</div></div><div><h3>Conclusion</h3><div>In HER2-low breast cancer, TLSs were associated with higher histological grade, the presence of DCIS, absence of lymphovascular invasion, as well as expression of ER negative, PR negative, HER2 2 + , high K-i67, and triple-negative breast cancer. Additionally, the clinical prognosis value of TLSs (such as DFS) exhibited a correlation with the patient’s age.</div></div>","PeriodicalId":19916,"journal":{"name":"Pathology, research and practice","volume":"277 ","pages":"Article 156300"},"PeriodicalIF":3.2,"publicationDate":"2025-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145615957","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Vascular calcification (VC) is a pathological process characterized by the deposition of calcium phosphate crystals in blood vessels. Despite its clinical significance, the molecular mechanisms underlying VC remain poorly understood. This study integrated transcriptomic data from public datasets and experimental models to identify key regulators of VC. Human aortic smooth muscle cells (HASMCs) were induced to calcify using osteogenic medium (OM), followed by transcriptomic sequencing. Differential gene expression, functional enrichment, and machine learning-based hub gene identification, were performed. Experimental validation was conducted using in vitro and in vivo models. Transcriptomic analysis identified 278 differentially expressed genes (DEGs), 45 of which were associated with metabolism. Bioinformatic and machine learning approaches highlighted Osteomodulin (OMD), and Stanniocalcin 2 (STC2) as key regulators of VC. The iRegulon tool predicted that OMD and STC2 share a common transcription factor Activating Transcription Factor 4 (ATF4). In calcified human vascular tissues, ATF4, OMD, and STC2 expression levels were significantly upregulated, correlating with increased calcification markers such as RUNX2, ALP, and OCN. Functional studies demonstrated that ATF4 transcriptionally upregulates OMD and STC2 by binding to their promoter regions, then activated the PI3K/AKT signaling pathway, promoting osteogenic differentiation in HASMCs. In vivo experiments using AAV-SM22α-shATF4 confirmed that targeting ATF4 alleviates VC by suppressing OMD and STC2 expression and reducing calcium deposition. In conclusion, our study reveals that ATF4 promotes vascular calcification by transcriptionally upregulating OMD and STC2,which in turn activates the PI3K/AKT signaling pathway. These findings provides new evidence for the direct regulatory relationship between signaling nodes in the field of VC signaling network.
{"title":"ATF4 transcriptional regulation of OMD and STC2 drives vascular calcification progression via the PI3K/AKT pathway","authors":"Zhang Yue , Ming-Yan Wang , Chun-Ze Yuan , Jin-Wen Xu , Ke-Ke Shao","doi":"10.1016/j.prp.2025.156296","DOIUrl":"10.1016/j.prp.2025.156296","url":null,"abstract":"<div><div>Vascular calcification (VC) is a pathological process characterized by the deposition of calcium phosphate crystals in blood vessels. Despite its clinical significance, the molecular mechanisms underlying VC remain poorly understood. This study integrated transcriptomic data from public datasets and experimental models to identify key regulators of VC. Human aortic smooth muscle cells (HASMCs) were induced to calcify using osteogenic medium (OM), followed by transcriptomic sequencing. Differential gene expression, functional enrichment, and machine learning-based hub gene identification, were performed. Experimental validation was conducted using in vitro and in vivo models. Transcriptomic analysis identified 278 differentially expressed genes (DEGs), 45 of which were associated with metabolism. Bioinformatic and machine learning approaches highlighted Osteomodulin (OMD), and Stanniocalcin 2 (STC2) as key regulators of VC. The iRegulon tool predicted that OMD and STC2 share a common transcription factor Activating Transcription Factor 4 (ATF4). In calcified human vascular tissues, ATF4, OMD, and STC2 expression levels were significantly upregulated, correlating with increased calcification markers such as RUNX2, ALP, and OCN. Functional studies demonstrated that ATF4 transcriptionally upregulates OMD and STC2 by binding to their promoter regions, then activated the PI3K/AKT signaling pathway, promoting osteogenic differentiation in HASMCs. In vivo experiments using AAV-SM22α-shATF4 confirmed that targeting ATF4 alleviates VC by suppressing OMD and STC2 expression and reducing calcium deposition. In conclusion, our study reveals that ATF4 promotes vascular calcification by transcriptionally upregulating OMD and STC2,which in turn activates the PI3K/AKT signaling pathway. These findings provides new evidence for the direct regulatory relationship between signaling nodes in the field of VC signaling network.</div></div>","PeriodicalId":19916,"journal":{"name":"Pathology, research and practice","volume":"277 ","pages":"Article 156296"},"PeriodicalIF":3.2,"publicationDate":"2025-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145570136","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-17DOI: 10.1016/j.prp.2025.156299
Caihong Cao , Xing Feng
Objective
Breast cancer (BC) is the chief cause of malignancy-related deaths in women. This paper investigates how the EBF3-ACADL axis inhibits BC progression through Hippo/YAP signaling.
