Biochimica et biophysica acta. Molecular cell research最新文献

{"title":"RBM22-depletion delays progression through all steps of cell cycle and increases ploidy in myeloid cells","authors":"Eloïse Le Hir-Reynaud ,&nbsp;Benoît Soubise ,&nbsp;Abrahan Molina Mendoza ,&nbsp;Cassandra Konan ,&nbsp;Séverine Commet ,&nbsp;Nadia Gueganic ,&nbsp;Corinne Tous ,&nbsp;Laurent Corcos ,&nbsp;Nathalie Douet-Guilbert ,&nbsp;Marie-Bérengère Troadec","doi":"10.1016/j.bbamcr.2025.119965","DOIUrl":"10.1016/j.bbamcr.2025.119965","url":null,"abstract":"<div><div>RNA-Binding Motif 22 (<em>RBM22</em>) is a splicing factor and a transcription regulator that plays important roles in cancer. Our goal was to document further the implication of RBM22 in cell cycle progression. Using normal human haematopoietic stem and progenitor cells and myeloid cell lines (MDS-L, HL-60), we demonstrated that <em>RBM22</em> depletion reduces proliferation by delaying the progression of the G1-phase, S-phase and G2/M phase. <em>RBM22</em> depletion alters mitosis, generating endomitosis and alters megakaryocyte differentiation. Altogether, we propose, for the first time, <em>RBM22</em> as an essential actor of the cell cycle regulation in human haematopoietic stem and progenitor cells and myeloid cells. We demonstrated that <em>RBM22</em> alteration is partially responsible for the phenotype of cytopenia of myeloid cell lineages observed in myelodysplastic syndromes (MDS) with a partial deletion of chromosome 5 (MDS with del(5q)) where one allele of <em>RBM22</em> is lost. We hypothesise that the impact of <em>RBM22</em> on cell cycle progression could explain some phenotypic features of other cancers.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 6","pages":"Article 119965"},"PeriodicalIF":4.6,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143883010","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Non-canonical Wnt co-receptors ROR1/ROR2 are differentially regulated by hypoxia in colon cancer cells","authors":"Eduardo Alvarado-Ortiz ,&nbsp;María Cristina Castañeda-Patlán ,&nbsp;Angela Patricia Moreno-Londoño ,&nbsp;José Manuel Tinajero-Rodríguez ,&nbsp;Paola Briseño-Díaz ,&nbsp;Miguel Angel Sarabia-Sánchez ,&nbsp;Miguel Vargas ,&nbsp;Elizabeth Ortiz-Sánchez ,&nbsp;Martha Robles-Flores","doi":"10.1016/j.bbamcr.2025.119968","DOIUrl":"10.1016/j.bbamcr.2025.119968","url":null,"abstract":"<div><div>ROR1 and ROR2 co-receptors are transducers of non-canonical Wnt responses that promote an aggressive phenotype in several cancer types, including colon cancer. It has been demonstrated that hypoxia promotes tumor progression through the action of Hypoxia Inducible Factors (HIFs). An in silico analysis revealed that ROR2 is overexpressed in the advanced clinical stages of colon cancer. In line with this, ROR1 and ROR2 were found to be only expressed in malignant colon cells compared to non-malignant ones. The blockade of either ROR1 or ROR2 impaired colon cancer cells' colony formation abilities and the migration capacity of them. Additionally, the silencing of the ROR2 co-receptor blocked the metastatic ability of colon cancer cells in a xenografted mice model. We found that while silencing HIF-1α did not significantly reduce ROR1 or ROR2 expression, inhibiting HIF-2α and HIF-3α expression greatly decreased the protein levels of both co-receptors in colon cancer cells. The HIF-1α subunit expression is induced in acute hypoxia, whereas HIF-2α and HIF-3α show higher activity in chronic hypoxia, which may be functionally relevant since hypoxia induced a decrease in the constitutive active β-catenin transcriptional activity in SW480 cells. While both ROR1 and ROR2 stimulate proliferation and migration under normoxic conditions, the exposure of cells to hypoxia increased the expression of ROR1 or ROR2, depending on the Wnt cellular context, Thus, our results indicate that hypoxia partially represses β-catenin transcriptional activity and activates non-canonical Wnt signaling by regulating ROR1/ROR2 expression to induce an aggressive migrating and metastatic phenotype in colon cancer cells.