Pub Date : 2025-11-10DOI: 10.1016/j.yexcr.2025.114827
Qiang Xu , Ge Xuan , Chongying Zhu , Bingquan Zhu , Zihao An , Meiyuan Jin , Chao Tang
Preeclampsia (PE) is a multisystem disorder with increased maternal and perinatal mortality and morbidity while the treatment of PE remains largely unknown in clinic. Therefore, it is necessary to find more effective therapeutic methods of PE. We report herein that, Tianma Gouteng Decoction (TGD) generates protective effects against PE by activating Wnt 3a/β-catenin signaling and simultaneously suppressing ferroptosis in placental trophoblast cells. Particularly, Gastrodin (Gtd), the active component in TGD, plays a major role in protection against PE. Mechanistically, Gtd up-regulates the Wnt 3a/β-catenin signaling activity by inducing Wnt 3a mRNA expression, resulting in the increased expression of β-catenin-controlled target genes. On the other hand, Gtd-triggered Wnt activation obviously exerts negative effects on ferroptosis by promoting expression levels of the anti-ferroptosis proteins accompanied by the down-regulated reactive oxygen species (ROS) production and total iron content but the up-regulated L-Glutathione (GSH) levels. Consistently, Gtd-administration reveals apparent anti-hypertensive effects in a PE-like mouse model with diminished ferroptosis, whereas deactivation of β-catenin by administration with the specific antagonist ICG001 disrupts the protective effects derived from Gtd. Therefore, our results provide an innovative basis for the role of Gtd as a new therapy for PE.
{"title":"Gastrodin produces therapeutic effects against preeclampsia by activating Wnt 3a signaling and inhibiting ferroptosis","authors":"Qiang Xu , Ge Xuan , Chongying Zhu , Bingquan Zhu , Zihao An , Meiyuan Jin , Chao Tang","doi":"10.1016/j.yexcr.2025.114827","DOIUrl":"10.1016/j.yexcr.2025.114827","url":null,"abstract":"<div><div>Preeclampsia (PE) is a multisystem disorder with increased maternal and perinatal mortality and morbidity while the treatment of PE remains largely unknown in clinic. Therefore, it is necessary to find more effective therapeutic methods of PE. We report herein that, Tianma Gouteng Decoction (TGD) generates protective effects against PE by activating Wnt 3a/β-catenin signaling and simultaneously suppressing ferroptosis in placental trophoblast cells. Particularly, Gastrodin (Gtd), the active component in TGD, plays a major role in protection against PE. Mechanistically, Gtd up-regulates the Wnt 3a/β-catenin signaling activity by inducing <em>Wnt 3a</em> mRNA expression, resulting in the increased expression of β-catenin-controlled target genes. On the other hand, Gtd-triggered Wnt activation obviously exerts negative effects on ferroptosis by promoting expression levels of the anti-ferroptosis proteins accompanied by the down-regulated reactive oxygen species (ROS) production and total iron content but the up-regulated L-Glutathione (GSH) levels. Consistently, Gtd-administration reveals apparent anti-hypertensive effects in a PE-like mouse model with diminished ferroptosis, whereas deactivation of β-catenin by administration with the specific antagonist ICG001 disrupts the protective effects derived from Gtd. Therefore, our results provide an innovative basis for the role of Gtd as a new therapy for PE.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"454 1","pages":"Article 114827"},"PeriodicalIF":3.5,"publicationDate":"2025-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145502588","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 : 2025-11-06DOI: 10.1016/j.yexcr.2025.114816
Shuai Wu , Bin Li , Fulin Tao , Wenhao Song , Guoming Zhang , Rui Wang , Zhoubin Tian
Objective
Despite the high prevalence of osteoarthritis (OA), molecular mechanisms underlying cartilage destruction remain incompletely understood. This study investigated the expression pattern and functional role of muscle segment homeobox 2 (MSX2) in osteoarthritis pathogenesis.
Methods
Differential gene expression analysis was performed using three independent Gene Expression Omnibus (GEO) datasets (GSE55457, GSE12021, and GSE114007) to examine MSX2 expression in osteoarthritic versus normal cartilage. Expression patterns were validated in primary chondrocytes from human osteoarthritis patients and mouse osteoarthritis models using Western blot and quantitative reverse transcription polymerase chain reaction (RT-PCR). Functional studies employed both loss-of-function and gain-of-function approaches through intra-articular injection of adeno-associated virus expressing MSX2-specific short hairpin ribonucleic acid (shRNA) or MSX2 overexpression constructs. Cartilage morphology, proteoglycan content, chondrocyte apoptosis, matrix markers, and inflammatory responses were assessed through histological analysis, immunohistochemistry, and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) staining.
Results
MSX2 was consistently downregulated in osteoarthritic samples across all datasets, with validation confirming reduced MSX2 protein and messenger ribonucleic acid (mRNA) levels in both human patients and mouse models (p < 0.05). MSX2 knockdown significantly exacerbated cartilage degeneration, increased chondrocyte apoptosis, and disrupted matrix homeostasis by reducing anabolic markers type II collagen (COL2) and aggrecan (ACAN) while upregulating matrix metalloproteinase 13 (MMP13). Additionally, MSX2 knockdown enhanced inflammatory responses, evidenced by increased macrophage infiltration and tumor necrosis factor-alpha (TNFα) expression. Conversely, MSX2 overexpression demonstrated chondroprotective effects with improved cartilage morphology and reduced apoptosis.