Methods
The differentially expressed gene ACADL in BC was screened by the bioinformatics databases, and ACADL expression in BC tumors and cells was verified by immunohistochemistry, RT-qPCR, and western blot analysis. BC cells were infected with an overexpressing-ACADL lentivirus to examine the effect of ACADL on BC cell growth. The in vivo impact of ACADL was investigated by constructing a xenograft tumor model. Cells were treated with XMU-MP-1, an inhibitor of MST1/2 kinase, to study the influence of ACADL on BC progression through the Hippo/YAP pathway. The upstream mechanism of abnormally elevated ACADL expression was analyzed by bioinformatics, and EBF3 expression in BC tumors and cells was verified. Regulatory assays were performed to confirm the binding relationship between ACADL and EBF3.
Results
ACADL and EBF3 were poorly expressed in BC tissues and cell lines. ACADL overexpression blocked p-YAP (Ser127), nuclear YAP localization, and canonical target genes (CTGF, CYR61, and ANKRD1), thereby suppressing BC cell growth and xenograft tumor development. XMU-MP-1 reversed the suppressive effect of ACADL overexpression on BC progression. EBF3 transcriptionally activated ACADL expression by binding to its promoter. EBF3 overexpression suppressed the malignant behavior of BC cells and xenograft tumor development in mice, which was reversed by ACADL knockdown.
Conclusion
EBF3 transcriptionally activates ACADL and blocks the Hippo/YAP pathway to inhibit BC progression.
{"title":"EBF3 transcriptionally activates ACADL to block the Hippo/YAP signaling pathway and inhibits breast cancer progression","authors":"Caihong Cao , Xing Feng","doi":"10.1016/j.prp.2025.156299","DOIUrl":"10.1016/j.prp.2025.156299","url":null,"abstract":"<div><h3>Objective</h3><div>Breast cancer (BC) is the chief cause of malignancy-related deaths in women. This paper investigates how the EBF3-ACADL axis inhibits BC progression through Hippo/YAP signaling.</div></div><div><h3>Methods</h3><div>The differentially expressed gene ACADL in BC was screened by the bioinformatics databases, and ACADL expression in BC tumors and cells was verified by immunohistochemistry, RT-qPCR, and western blot analysis. BC cells were infected with an overexpressing-ACADL lentivirus to examine the effect of ACADL on BC cell growth. The <em>in vivo</em> impact of ACADL was investigated by constructing a xenograft tumor model. Cells were treated with XMU-MP-1, an inhibitor of MST1/2 kinase, to study the influence of ACADL on BC progression through the Hippo/YAP pathway. The upstream mechanism of abnormally elevated ACADL expression was analyzed by bioinformatics, and EBF3 expression in BC tumors and cells was verified. Regulatory assays were performed to confirm the binding relationship between ACADL and EBF3.</div></div><div><h3>Results</h3><div>ACADL and EBF3 were poorly expressed in BC tissues and cell lines. ACADL overexpression blocked p-YAP (Ser127), nuclear YAP localization, and canonical target genes (CTGF, CYR61, and ANKRD1), thereby suppressing BC cell growth and xenograft tumor development. XMU-MP-1 reversed the suppressive effect of ACADL overexpression on BC progression. EBF3 transcriptionally activated ACADL expression by binding to its promoter. EBF3 overexpression suppressed the malignant behavior of BC cells and xenograft tumor development in mice, which was reversed by ACADL knockdown.</div></div><div><h3>Conclusion</h3><div>EBF3 transcriptionally activates ACADL and blocks the Hippo/YAP pathway to inhibit BC progression.</div></div>","PeriodicalId":19916,"journal":{"name":"Pathology, research and practice","volume":"277 ","pages":"Article 156299"},"PeriodicalIF":3.2,"publicationDate":"2025-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145570116","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-17DOI: 10.1016/j.prp.2025.156298
Longbao Yang , Fenrong Chen , Xiong Li, Xiaoke Sun, Hong Li, Haitao Shi, Gang Zhao
Objective
To investigate the intervention effect of epigallocatechin gallate (EGCG) on liver fibrosis and its underlying molecular mechanisms.
Methods
A mouse model of liver fibrosis induced by a high-fat diet was established, with groups divided into normal control, high-fat diet (HF) group, and HF + EGCG groups (low, medium, and high doses). The therapeutic effect of EGCG on liver fibrosis was evaluated by liver pathological scoring, detection of serum biochemical indicators, analysis of fibrotic markers, and Western blot for fibrotic protein expression. LX-2 cells were cultured in vitro and activated by TGF-β1. Molecular biology experiments (RT-qPCR, Western blot, immunofluorescence, co-immunoprecipitation, etc.) were used to explore the effects of EGCG on LX-2 cell activation, proliferation, migration, and its regulation of the TGF-β/Smad signaling pathway. Ubiquitination assays, molecular docking, and enzyme inhibitor interventions were performed to clarify the regulatory mechanism of EGCG on MUC5AC stability and its interaction with MDM2. Gene silencing/overexpression techniques were used to verify the critical role of the MDM2/MUC5AC axis in EGCG’s anti-fibrotic effect.