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 5","pages":"Article 119968"},"PeriodicalIF":4.6,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143863862","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"OrgaMeas: A pipeline that integrates all the processes of organelle image analysis","authors":"Taiki Baba,&nbsp;Akimi Inoue,&nbsp;Susumu Tanimura,&nbsp;Kohsuke Takeda","doi":"10.1016/j.bbamcr.2025.119964","DOIUrl":"10.1016/j.bbamcr.2025.119964","url":null,"abstract":"<div><div>Although image analysis has emerged as a key technology in the study of organelle dynamics, the commonly used image-processing methods, such as threshold-based segmentation and manual setting of regions of interests (ROIs), are error-prone and laborious. Here, we present a highly accurate high-throughput image analysis pipeline called OrgaMeas for measuring the morphology and dynamics of organelles. This pipeline mainly consists of two deep learning-based tools: OrgaSegNet and DIC2Cells. OrgaSegNet quantifies many aspects of different organelles by precisely segmenting them. To further process the segmented data at a single-cell level, DIC2Cells automates ROI settings through accurate segmentation of individual cells in differential interference contrast (DIC) images. This pipeline was designed to be low cost and require less coding, to provide an easy-to-use platform. Thus, we believe that OrgaMeas has potential to be readily applied to basic biomedical research, and hopefully to other practical uses such as drug discovery.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 5","pages":"Article 119964"},"PeriodicalIF":4.6,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143869501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Intracellular and exosomal localization of the negative checkpoint regulator VISTA in immune cells","authors":"Lena Dübbel ,&nbsp;Anna Göken-Riebisch ,&nbsp;Karl-Wilhelm Koch","doi":"10.1016/j.bbamcr.2025.119966","DOIUrl":"10.1016/j.bbamcr.2025.119966","url":null,"abstract":"<div><div>Strategies in immunotherapy often target the immunosuppressive environment of tumor cells. One route of therapeutic interference could involve negative checkpoint regulators of which <u>V</u>-domain <u>i</u>mmunoglobulin (Ig)-containing <u>s</u>uppressor of <u>T</u>-cell <u>a</u>ctivation (VISTA) has raised more interest recently. The protein is expressed on the surface of tumor cells, T-lymphocytes, and antigen-presenting cells (APCs), but its intracellular expression pattern has not been investigated yet. We examined the intracellular distribution of VISTA and its possible role in translocation processes by immunofluorescence and Western blots.</div><div>We analyzed the expression and localization of VISTA in murine bone marrow-derived macrophages (BMDMs), human monocyte-derived macrophages, and human T lymphocytes (Jurkat). We obtained different cell fractions and organelles of various cell types and analyzed for the presence of VISTA. Monitoring a VISTA-GFP fusion construct in transfected cell lines HL-60 and THP-1 confirmed VISTA localization in these cell lines. All used cell lines showed the colocalization of VISTA and several vesicle markers together with VISTA staining along microtubule fibers. Additionally, we found VISTA in secreted exosomes and have the first hints for nucleic expression in all tested cell lines.</div><div>Therefore, the storage of VISTA in vesicles and its potential presence in nuclei resembles two other well-described checkpoint regulators, CTLA-4 and PD-L1, respectively.</div><div>We conclude that VISTA storage in vesicles enables a fast response to immunogenic stimuli, which needs to be considered for inhibitory experiments. The localization of VISTA in exosomes suggests a signaling function to facilitate cell-cell communication. Furthermore, the VISTA expression in the nucleus proposes a transcriptional role.