Conclusion
MSX2 serves as a critical chondroprotective factor maintaining cartilage homeostasis. Its downregulation contributes to osteoarthritis pathogenesis through disruption of matrix homeostasis, promotion of chondrocyte apoptosis, and enhancement of inflammatory responses. These findings identify MSX2 as a potential therapeutic target for osteoarthritis treatment.
{"title":"MSX2 serves as a chondroprotective factor in osteoarthritis: insights from expression analysis and functional validation studies","authors":"Shuai Wu , Bin Li , Fulin Tao , Wenhao Song , Guoming Zhang , Rui Wang , Zhoubin Tian","doi":"10.1016/j.yexcr.2025.114816","DOIUrl":"10.1016/j.yexcr.2025.114816","url":null,"abstract":"<div><h3>Objective</h3><div>Despite the high prevalence of osteoarthritis (OA), molecular mechanisms underlying cartilage destruction remain incompletely understood. This study investigated the expression pattern and functional role of muscle segment homeobox 2 (MSX2) in osteoarthritis pathogenesis.</div></div><div><h3>Methods</h3><div>Differential gene expression analysis was performed using three independent Gene Expression Omnibus (GEO) datasets (GSE55457, GSE12021, and GSE114007) to examine MSX2 expression in osteoarthritic versus normal cartilage. Expression patterns were validated in primary chondrocytes from human osteoarthritis patients and mouse osteoarthritis models using Western blot and quantitative reverse transcription polymerase chain reaction (RT-PCR). Functional studies employed both loss-of-function and gain-of-function approaches through intra-articular injection of adeno-associated virus expressing MSX2-specific short hairpin ribonucleic acid (shRNA) or MSX2 overexpression constructs. Cartilage morphology, proteoglycan content, chondrocyte apoptosis, matrix markers, and inflammatory responses were assessed through histological analysis, immunohistochemistry, and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) staining.</div></div><div><h3>Results</h3><div>MSX2 was consistently downregulated in osteoarthritic samples across all datasets, with validation confirming reduced MSX2 protein and messenger ribonucleic acid (mRNA) levels in both human patients and mouse models (p < 0.05). MSX2 knockdown significantly exacerbated cartilage degeneration, increased chondrocyte apoptosis, and disrupted matrix homeostasis by reducing anabolic markers type II collagen (COL2) and aggrecan (ACAN) while upregulating matrix metalloproteinase 13 (MMP13). Additionally, MSX2 knockdown enhanced inflammatory responses, evidenced by increased macrophage infiltration and tumor necrosis factor-alpha (TNFα) expression. Conversely, MSX2 overexpression demonstrated chondroprotective effects with improved cartilage morphology and reduced apoptosis.</div></div><div><h3>Conclusion</h3><div>MSX2 serves as a critical chondroprotective factor maintaining cartilage homeostasis. Its downregulation contributes to osteoarthritis pathogenesis through disruption of matrix homeostasis, promotion of chondrocyte apoptosis, and enhancement of inflammatory responses. These findings identify MSX2 as a potential therapeutic target for osteoarthritis treatment.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"454 1","pages":"Article 114816"},"PeriodicalIF":3.5,"publicationDate":"2025-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145476701","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 : 2025-11-04DOI: 10.1016/j.yexcr.2025.114815
Jingyi Li , Xinping Zhu , Liang Feng , Xiaoqin Liu , YuQin Xiao , Yanting Zhang , Changlian Lu , Weiming Zhao
Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer, and chemotherapy remains a major component of the standard therapy. Although there is an initial response, the tumor will inevitably develop resistance to paclitaxel (PTX) treatment. Berberine (BBR) exhibits excellent anti-cancer activity in a variety of tumor cells and drug-resistant tumor cells. In the in vitro experiments of the study, the effect of BBR on PTX resistance was explored through proliferation assay, flow cytometry. The effects of combined treatment of BBR and PTX on DNA damage in TNBC cells were evaluated by colony formation assay, comet assay and γ-H2AX staining. Western blot and immunofluorescence revealed that HSP90 was highly expressed in the tolerant strain. BBR promotes the ubiquitination and degradation of HSP90 through the ubiquitin-protease system, further inhibits the expression of its client protein p-BRCA1 and improves the sensitivity of PTX treatment. Cell lines overexpressing hsp90 were constructed and it was found that Hsp90 could reverse the sensitivity of BBR to PTX. In addition, a model of transplanted tumors in nude mice was constructed to further verify the regulatory effect of BBR on PTX resistance in TNBC. Collectively, our findings demonstrate that BBR targets HSP90 to activate p-BRCA1-mediated DNA damage, thereby sensitizing non-BRCA-mutated TNBC to PTX. These results suggest HSP90 inhibition as a therapeutic strategy to circumvent chemotherapy resistance and a potential biomarker for predicting TNBC treatment response.