Results
In vivo experiments showed that EGCG dose-dependently improved liver histological damage in high-fat diet-fed mice, reduced serum levels of ALT, AST, and TBil, increased albumin and prothrombin time, decreased the expression of fibrotic markers such as hyaluronic acid (HA) and laminin (LN), and inhibited the expression of fibrotic proteins including α-SMA and collagen I. In vitro experiments confirmed that EGCG reduced activation, proliferation, and migration of LX-2 cells by inhibiting the TGF-β1/Smad signaling pathway (downregulating Smad2/3 phosphorylation and upregulating Smad7). Mechanistically, EGCG targeted and bound to MDM2, inhibiting MDM2-mediated ubiquitination and degradation of MUC5AC, thereby increasing MUC5AC protein stability. MUC5AC directly interacted with TGF-β1, further inhibiting the activation of the TGF-β1/Smad pathway. Additionally, overexpression of MDM2 reversed the upregulation of MUC5AC and the anti-fibrotic effect of EGCG, while supplementation of MUC5AC restored the intervention effect of EGCG, confirming that EGCG exerts its function through the MDM2/MUC5AC axis.
Conclusion
EGCG targets MDM2 to prevent MUC5AC from ubiquitination and degradation. The upregulated MUC5AC binds to TGF-β1 and inhibits the TGF-β1/Smad signaling pathway, thereby suppressing hepatic stellate cell activation and liver fibrosis progression. This study provides new potential targets and experimental basis for the prevention and treatment of liver fibrosis.
{"title":"EGCG inhibits hepatic stellate cell activity and liver fibrosis by targeting the MDM2/MUC5AC-mediated TGF-β1/Smad signaling pathway","authors":"Longbao Yang , Fenrong Chen , Xiong Li, Xiaoke Sun, Hong Li, Haitao Shi, Gang Zhao","doi":"10.1016/j.prp.2025.156298","DOIUrl":"10.1016/j.prp.2025.156298","url":null,"abstract":"<div><h3>Objective</h3><div>To investigate the intervention effect of epigallocatechin gallate (EGCG) on liver fibrosis and its underlying molecular mechanisms.</div></div><div><h3>Methods</h3><div>A mouse model of liver fibrosis induced by a high-fat diet was established, with groups divided into normal control, high-fat diet (HF) group, and HF + EGCG groups (low, medium, and high doses). The therapeutic effect of EGCG on liver fibrosis was evaluated by liver pathological scoring, detection of serum biochemical indicators, analysis of fibrotic markers, and Western blot for fibrotic protein expression. LX-2 cells were cultured in vitro and activated by TGF-β1. Molecular biology experiments (RT-qPCR, Western blot, immunofluorescence, co-immunoprecipitation, etc.) were used to explore the effects of EGCG on LX-2 cell activation, proliferation, migration, and its regulation of the TGF-β/Smad signaling pathway. Ubiquitination assays, molecular docking, and enzyme inhibitor interventions were performed to clarify the regulatory mechanism of EGCG on MUC5AC stability and its interaction with MDM2. Gene silencing/overexpression techniques were used to verify the critical role of the MDM2/MUC5AC axis in EGCG’s anti-fibrotic effect.</div></div><div><h3>Results</h3><div>In vivo experiments showed that EGCG dose-dependently improved liver histological damage in high-fat diet-fed mice, reduced serum levels of ALT, AST, and TBil, increased albumin and prothrombin time, decreased the expression of fibrotic markers such as hyaluronic acid (HA) and laminin (LN), and inhibited the expression of fibrotic proteins including α-SMA and collagen I. In vitro experiments confirmed that EGCG reduced activation, proliferation, and migration of LX-2 cells by inhibiting the TGF-β1/Smad signaling pathway (downregulating Smad2/3 phosphorylation and upregulating Smad7). Mechanistically, EGCG targeted and bound to MDM2, inhibiting MDM2-mediated ubiquitination and degradation of MUC5AC, thereby increasing MUC5AC protein stability. MUC5AC directly interacted with TGF-β1, further inhibiting the activation of the TGF-β1/Smad pathway. Additionally, overexpression of MDM2 reversed the upregulation of MUC5AC and the anti-fibrotic effect of EGCG, while supplementation of MUC5AC restored the intervention effect of EGCG, confirming that EGCG exerts its function through the MDM2/MUC5AC axis.</div></div><div><h3>Conclusion</h3><div>EGCG targets MDM2 to prevent MUC5AC from ubiquitination and degradation. The upregulated MUC5AC binds to TGF-β1 and inhibits the TGF-β1/Smad signaling pathway, thereby suppressing hepatic stellate cell activation and liver fibrosis progression. This study provides new potential targets and experimental basis for the prevention and treatment of liver fibrosis.</div></div>","PeriodicalId":19916,"journal":{"name":"Pathology, research and practice","volume":"277 ","pages":"Article 156298"},"PeriodicalIF":3.2,"publicationDate":"2025-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145605549","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-13DOI: 10.1016/j.prp.2025.156297