{"title":"Berberine overcomes paclitaxel resistance in triple-negative breast cancer by inhibiting HSP90-mediated BRCA1 phosphorylation","authors":"Jingyi Li , Xinping Zhu , Liang Feng , Xiaoqin Liu , YuQin Xiao , Yanting Zhang , Changlian Lu , Weiming Zhao","doi":"10.1016/j.yexcr.2025.114815","DOIUrl":"10.1016/j.yexcr.2025.114815","url":null,"abstract":"<div><div>Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer, and chemotherapy remains a major component of the standard therapy. Although there is an initial response, the tumor will inevitably develop resistance to paclitaxel (PTX) treatment. Berberine (BBR) exhibits excellent anti-cancer activity in a variety of tumor cells and drug-resistant tumor cells. In the in vitro experiments of the study, the effect of BBR on PTX resistance was explored through proliferation assay, flow cytometry. The effects of combined treatment of BBR and PTX on DNA damage in TNBC cells were evaluated by colony formation assay, comet assay and γ-H2AX staining. Western blot and immunofluorescence revealed that HSP90 was highly expressed in the tolerant strain. BBR promotes the ubiquitination and degradation of HSP90 through the ubiquitin-protease system, further inhibits the expression of its client protein p-BRCA1 and improves the sensitivity of PTX treatment. Cell lines overexpressing hsp90 were constructed and it was found that Hsp90 could reverse the sensitivity of BBR to PTX. In addition, a model of transplanted tumors in nude mice was constructed to further verify the regulatory effect of BBR on PTX resistance in TNBC. Collectively, our findings demonstrate that BBR targets HSP90 to activate p-BRCA1-mediated DNA damage, thereby sensitizing non-BRCA-mutated TNBC to PTX. These results suggest HSP90 inhibition as a therapeutic strategy to circumvent chemotherapy resistance and a potential biomarker for predicting TNBC treatment response.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"454 1","pages":"Article 114815"},"PeriodicalIF":3.5,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145458049","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 : 2025-11-03DOI: 10.1016/j.yexcr.2025.114814
Carmen Llorens-Cebrià , Daphne Bouwens , Max Van Der Velde , Mónica Duran , Mireia Salvadó-Pau , Irene Martínez-Díaz , Jordi Vilardell-Vilà , Anna Meseguer , Rafael Kramann , Roser Ferrer-Costa , Maria José Soler , Joan López-Hellín , Gerard Cantero-Recasens , Jitske Jansen , Conxita Jacobs-Cachá
Idiopathic nephrotic syndrome (INS) associated to focal segmental glomeruloesclerosis or minimal change disease is characterized by the presence of heavy levels of proteinuria. Filtrated proteins are normally actively reabsorbed in the proximal tubule by the megalin-cubilin-amnionless complex, located at the apical membrane of the proximal tubule epithelial cells. Megalin has a transmembrane domain but cubilin needs to interact with amnionless to reach the cell membrane in a complex known as CubAm. While megalin has a wide variety of ligands only a few proteins are exclusively transported by binding CubAm; one of those is Apolipoprotein A-I (ApoA-I). The reabsorption of Apolipoprotein A-I (ApoA-I) seems to be impaired in INS but this aspect of INS is difficult to study due to the low expression of CubAm in cultured proximal tubular cells as well as in other in vitro tubular modelling approaches. Here we show that RPTEC/TERT-1 cells cultured in monolayer barely express cubilin and amnionless proteins. In contrast, proximal tubular cells of induced Pluripotent Stem Cells (iPSCs)-derived kidney organoids showed robust expression of CubAm. In addition, protein overload induced an increase of the number of proximal tubular cells expressing cubilin that is reversed when the stimuli is removed. Finally, exogenously added ApoA-I targets cubilin suggesting that the CubAm complex is functional in our iPSCs-derived kidney organoids. Thus, kidney organoids provide a valuable system for modelling specific aspects of INS, including the impact of protein overload on tubular cells and the CubAm-mediated endocytosis of ApoA-I, which is thought to be impaired in these conditions.
{"title":"Modelling CubAm function and regulation in proximal tubular cells using iPSC-derived kidney organoids","authors":"Carmen Llorens-Cebrià , Daphne Bouwens , Max Van Der Velde , Mónica Duran , Mireia Salvadó-Pau , Irene Martínez-Díaz , Jordi Vilardell-Vilà , Anna Meseguer , Rafael Kramann , Roser Ferrer-Costa , Maria José Soler , Joan López-Hellín , Gerard Cantero-Recasens , Jitske Jansen , Conxita Jacobs-Cachá","doi":"10.1016/j.yexcr.2025.114814","DOIUrl":"10.1016/j.yexcr.2025.114814","url":null,"abstract":"<div><div>Idiopathic nephrotic syndrome (INS) associated to focal segmental glomeruloesclerosis or minimal change disease is characterized by the presence of heavy levels of proteinuria. Filtrated proteins are normally actively reabsorbed in the proximal tubule by the megalin-cubilin-amnionless complex, located at the apical membrane of the proximal tubule epithelial cells. Megalin has a transmembrane domain but cubilin needs to interact with amnionless to reach the cell membrane in a complex known as CubAm. While megalin has a wide variety of ligands only a few proteins are exclusively transported by binding CubAm; one of those is Apolipoprotein A-I (ApoA-I). The reabsorption of Apolipoprotein A-I (ApoA-I) seems to be impaired in INS but this aspect of INS is difficult to study due to the low expression of CubAm in cultured proximal tubular cells as well as in other <em>in vitro</em> tubular modelling approaches. Here we show that RPTEC/TERT-1 cells cultured in monolayer barely express cubilin and amnionless proteins. In contrast, proximal tubular cells of induced Pluripotent Stem Cells (iPSCs)-derived kidney organoids showed robust expression of CubAm. In addition, protein overload induced an increase of the number of proximal tubular cells expressing cubilin that is reversed when the stimuli is removed. Finally, exogenously added ApoA-I targets cubilin suggesting that the CubAm complex is functional in our iPSCs-derived kidney organoids. Thus, kidney organoids provide a valuable system for modelling specific aspects of INS, including the impact of protein overload on tubular cells and the CubAm-mediated endocytosis of ApoA-I, which is thought to be impaired in these conditions.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"453 2","pages":"Article 114814"},"PeriodicalIF":3.5,"publicationDate":"2025-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145450909","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 : 2025-10-30DOI: 10.1016/j.yexcr.2025.114811
Jiajia Wang , Shaokai Tang , Tingyi Yang , Fenghao Geng , Xiaoyou Tang , Rui Xia , Xin Zhang , Hao Bai , Hai Xiong , Daojiang Yu , Shuyu Zhang
Background
Melanoma is an aggressive malignancy with one of the highest mortality rates among skin cancers. Radiotherapy is a common treatment modality, but radioresistance remains a significant challenge. The stimulator of interferon genes (STING) pathway has been implicated in antitumor immunity and cancer treatment, yet its role in melanoma radiosensitivity is poorly understood.
Objective
This study aimed to investigate the role of STING in enhancing the radiosensitivity of cutaneous melanoma cells and to explore the underlying mechanisms involving reactive oxygen species (ROS) and the NLRP3 inflammasome.
Methods
Using TCGA database analysis, we examined the correlation between cGAS-STING pathway expression and melanoma patient survival. In vitro experiments were conducted on A375 and B16F10 melanoma cell lines, where STING was overexpressed or activated using the STING agonist cGAMP. Cell viability, apoptosis, ROS levels, and NLRP3/ASC complex activity were assessed following radiation treatment. In vivo studies involved tumor-bearing mice treated with cGAMP and radiation to evaluate tumor growth and survival.
Results
High expression of cGAS and STING was associated with improved survival in melanoma patients. STING overexpression or cGAMP treatment significantly reduced cell viability, increased ROS levels, and enhanced apoptosis in irradiated melanoma cells. Mechanistically, the activation of STING elevated NLRP3/ASC complex activity, and the NLRP3 inhibitor CY-09 reversed the radiosensitization effects. In vivo, cGAMP combined with radiation suppressed tumor growth and improved survival in melanoma-bearing mice.
Conclusion
STING enhances radiosensitivity of melanoma through ROS-NLRP3 axis, and combining STING agonists with radiotherapy may be a new strategy to overcome radioresistance.
{"title":"Activation of STING sensitizes melanoma cells to radiation through ROS-induced NLRP3","authors":"Jiajia Wang , Shaokai Tang , Tingyi Yang , Fenghao Geng , Xiaoyou Tang , Rui Xia , Xin Zhang , Hao Bai , Hai Xiong , Daojiang Yu , Shuyu Zhang","doi":"10.1016/j.yexcr.2025.114811","DOIUrl":"10.1016/j.yexcr.2025.114811","url":null,"abstract":"<div><h3>Background</h3><div>Melanoma is an aggressive malignancy with one of the highest mortality rates among skin cancers. Radiotherapy is a common treatment modality, but radioresistance remains a significant challenge. The stimulator of interferon genes (STING) pathway has been implicated in antitumor immunity and cancer treatment, yet its role in melanoma radiosensitivity is poorly understood.</div></div><div><h3>Objective</h3><div>This study aimed to investigate the role of STING in enhancing the radiosensitivity of cutaneous melanoma cells and to explore the underlying mechanisms involving reactive oxygen species (ROS) and the NLRP3 inflammasome.</div></div><div><h3>Methods</h3><div>Using TCGA database analysis, we examined the correlation between cGAS-STING pathway expression and melanoma patient survival. In vitro experiments were conducted on A375 and B16F10 melanoma cell lines, where STING was overexpressed or activated using the STING agonist cGAMP. Cell viability, apoptosis, ROS levels, and NLRP3/ASC complex activity were assessed following radiation treatment. In vivo studies involved tumor-bearing mice treated with cGAMP and radiation to evaluate tumor growth and survival.</div></div><div><h3>Results</h3><div>High expression of cGAS and STING was associated with improved survival in melanoma patients. STING overexpression or cGAMP treatment significantly reduced cell viability, increased ROS levels, and enhanced apoptosis in irradiated melanoma cells. Mechanistically, the activation of STING elevated NLRP3/ASC complex activity, and the NLRP3 inhibitor CY-09 reversed the radiosensitization effects. In vivo, cGAMP combined with radiation suppressed tumor growth and improved survival in melanoma-bearing mice.</div></div><div><h3>Conclusion</h3><div>STING enhances radiosensitivity of melanoma through ROS-NLRP3 axis, and combining STING agonists with radiotherapy may be a new strategy to overcome radioresistance.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"453 2","pages":"Article 114811"},"PeriodicalIF":3.5,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145414065","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}
Ferroptosis is an iron-dependent form of programmed cell death driven by the accumulation of lipid peroxides. KHSRP, an RNA-binding protein, is known to orchestrate diverse cellular processes, including cell differentiation, proliferation, and lipid metabolism. However, its potential role in modulating ferroptosis in cancer remains unclear. In this study, we found that elevated KHSRP expression was associated with poor prognosis in colorectal cancer (CRC) patients. Knockdown of KHSRP significantly elevated lipid peroxidation, increased malondialdehyde (MDA) accumulation, and reduced glutathione (GSH) levels, ultimately triggering ferroptosis in CRC cells. Mechanistically, we discovered that KHSRP interacts with the splicing factor hnRNPM, which directly binds to GPX4 mRNA. Critically, hnRNPM overexpression effectively rescued the decrease in GPX4 expression and the ferroptotic phenotype induced by KHSRP knockdown. These results suggest that the KHSRP-hnRNPM complex binds to GPX4 mRNA and acts as a key regulator of its post-transcriptional fate to sustain GPX4 expression. Overall, our results uncover a novel regulatory mechanism whereby high KHSRP expression protects CRC cells from ferroptosis. Targeting the KHSRP–hnRNPM-GPX4 axis to overcome ferroptosis resistance represents a promising therapeutic strategy for CRC.