Lijuan Yang, Dongli Wang, Nan Yu, Caixia Zhu
F-box and WD repeat domain-containing 7 (FBXW7), a ubiquitinating enzyme, has been verified as a key factor linking to the mechanical overloading and chondrocyte senescence in the pathology of osteoarthritis (OA). Given the lack of deeply mechanism research on the regulation of OA by FBXW7, elucidation of the action mechanism of FBXW7 in OA could provide theoretical basis for the treatment of OA. OA model was established by injuring the anterior cruciate ligament (ACL). Ferrostatin-1 (Fer-1) was applied for analysis of ferroptosis. After overexpressed or silence of FBXW7, cell viability and apoptosis were determined via CCK-8 and TUNEL staining. The intracellular Fe2 + , GSH concentration, ROS levels and mitochondrial membrane potential were assessed by iron determination kit, ELISA, C11-BODIPY/DCFH-DA and JC-1 staining methods. Western blot and RT-qPCR were carried out for determination of ferroptosis-correlated factors (SLC7A11 and GPX4) and ECM-related factors (collagen II (Col II) and ADAMTS5). The interaction between SLC7A11 protein and FBXW7 was detected by immunofluorescence (IF) and immunoprecipitation (IP). Up-regulation of FBXW7, and down-regulation of SLC7A11 and GPX4 were observed in OA groups, compared to that in Control group. Moreover, FBXW7 overexpression significantly hindered cell viability, injured cell morphology, promoted apoptosis and reduced Col II protein level, while Fer-1 treatment blocked the function of FBXW7 overexpression in OA injury. Additionally, silence of FBXW7 showcased the opposite results, meanwhile decreased Fe2+ level, increased GSH release, reduced ROS content, raised mitochondrial membrane potential and elevated SLC7A11 and GPX4 in OA chondrocytes. Furthermore, SLC7A11 and FBXW7 were co-localized in chondrocytes and exhibited protein interaction. The ubiquitination degradation of SLC7A11 was accelerated by FBXW7 in chondrocytes, which was intercepted by MG132 treatment. In vivo experimental results further uncovered the alleviated functions of FBXW7 knockdown in ferroptosis and cartilage damage in OA model. The finding demonstrated that FBXW7 aggravated OA injury and ferroptosis, which might be linked to the ubiquitination degradation of SLC7A11.
F-box和WD重复结构域7 （FBXW7）是一种泛素化酶，已被证实是骨关节炎（OA）病理中机械超载和软骨细胞衰老的关键因素。鉴于FBXW7对OA的调控机制缺乏深入的研究，阐明FBXW7在OA中的作用机制可以为OA的治疗提供理论依据。通过损伤前交叉韧带（ACL）建立骨关节炎模型。应用铁抑素-1 （fer1）分析铁下垂。FBXW7过表达或沉默后，通过CCK-8和TUNEL染色检测细胞活力和凋亡情况。采用铁测定试剂盒、ELISA、C11-BODIPY/DCFH-DA和JC-1染色法检测细胞内Fe2 +、GSH浓度、ROS水平和线粒体膜电位。Western blot和RT-qPCR检测凋亡相关因子（SLC7A11和GPX4）和ecm相关因子（collagen II （Col II）和ADAMTS5）。采用免疫荧光（IF）和免疫沉淀（IP）检测SLC7A11蛋白与FBXW7的相互作用。与对照组相比，OA组FBXW7表达上调，SLC7A11和GPX4表达下调。FBXW7过表达显著抑制细胞活力，损伤细胞形态，促进细胞凋亡，降低Col II蛋白水平，而fe -1处理可阻断FBXW7过表达在OA损伤中的作用。此外，FBXW7沉默显示相反的结果，同时OA软骨细胞中Fe2+水平降低，GSH释放增加，ROS含量降低，线粒体膜电位升高，SLC7A11和GPX4升高。此外，SLC7A11和FBXW7在软骨细胞中共定位，并表现出蛋白质相互作用。软骨细胞中的FBXW7加速了SLC7A11的泛素化降解，MG132阻断了FBXW7。体内实验结果进一步揭示了FBXW7敲低对OA模型铁下垂和软骨损伤的缓解作用。研究结果表明，FBXW7加重OA损伤和铁下沉，这可能与SLC7A11的泛素化降解有关。
{"title":"FBXW7 promotes osteoarthritis injury by regulating SLC7A11 ubiquitination degradation and chondrocyte ferroptosis","authors":"Lijuan Yang, Dongli Wang, Nan Yu, Caixia Zhu","doi":"10.1016/j.prp.2025.156297","DOIUrl":"10.1016/j.prp.2025.156297","url":null,"abstract":"<div><div>F-box and WD repeat domain-containing 7 (FBXW7), a ubiquitinating enzyme, has been verified as a key factor linking to the mechanical overloading and chondrocyte senescence in the pathology of osteoarthritis (OA). Given the lack of deeply mechanism research on the regulation of OA by FBXW7, elucidation of the action mechanism of FBXW7 in OA could provide theoretical basis for the treatment of OA. OA model was established by injuring the anterior cruciate ligament (ACL). Ferrostatin-1 (Fer-1) was applied for analysis of ferroptosis. After overexpressed or silence of FBXW7, cell viability and apoptosis were determined via CCK-8 and TUNEL staining. The intracellular Fe<sup>2 +</sup> , GSH concentration, ROS levels and mitochondrial membrane potential were assessed by iron determination kit, ELISA, C11-BODIPY/DCFH-DA and JC-1 staining methods. Western blot and RT-qPCR were carried out for determination of ferroptosis-correlated factors (SLC7A11 and GPX4) and ECM-related factors (collagen II (Col II) and ADAMTS5). The interaction between SLC7A11 protein and FBXW7 was detected by immunofluorescence (IF) and immunoprecipitation (IP). Up-regulation of FBXW7, and down-regulation of SLC7A11 and GPX4 were observed in OA groups, compared to that in Control group. Moreover, FBXW7 overexpression significantly hindered cell viability, injured cell morphology, promoted apoptosis and reduced Col II protein level, while Fer-1 treatment blocked the function of FBXW7 overexpression in OA injury. Additionally, silence of FBXW7 showcased the opposite results, meanwhile decreased Fe<sup>2+</sup> level, increased GSH release, reduced ROS content, raised mitochondrial membrane potential and elevated SLC7A11 and GPX4 in OA chondrocytes. Furthermore, SLC7A11 and FBXW7 were co-localized in chondrocytes and exhibited protein interaction. The ubiquitination degradation of SLC7A11 was accelerated by FBXW7 in chondrocytes, which was intercepted by MG132 treatment. <em>In vivo</em> experimental results further uncovered the alleviated functions of FBXW7 knockdown in ferroptosis and cartilage damage in OA model. The finding demonstrated that FBXW7 aggravated OA injury and ferroptosis, which might be linked to the ubiquitination degradation of SLC7A11.</div></div>","PeriodicalId":19916,"journal":{"name":"Pathology, research and practice","volume":"277 ","pages":"Article 156297"},"PeriodicalIF":3.2,"publicationDate":"2025-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145570118","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-10DOI: 10.1016/j.prp.2025.156287
Yanping Zhang , Shazhou Ye , Suying Wang , Qi Ding , Jing Jin , Ming Zhao
Bladder cancer (BCa) is the most common malignancy of the urinary system. Despite advancements in novel targeted therapies and immunotherapy, the majority of patients remain incurable, and disease progression frequently occurs after treatment. Therefore, identifying new therapeutic strategies is crucial. Fatty acids are essential components of cell structure, playing roles in energy storage and serving as signaling molecules. In tumor tissues, due to abnormal blood vessel development, cancer cells primarily rely on de novo fatty acid synthesis to meet the demands of growth and proliferation. Stearoyl-CoA desaturase 1 (SCD1) is a key enzyme, widely recognized as a potential therapeutic target in various cancers. SCD1 promotes the synthesis of cell membranes by converting saturated fatty acids into monounsaturated fatty acids, thus supporting tumor cell growth. In this study, we conducted bioinformatics analysis using public datasets (including bulk RNA-seq and single-cell RNA-seq) and immunohistochemical examination of BCa tissues. Our findings reveal that SCD1 is specifically expressed in BCa cells and is associated with poor tumor grade and prognosis. Furthermore, drug sensitivity predictions and validations suggest that SCD1 enhances the sensitivity of BCa cells to trametinib. Therefore, SCD1 offers a promising new avenue for the early diagnosis, prognostic assessment, and optimization of personalized treatment strategies for BCa.
{"title":"SCD1 drives bladder cancer progression and trametinib sensitivity","authors":"Yanping Zhang , Shazhou Ye , Suying Wang , Qi Ding , Jing Jin , Ming Zhao","doi":"10.1016/j.prp.2025.156287","DOIUrl":"10.1016/j.prp.2025.156287","url":null,"abstract":"<div><div>Bladder cancer (BCa) is the most common malignancy of the urinary system. Despite advancements in novel targeted therapies and immunotherapy, the majority of patients remain incurable, and disease progression frequently occurs after treatment. Therefore, identifying new therapeutic strategies is crucial. Fatty acids are essential components of cell structure, playing roles in energy storage and serving as signaling molecules. In tumor tissues, due to abnormal blood vessel development, cancer cells primarily rely on de novo fatty acid synthesis to meet the demands of growth and proliferation. Stearoyl-CoA desaturase 1 (SCD1) is a key enzyme, widely recognized as a potential therapeutic target in various cancers. SCD1 promotes the synthesis of cell membranes by converting saturated fatty acids into monounsaturated fatty acids, thus supporting tumor cell growth. In this study, we conducted bioinformatics analysis using public datasets (including bulk RNA-seq and single-cell RNA-seq) and immunohistochemical examination of BCa tissues. Our findings reveal that SCD1 is specifically expressed in BCa cells and is associated with poor tumor grade and prognosis. Furthermore, drug sensitivity predictions and validations suggest that SCD1 enhances the sensitivity of BCa cells to trametinib. Therefore, SCD1 offers a promising new avenue for the early diagnosis, prognostic assessment, and optimization of personalized treatment strategies for BCa.</div></div>","PeriodicalId":19916,"journal":{"name":"Pathology, research and practice","volume":"277 ","pages":"Article 156287"},"PeriodicalIF":3.2,"publicationDate":"2025-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145615958","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Diabetic Kidney Disease (DKD) is a significant complication and leading cause of death in both type 1 and type 2 diabetes, as well as the primary cause of chronic kidney disease. Macrophage migration inhibitory factor (MIF) activates the NLRP3 inflammasome. Chicago sky blue 6B (CSB6B) is a MIF inhibitor with therapeutic potential in various inflammatory diseases, but its effect on DKD remains unexplored.