{"title":"KHSRP protects colorectal cancer cells against ferroptosis by regulating GPX4 expression through interaction with hnRNPM","authors":"Xia Ding , Ningjing Zhang , Shuai Zhan , Wanjing Ding","doi":"10.1016/j.yexcr.2025.114812","DOIUrl":"10.1016/j.yexcr.2025.114812","url":null,"abstract":"<div><div>Ferroptosis is an iron-dependent form of programmed cell death driven by the accumulation of lipid peroxides. KHSRP, an RNA-binding protein, is known to orchestrate diverse cellular processes, including cell differentiation, proliferation, and lipid metabolism. However, its potential role in modulating ferroptosis in cancer remains unclear. In this study, we found that elevated KHSRP expression was associated with poor prognosis in colorectal cancer (CRC) patients. Knockdown of KHSRP significantly elevated lipid peroxidation, increased malondialdehyde (MDA) accumulation, and reduced glutathione (GSH) levels, ultimately triggering ferroptosis in CRC cells. Mechanistically, we discovered that KHSRP interacts with the splicing factor hnRNPM, which directly binds to GPX4 mRNA. Critically, hnRNPM overexpression effectively rescued the decrease in GPX4 expression and the ferroptotic phenotype induced by KHSRP knockdown. These results suggest that the KHSRP-hnRNPM complex binds to GPX4 mRNA and acts as a key regulator of its post-transcriptional fate to sustain GPX4 expression. Overall, our results uncover a novel regulatory mechanism whereby high KHSRP expression protects CRC cells from ferroptosis. Targeting the KHSRP–hnRNPM-GPX4 axis to overcome ferroptosis resistance represents a promising therapeutic strategy for CRC.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"453 2","pages":"Article 114812"},"PeriodicalIF":3.5,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145426534","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 : 2025-10-30DOI: 10.1016/j.yexcr.2025.114813
Yiyang Shu , Dandan Liu , Hai Xie , Chaoyang Zhang , Yanlong Bi , Jingfa Zhang
Purpose
To investigate the mechanisms underlying the microglia recruitment and its causal role in the breakdown of the outer blood-retinal barrier (oBRB) in diabetic retinopathy (DR).
Methods
The Sprague-Dawley rats were adopted to establish diabetic model by intraperitoneal injection of streptozotocin. Twelve weeks later, the retinal pigment epithelium (RPE)-choroid complexes and retinal paraffin sections were examined with immunofluorescence. RNA-sequencing was performed on glyoxal-treated ARPE-19 cells, followed by bioinformatic analysis to identify significant genes and pathways. Transwell assays were employed to establish the co-culture system and investigate the interactions between ARPE-19 and BV2 microglial cells. The results were further validated by the inhibitor or siRNAs targeting NF-κB, HO-1, and MCP-1.
Results
In 12-week diabetic rat retinas, microglia were observed to accumulate in the vicinity of the RPE cells, accompanied by the disruption of ZO-1. The expressions of ZO-1 and occludin remained largely unchanged in ARPE-19 cells when treated with glyoxal alone. However, when co-cultured with BV2 microglial cells, the expression levels of ZO-1 and occludin in glyoxal-treated ARPE-19 cells were significantly decreased, which were effectively prevented by siMCP-1. Mechanistically, RNA-sequencing analysis revealed that the activation of the NF-κB/HO-1/MCP-1 pathway in glyoxal-treated ARPE-19 cells significantly contributed to the recruitment of microglia. The above effects were reversed by BAY 11–7082, siHO-1 or siMCP-1.
Conclusion
Under diabetic conditions, microglia are recruited by RPE cells via the NF-κB/HO-1/MCP-1 pathway, which subsequently results in the oBRB breakdown. This study provides a novel mechanistic insight for the interaction between microglia and RPE cells, and implies a potential therapeutic strategy for the treatment of DR.