Materials and methods
HK-2 cell was used as the in vitro cell model. For the in vivo animal model, The db/db mice were randomly divided into three subgroups: the diabetic nephropathy model group, the low-dose CSB6B intervention group (2 mg/kg), and the high-dose CSB6B intervention group (8 mg/kg), with drug administration via intraperitoneal injection twice weekly for 12 weeks. CCK-8 assessed CSB6B toxicity, while qPCR measured MIF mRNA expression. Western blot, immunohistochemistry and ELISA detected protein expression level, and LDH release assessed membrane integrity. Histological analysis evaluated renal pathological changes.
Results
CSB6B significantly inhibited the secretion of inflammatory cytokines interleukin-1β (IL-1β) and TGF-β1 from high-glucose-stimulated HK-2 cells without affecting their viability. CSB6B effectively inhibited the expression and secretion of MIF in high-glucose-stimulated HK-2 cells, down-regulated the expression of NLRP3, suppressed the activation of NLRP3 inflammasomes, reduced the production of cell pyroptosis-related proteins, and significantly decreased collagen I and FN expression. CSB6B treatment significantly reduced the body weight, blood glucose, blood creatinine, urine ACR, and NGAL of db/db mice, and improved the pathological damage of diabetic nephropathy. CSB6B effectively reduced the expression level of MIF protein in diabetic nephropathy mice, down-regulated the expression of NLRP3, Caspase-1, GSDMD, IL-1β, Collagen I and FN in the renal cortex of diabetic nephropathy mice.
Conclusions
CSB6B mitigated DKD by inhibiting the NLRP3/Caspase-1/GSDMD pyroptosis signaling pathway, suppressed cell pyroptosis, reduced cytokine secretion, and decreasd extracellular matrix accumulation. CSB6B showed promise as a potential therapeutic for DKD.
{"title":"CSB6B attenuates renal inflammation and fibrosis by inhibiting the activation of NLRP3 inflammasome through the NLRP3/Caspase-1/GSDMD/IL-1β signaling pathway","authors":"Shuo Chen , Yonghong Zhu , Tong Chen , Yanyan Xu , Qiuling Fan","doi":"10.1016/j.prp.2025.156286","DOIUrl":"10.1016/j.prp.2025.156286","url":null,"abstract":"<div><h3>Context</h3><div>Diabetic Kidney Disease (DKD) is a significant complication and leading cause of death in both type 1 and type 2 diabetes, as well as the primary cause of chronic kidney disease. Macrophage migration inhibitory factor (MIF) activates the NLRP3 inflammasome. Chicago sky blue 6B (CSB6B) is a MIF inhibitor with therapeutic potential in various inflammatory diseases, but its effect on DKD remains unexplored.</div></div><div><h3>Materials and methods</h3><div>HK-2 cell was used as the in vitro cell model. For the in vivo animal model, The db/db mice were randomly divided into three subgroups: the diabetic nephropathy model group, the low-dose CSB6B intervention group (2 mg/kg), and the high-dose CSB6B intervention group (8 mg/kg), with drug administration via intraperitoneal injection twice weekly for 12 weeks. CCK-8 assessed CSB6B toxicity, while qPCR measured MIF mRNA expression. Western blot, immunohistochemistry and ELISA detected protein expression level, and LDH release assessed membrane integrity. Histological analysis evaluated renal pathological changes.</div></div><div><h3>Results</h3><div>CSB6B significantly inhibited the secretion of inflammatory cytokines interleukin-1β (IL-1β) and TGF-β1 from high-glucose-stimulated HK-2 cells without affecting their viability. CSB6B effectively inhibited the expression and secretion of MIF in high-glucose-stimulated HK-2 cells, down-regulated the expression of NLRP3, suppressed the activation of NLRP3 inflammasomes, reduced the production of cell pyroptosis-related proteins, and significantly decreased collagen I and FN expression. CSB6B treatment significantly reduced the body weight, blood glucose, blood creatinine, urine ACR, and NGAL of db/db mice, and improved the pathological damage of diabetic nephropathy. CSB6B effectively reduced the expression level of MIF protein in diabetic nephropathy mice, down-regulated the expression of NLRP3, Caspase-1, GSDMD, IL-1β, Collagen I and FN in the renal cortex of diabetic nephropathy mice.</div></div><div><h3>Conclusions</h3><div>CSB6B mitigated DKD by inhibiting the NLRP3/Caspase-1/GSDMD pyroptosis signaling pathway, suppressed cell pyroptosis, reduced cytokine secretion, and decreasd extracellular matrix accumulation. CSB6B showed promise as a potential therapeutic for DKD.</div></div>","PeriodicalId":19916,"journal":{"name":"Pathology, research and practice","volume":"277 ","pages":"Article 156286"},"PeriodicalIF":3.2,"publicationDate":"2025-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145518577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-07DOI: 10.1016/j.prp.2025.156284
Naglaa F. Khedr , Hend M. Selim , Gamal A. Abourayya
Chronic diseases like neurodegenerative disorders, musculoskeletal issues, metabolic diseases, cancer, liver and kidney disorders are increasingly linked to mitochondrial dysfunction. PINK1-Parkin-mediated mitophagy, a vital autophagic process, plays a central role in maintaining cellular homeostasis by selectively eliminating damaged mitochondria, which is crucial for preserving mitochondrial integrity and preventing reactive oxygen species accumulation. Activation of the PINK1-Parkin signaling pathway has emerged as a promising therapeutic strategy to restore mitochondrial function and attenuate disease progression. Recent studies have demonstrated that natural PINK1-Parkin activators offer significant therapeutic potential for treating a wide range of chronic diseases by modulating mitochondrial dynamics, alleviating cellular inflammation, and preventing mitochondrial damage. This review provides an in-depth analysis of the molecular mechanisms underlying PINK1-Parkin signaling, discusses the therapeutic benefits of natural activators, and presents them as a compelling strategy for addressing mitochondrial dysfunction and mitigating the progression of chronic diseases.
{"title":"Therapeutic potential of natural compounds in the management of chronic diseases: Targeting PINK1–Parkin pathway","authors":"Naglaa F. Khedr , Hend M. Selim , Gamal A. Abourayya","doi":"10.1016/j.prp.2025.156284","DOIUrl":"10.1016/j.prp.2025.156284","url":null,"abstract":"<div><div>Chronic diseases like neurodegenerative disorders, musculoskeletal issues, metabolic diseases, cancer, liver and kidney disorders are increasingly linked to mitochondrial dysfunction. PINK1-Parkin-mediated mitophagy, a vital autophagic process, plays a central role in maintaining cellular homeostasis by selectively eliminating damaged mitochondria, which is crucial for preserving mitochondrial integrity and preventing reactive oxygen species accumulation. Activation of the PINK1-Parkin signaling pathway has emerged as a promising therapeutic strategy to restore mitochondrial function and attenuate disease progression. Recent studies have demonstrated that natural PINK1-Parkin activators offer significant therapeutic potential for treating a wide range of chronic diseases by modulating mitochondrial dynamics, alleviating cellular inflammation, and preventing mitochondrial damage. This review provides an in-depth analysis of the molecular mechanisms underlying PINK1-Parkin signaling, discusses the therapeutic benefits of natural activators, and presents them as a compelling strategy for addressing mitochondrial dysfunction and mitigating the progression of chronic diseases.</div></div>","PeriodicalId":19916,"journal":{"name":"Pathology, research and practice","volume":"277 ","pages":"Article 156284"},"PeriodicalIF":3.2,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145506307","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-07DOI: 10.1016/j.prp.2025.156285
Muhammad Liaquat Raza , Mustafa Hussain Imam , Warisha Zehra , Insa Binte Anwar , Rukhsar Mehdi
Drug-resistant epilepsy (DRE) still poses one of the greatest therapeutic challenges, afflicting about one-third of all patients with epilepsy in the world. Despite spectacular advances in the fields of anti-seizure medications and neurostimulation techniques, treatment outcomes in DRE have reached plateau levels, signifying an urgent need for better mechanistic understanding and therapeutic strategies. New evidence increasingly elucidates mitochondrial dysfunction as a lens through which to understand seizure generation, pharmacoresistance, and disease progression. Mitochondria are regulators of ATP production, calcium buffering, and redox homeostasis; disruption of any such pathway will result in neuronal hyperexcitability, oxidative injury, and cell death. Moreover, mitochondrial DNA mutations and heteroplasmy threshold can correlate with seizure onset, seizure severity, and Response to treatment, thus being potential biomarkers for risk stratification. This narrative review surveys both preclinical and clinical evidence for mitochondrial dysfunction in epilepsy, examining oxidative stress pathways, mitophagy, and mitochondrial permeability transition pore opening as key mechanisms of neuronal vulnerability. We subsequently analyze various preclinical models of mitochondrial dysfunction, pointing out their respective strengths and weaknesses. Emerging therapeutic strategies, encompassing pharmacological agents, gene therapy, diet, and natural compounds, are then reviewed, which aim to resolve issues surrounding mitochondrial health on a molecular basis. By straddling the mechanistic and clinical narratives, this work foregrounds mitochondrial-centered approaches as promises for both the diagnostic and therapeutic arsenal in the management of DRE.