{"title":"The RPE-derived NF-κB/HO-1/MCP-1 pathway mediated microglia recruitment involved in the outer blood-retinal barrier breakdown in experimental diabetic retinopathy","authors":"Yiyang Shu , Dandan Liu , Hai Xie , Chaoyang Zhang , Yanlong Bi , Jingfa Zhang","doi":"10.1016/j.yexcr.2025.114813","DOIUrl":"10.1016/j.yexcr.2025.114813","url":null,"abstract":"<div><h3>Purpose</h3><div>To investigate the mechanisms underlying the microglia recruitment and its causal role in the breakdown of the outer blood-retinal barrier (oBRB) in diabetic retinopathy (DR).</div></div><div><h3>Methods</h3><div>The Sprague-Dawley rats were adopted to establish diabetic model by intraperitoneal injection of streptozotocin. Twelve weeks later, the retinal pigment epithelium (RPE)-choroid complexes and retinal paraffin sections were examined with immunofluorescence. RNA-sequencing was performed on glyoxal-treated ARPE-19 cells, followed by bioinformatic analysis to identify significant genes and pathways. Transwell assays were employed to establish the co-culture system and investigate the interactions between ARPE-19 and BV2 microglial cells. The results were further validated by the inhibitor or siRNAs targeting NF-κB, HO-1, and MCP-1.</div></div><div><h3>Results</h3><div>In 12-week diabetic rat retinas, microglia were observed to accumulate in the vicinity of the RPE cells, accompanied by the disruption of ZO-1. The expressions of ZO-1 and occludin remained largely unchanged in ARPE-19 cells when treated with glyoxal alone. However, when co-cultured with BV2 microglial cells, the expression levels of ZO-1 and occludin in glyoxal-treated ARPE-19 cells were significantly decreased, which were effectively prevented by siMCP-1. Mechanistically, RNA-sequencing analysis revealed that the activation of the NF-κB/HO-1/MCP-1 pathway in glyoxal-treated ARPE-19 cells significantly contributed to the recruitment of microglia. The above effects were reversed by BAY 11–7082, siHO-1 or siMCP-1.</div></div><div><h3>Conclusion</h3><div>Under diabetic conditions, microglia are recruited by RPE cells via the NF-κB/HO-1/MCP-1 pathway, which subsequently results in the oBRB breakdown. This study provides a novel mechanistic insight for the interaction between microglia and RPE cells, and implies a potential therapeutic strategy for the treatment of DR.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"454 1","pages":"Article 114813"},"PeriodicalIF":3.5,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145426546","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 : 2025-10-26DOI: 10.1016/j.yexcr.2025.114801
Han Zheng , Xiao Zhang , Mei Li , Yu Wang , Shufen Guo , Ming Jiang , Rui Xu , Yulin Huang , Zhengliang Ma
Obesity markedly exacerbates nociceptive sensitivity and substantially compromises the quality of life of affected people. Astrocytes orchestrate metabolic regulation and homeostatic maintenance in the central nervous system. Notably, fatty acid binding protein 7 (FABP7) is highly expressed in astrocytes that governs intracellular fatty acid uptake and transport. While systemic hyperlipidemia is pathognomonic of obesity, the mechanistic contribution of FABP7 in astrocytes to obesity-associated pain pathophysiology remains poorly characterized. The present study established a model of high-fat diet (HFD)-induced obesity combined with a standardized hind paw surgical incision paradigm, aiming to unveil the role of astrocytic FABP7 in HFD-induced chronic pain. Furthermore, an in vitro high-fat environment was induced by palmitic acid (PA),aiming to investigate the molecular mechanisms by which primary astrocytes were activated into the A1 phenotype to mediate neuroinflammation. FABP7 was overexpressed in the spinal dorsal of HFD mice. The activation of A1-type astrocytes and neuroinflammation cascades involving elevated iNOS eventually resulted in mechanical allodynia in HFD mice. Pharmacological inhibition of FABP7 via an intraperitoneal administration of SBFI-26 (20 mg/kg) significantly attenuated the paw withdrawal mechanical threshold and inhibited the A1-type astrocytes activation. PA-induced high-fat conditions promoted lipid droplet accumulation and upregulated FABP7 in astrocytes. Pharmacological inhibition of FABP7 using SBFI-26 (100 μmol/L) significantly suppressed the neurotoxic C3-positive A1 astrocyte phenotype, reduced intracellular lipid droplet accumulation, thereby inhibiting the activation of A1-type astrocytes and alleviating neuroinflammation. Overall, FABP7-mediated astrocytic reprogramming was a critical nexus bridging obesity with chronic pain. A1-astrocyte polarization initiated neuroinflammatory amplification, forming a feedforward loop perpetuating central sensitization. Our findings are expected to offer a viable target for metabolic pain management.
{"title":"The role and mechanism of fatty acid-binding protein 7 in spinal reactive astrocytes in prolonged postoperative pain induced by high-fat diet","authors":"Han Zheng , Xiao Zhang , Mei Li , Yu Wang , Shufen Guo , Ming Jiang , Rui Xu , Yulin Huang , Zhengliang Ma","doi":"10.1016/j.yexcr.2025.114801","DOIUrl":"10.1016/j.yexcr.2025.114801","url":null,"abstract":"<div><div>Obesity markedly exacerbates nociceptive sensitivity and substantially compromises the quality of life of affected people. Astrocytes orchestrate metabolic regulation and homeostatic maintenance in the central nervous system. Notably, fatty acid binding protein 7 (FABP7) is highly expressed in astrocytes that governs intracellular fatty acid uptake and transport. While systemic hyperlipidemia is pathognomonic of obesity, the mechanistic contribution of FABP7 in astrocytes to obesity-associated pain pathophysiology remains poorly characterized. The present study established a model of high-fat diet (HFD)-induced obesity combined with a standardized hind paw surgical incision paradigm, aiming to unveil the role of astrocytic FABP7 in HFD-induced chronic pain. Furthermore, an <em>in vitro</em> high-fat environment was induced by palmitic acid (PA),aiming to investigate the molecular mechanisms by which primary astrocytes were activated into the A1 phenotype to mediate neuroinflammation. FABP7 was overexpressed in the spinal dorsal of HFD mice. The activation of A1-type astrocytes and neuroinflammation cascades involving elevated iNOS eventually resulted in mechanical allodynia in HFD mice. Pharmacological inhibition of FABP7 <em>via</em> an intraperitoneal administration of SBFI-26 (20 mg/kg) significantly attenuated the paw withdrawal mechanical threshold and inhibited the A1-type astrocytes activation. PA-induced high-fat conditions promoted lipid droplet accumulation and upregulated FABP7 in astrocytes. Pharmacological inhibition of FABP7 using SBFI-26 (100 μmol/L) significantly suppressed the neurotoxic C3-positive A1 astrocyte phenotype, reduced intracellular lipid droplet accumulation, thereby inhibiting the activation of A1-type astrocytes and alleviating neuroinflammation. Overall, FABP7-mediated astrocytic reprogramming was a critical nexus bridging obesity with chronic pain. A1-astrocyte polarization initiated neuroinflammatory amplification, forming a feedforward loop perpetuating central sensitization. Our findings are expected to offer a viable target for metabolic pain management.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"453 2","pages":"Article 114801"},"PeriodicalIF":3.5,"publicationDate":"2025-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145388196","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}
Circadian rhythms regulate key biological processes, including cell proliferation and metabolism, and their disruption is implicated in colorectal cancer (CRC). mTOR signaling interacts bidirectionally with the circadian clock, yet how mTOR inhibition modulates clock gene dynamics and cellular behavior in intestinal models remains unclear. This study aimed to investigate the effects of everolimus, an mTOR inhibitor, on circadian gene expression, cell viability, apoptosis, and cell cycle progression in synchronized Caco-2 cells, with consideration of cell confluency and Circadian Time (CT).
Methods
Caco-2 cells were synchronized using serum shock at 20 % and 70 % confluency. Time-series samples were collected across multiple CTs (CT6–CT60). Gene expression (BMAL1, PER2, mTOR) was assessed by qRT-PCR using ACTB and RPLP0 as reference genes. Rhythmicity was evaluated via Cosinor analysis. Cell viability, apoptosis, and cell cycle dynamics were analyzed using the Muse™ Cell Analyzer following everolimus treatment (1–50 μM).
Results
RPLP0 proved to be a more stable reference gene than ACTB. BMAL1 exhibited stronger rhythmic expression than PER2, particularly at 20 % confluency. Everolimus (50 μM) significantly reduced cell viability in a time-dependent manner, with the greatest effect at CT6 and CT18 (p < 0.0001). Apoptosis was markedly increased at CT6 (+38.5 %) and moderate at CT18, indicating circadian modulation of drug sensitivity. Serum shock alone shifted cell cycle distribution, decreasing G0/G1 and increasing G2/M phase populations (p < 0.01). Everolimus altered BMAL1 and PER2 expression rhythms and significantly reduced mTOR expression at CT30, where baseline mTOR levels were highest. Cosinor analysis confirmed rhythmicity in BMAL1/RPLP0 and mTOR/RPLP0 profiles under low confluency.
Conclusion
Our findings demonstrate that everolimus influences circadian gene expression and exerts time-dependent antiproliferative and pro-apoptotic effects in Caco-2 cells. These results support the potential of circadian timing as a strategy to enhance mTOR-targeted therapies in CRC.
{"title":"Effects of everolimus on circadian gene expression and cell fate in synchronized Caco-2 cells","authors":"Dilek Ozturk Civelek , Beyza Goncu , Sadullah Goncu , Alper Okyar","doi":"10.1016/j.yexcr.2025.114810","DOIUrl":"10.1016/j.yexcr.2025.114810","url":null,"abstract":"<div><h3>Objectives</h3><div>Circadian rhythms regulate key biological processes, including cell proliferation and metabolism, and their disruption is implicated in colorectal cancer (CRC). mTOR signaling interacts bidirectionally with the circadian clock, yet how mTOR inhibition modulates clock gene dynamics and cellular behavior in intestinal models remains unclear. This study aimed to investigate the effects of everolimus, an mTOR inhibitor, on circadian gene expression, cell viability, apoptosis, and cell cycle progression in synchronized Caco-2 cells, with consideration of cell confluency and Circadian Time (CT).</div></div><div><h3>Methods</h3><div>Caco-2 cells were synchronized using serum shock at 20 % and 70 % confluency. Time-series samples were collected across multiple CTs (CT6–CT60). Gene expression (<em>BMAL1, PER2, mTOR</em>) was assessed by qRT-PCR using <em>ACTB</em> and <em>RPLP0</em> as reference genes. Rhythmicity was evaluated via Cosinor analysis. Cell viability, apoptosis, and cell cycle dynamics were analyzed using the Muse™ Cell Analyzer following everolimus treatment (1–50 μM).