{"title":"Oxidative stress and neuronal alteration: Mitochondrial dysfunction as a key player in intractable epilepsy - a narrative review","authors":"Muhammad Liaquat Raza , Mustafa Hussain Imam , Warisha Zehra , Insa Binte Anwar , Rukhsar Mehdi","doi":"10.1016/j.prp.2025.156285","DOIUrl":"10.1016/j.prp.2025.156285","url":null,"abstract":"<div><div>Drug-resistant epilepsy (DRE) still poses one of the greatest therapeutic challenges, afflicting about one-third of all patients with epilepsy in the world. Despite spectacular advances in the fields of anti-seizure medications and neurostimulation techniques, treatment outcomes in DRE have reached plateau levels, signifying an urgent need for better mechanistic understanding and therapeutic strategies. New evidence increasingly elucidates mitochondrial dysfunction as a lens through which to understand seizure generation, pharmacoresistance, and disease progression. Mitochondria are regulators of ATP production, calcium buffering, and redox homeostasis; disruption of any such pathway will result in neuronal hyperexcitability, oxidative injury, and cell death. Moreover, mitochondrial DNA mutations and heteroplasmy threshold can correlate with seizure onset, seizure severity, and <strong>Response</strong> to treatment, thus being potential biomarkers for risk stratification. This narrative review surveys both preclinical and clinical evidence for mitochondrial dysfunction in epilepsy, examining oxidative stress pathways, mitophagy, and mitochondrial permeability transition pore opening as key mechanisms of neuronal vulnerability. We subsequently analyze various preclinical models of mitochondrial dysfunction, pointing out their respective strengths and weaknesses. Emerging therapeutic strategies, encompassing pharmacological agents, gene therapy, diet, and natural compounds, are then reviewed, which aim to resolve issues surrounding mitochondrial health on a molecular basis. By straddling the mechanistic and clinical narratives, this work foregrounds mitochondrial-centered approaches as promises for both the diagnostic and therapeutic arsenal in the management of DRE.</div></div>","PeriodicalId":19916,"journal":{"name":"Pathology, research and practice","volume":"277 ","pages":"Article 156285"},"PeriodicalIF":3.2,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145518578","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-05DOI: 10.1016/j.prp.2025.156278
Maccio Livia , Bragantini Emma , Piermattei Alessia , Santoro Angela , Zannoni Gian Franco
Uterine carcinosarcomas (UCS), also known as malignant mixed Müllerian tumors, are rare and aggressive neoplasms characterized by the coexistence of carcinomatous (epithelial) and sarcomatous (mesenchymal) components. PTEN (Phosphatase and Tensin Homolog) mutations, a hallmark of endometrioid-type carcinomas, play a significant role in the pathogenesis and histological presentation of a subset of uterine carcinosarcomas, contributing to their aggressive behavior and poor prognosis. This overview synthesizes the current understanding of PTEN mutations in carcinosarcomas and their implications for clinical management and therapy. A new approach based on morphological, immunohistochemical, and molecular characteristics is proposed.
子宫癌肉瘤（UCS），又称恶性混合性勒氏瘤，是一种罕见的侵袭性肿瘤，其特征是癌（上皮）和肉瘤（间充质）成分共存。PTEN （Phosphatase and Tensin Homolog）突变是子宫内膜样癌的一个标志，在子宫癌肉瘤的发病机制和组织学表现中起重要作用，导致其侵袭性行为和不良预后。本综述综合了目前对癌肉瘤中PTEN突变的理解及其对临床管理和治疗的意义。提出了一种基于形态学、免疫组织化学和分子特征的新方法。
{"title":"PTEN mutation status in uterine carcinosarcomas: A comprehensive overview and a new diagnostic approach","authors":"Maccio Livia , Bragantini Emma , Piermattei Alessia , Santoro Angela , Zannoni Gian Franco","doi":"10.1016/j.prp.2025.156278","DOIUrl":"10.1016/j.prp.2025.156278","url":null,"abstract":"<div><div>Uterine carcinosarcomas (UCS), also known as malignant mixed Müllerian tumors, are rare and aggressive neoplasms characterized by the coexistence of carcinomatous (epithelial) and sarcomatous (mesenchymal) components. PTEN (Phosphatase and Tensin Homolog) mutations, a hallmark of endometrioid-type carcinomas, play a significant role in the pathogenesis and histological presentation of a subset of uterine carcinosarcomas, contributing to their aggressive behavior and poor prognosis. This overview synthesizes the current understanding of PTEN mutations in carcinosarcomas and their implications for clinical management and therapy. A new approach based on morphological, immunohistochemical, and molecular characteristics is proposed.</div></div>","PeriodicalId":19916,"journal":{"name":"Pathology, research and practice","volume":"276 ","pages":"Article 156278"},"PeriodicalIF":3.2,"publicationDate":"2025-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145467116","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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