</div></div><div><h3>Results</h3><div><em>RPLP0</em> proved to be a more stable reference gene than <em>ACTB</em>. <em>BMAL1</em> exhibited stronger rhythmic expression than <em>PER2</em>, particularly at 20 % confluency. Everolimus (50 μM) significantly reduced cell viability in a time-dependent manner, with the greatest effect at CT6 and CT18 (p < 0.0001). Apoptosis was markedly increased at CT6 (+38.5 %) and moderate at CT18, indicating circadian modulation of drug sensitivity. Serum shock alone shifted cell cycle distribution, decreasing G0/G1 and increasing G2/M phase populations (p < 0.01). Everolimus altered <em>BMAL1</em> and <em>PER2</em> expression rhythms and significantly reduced mTOR expression at CT30, where baseline mTOR levels were highest. Cosinor analysis confirmed rhythmicity in <em>BMAL1/RPLP0</em> and <em>mTOR/RPLP0</em> profiles under low confluency.</div></div><div><h3>Conclusion</h3><div>Our findings demonstrate that everolimus influences circadian gene expression and exerts time-dependent antiproliferative and pro-apoptotic effects in Caco-2 cells. These results support the potential of circadian timing as a strategy to enhance mTOR-targeted therapies in CRC.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"453 2","pages":"Article 114810"},"PeriodicalIF":3.5,"publicationDate":"2025-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145414066","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 : 2025-10-24DOI: 10.1016/j.yexcr.2025.114799
Liu Yang , Yaodong Fan , Yiyin Wang , Chuntao Yan , Zheng Guan , Yuan Li , Xiaosan Su , Xiaowei Huang
Lung adenocarcinoma (LUAD), the predominant non-small cell lung cancer subtype, exhibits high mortality due to metastasis and therapeutic resistance. While circular RNAs (circRNAs) are implicated in oncogenesis, their functional mechanisms and upstream regulation in LUAD remain incompletely characterized. This study identifies circDNAJC16 (hsa_circ_0000018) as significantly downregulated in advanced-stage LUAD (Stage III-IV vs. I-II, p = 0.001), where its low expression independently predicts poor survival (HR = 1.93, p = 0.043). Functional characterization demonstrates that circDNAJC16 overexpression suppresses in vivo tumor growth (volume reduction: 26.56 %, p < 0.001) through cytoplasmic sequestration of oncogenic miR-93-5p, thereby activating CDKN1A/p21 to induce G0/G1 cell cycle arrest and inhibit proliferation, while concurrently suppressing metastasis via epithelial-mesenchymal transition (EMT) regulation. Crucially, the RNA-binding protein eIF4A3 binds upstream flanking introns of the host DNAJC16 pre-mRNA, driving selective nuclear retention of circDNAJC16 and redirecting linear DNAJC16 mRNA to the cytoplasm – a bifurcation mechanism essential for tumor suppression. These findings identify circDNAJC16 downregulation as a negative prognostic indicator in LUAD and reveal its dual tumor-suppressive roles: cytoplasmic sequestration of miR-93-5p activating CDKN1A-mediated cell cycle arrest, coupled with eIF4A3-governed nuclear retention controlling functional subcellular localization. Significantly, this work is the first to demonstrate eIF4A3-mediated circRNA compartmentalization, establishing circDNAJC16 as a novel prognostic biomarker and therapeutic target for LUAD.
{"title":"EIF4A3-mediated localization of circDNAJC16 sequesters miR-93-5p to suppress lung adenocarcinoma progression via CDKN1A-regulated cell cycle and EMT","authors":"Liu Yang , Yaodong Fan , Yiyin Wang , Chuntao Yan , Zheng Guan , Yuan Li , Xiaosan Su , Xiaowei Huang","doi":"10.1016/j.yexcr.2025.114799","DOIUrl":"10.1016/j.yexcr.2025.114799","url":null,"abstract":"<div><div>Lung adenocarcinoma (LUAD), the predominant non-small cell lung cancer subtype, exhibits high mortality due to metastasis and therapeutic resistance. While circular RNAs (circRNAs) are implicated in oncogenesis, their functional mechanisms and upstream regulation in LUAD remain incompletely characterized. This study identifies <strong>circDNAJC16 (hsa_circ_0000018)</strong> as significantly downregulated in advanced-stage LUAD (Stage III-IV vs. I-II, <em>p</em> = 0.001), where its low expression independently predicts poor survival (HR = 1.93, <em>p</em> = 0.043). Functional characterization demonstrates that circDNAJC16 overexpression suppresses <em>in vivo</em> tumor growth (volume reduction: 26.56 %, <em>p</em> < 0.001) through cytoplasmic sequestration of <strong>oncogenic miR-93-5p</strong>, thereby activating <strong>CDKN1A/p21</strong> to induce G0/G1 cell cycle arrest and inhibit proliferation, while concurrently suppressing metastasis via epithelial-mesenchymal transition (EMT) regulation. Crucially, the RNA-binding protein <strong>eIF4A3</strong> binds upstream flanking introns of the host DNAJC16 pre-mRNA, driving selective nuclear retention of circDNAJC16 and redirecting linear DNAJC16 mRNA to the cytoplasm – a bifurcation mechanism essential for tumor suppression. These findings identify circDNAJC16 downregulation as a negative prognostic indicator in LUAD and reveal its dual tumor-suppressive roles: cytoplasmic sequestration of miR-93-5p activating CDKN1A-mediated cell cycle arrest, coupled with eIF4A3-governed nuclear retention controlling functional subcellular localization. Significantly, this work is the first to demonstrate eIF4A3-mediated circRNA compartmentalization, establishing circDNAJC16 as a novel prognostic biomarker and therapeutic target for LUAD.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"453 2","pages":"Article 114799"},"PeriodicalIF":3.5,"publicationDate":"2025-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145370111","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}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号