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 5","pages":"Article 119966"},"PeriodicalIF":4.6,"publicationDate":"2025-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143859696","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Baicalein disrupts TGF-β-induced EMT in pancreatic cancer by FTO-dependent m6A demethylation of ZEB1","authors":"Lian Zhao,&nbsp;Gong Chen,&nbsp;Dan Li,&nbsp;Kangtao Wang,&nbsp;Michael Schaefer,&nbsp;Ingrid Herr ,&nbsp;Bin Yan","doi":"10.1016/j.bbamcr.2025.119969","DOIUrl":"10.1016/j.bbamcr.2025.119969","url":null,"abstract":"<div><div>Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy associated with poor prognosis. Baicalein, a flavonoid extracted from the roots of Scutellaria baicalensis, traditionally used in Chinese medicine, has demonstrated potential in inhibiting cancer development and progression. However, its mechanism of action remains poorly understood, particularly regarding epigenetic gene regulation through m6A RNA methylation. In this study, three human PDAC cell lines and one nonmalignant cell line were employed. The effects of baicalein were examined using multiple assays, including RT-qPCR, MeRIP-qPCR, Western blotting, spheroid formation, RNA stability, and MTT, to evaluate cellular functions and m6A regulation. Baicalein significantly reduced cell viability, migration, invasion, and colony formation. It also downregulated FTO, an enzyme critical for m6A RNA demethylation. Knockdown of FTO replicated the effects of baicalein, underscoring its oncogenic role in PDAC. Bioinformatic analysis identified ZEB1—a key transcription factor in epithelial-to-mesenchymal transition—as an m6A-modified target regulated by FTO. Both baicalein treatment and FTO knockdown enhanced m6A modification and decreased <em>ZEB1</em> mRNA stability, thereby suppressing stemness-related features. Rescue experiments further confirmed that baicalein disrupts the TGF-β/FTO/ZEB1 signaling axis, highlighting its therapeutic potential in PDAC. This study offers fundamental insights for the development of novel therapeutic strategies targeting PDAC.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 5","pages":"Article 119969"},"PeriodicalIF":4.6,"publicationDate":"2025-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143863867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Role and regulatory mechanism of DLX5 in rhabdomyosarcoma tumorigenesis","authors":"Yanxue Zhao ,&nbsp;Xinpei Liu ,&nbsp;Zining Wu ,&nbsp;Guotao Ma ,&nbsp;Quanli Gao ,&nbsp;Jun Zheng ,&nbsp;Chaoji Zhang","doi":"10.1016/j.bbamcr.2025.119959","DOIUrl":"10.1016/j.bbamcr.2025.119959","url":null,"abstract":"<div><div>Rhabdomyosarcoma (RMS), a common malignant tumor in children, presents numerous challenges in clinical treatment. This study investigated the specific functions and regulatory mechanisms of distal-less homeobox 5 (<em>DLX5</em>) in RMS. Data from TCGA, GEO and GEPIA databases were downloaded and analyzed. The effect of <em>DLX5</em> and <em>PAX3-FOXO1</em> on RMS cells was examined through cellular experiments. Binding activity between DLX5 and H3K9me2 was assessed using pull-down and chromatin immunoprecipitation-qPCR assays. Additionally, RMS model mice were constructed via xenotransplantation to validate the in vivo effects of <em>DLX5</em> on RMS. The results revealed that DLX5 was upregulated in RMS tissues and increased in various RMS cell lines, particularly in alveolar RMS cell lines. <em>DLX5</em> knockdown inhibited malignant biological behaviors. Besides, DLX5 expression was associated with myogenic differentiation of RMS cells. While the overexpression or knockdown of <em>DLX5</em> did not affect <em>PAX-FOXO1</em> expression. <em>PAX3-FOXO1</em> knockdown reduced DLX5 expression, indicating that <em>DLX5</em> act as a downstream effector of <em>PAX3-FOXO1</em>. Mechanistically, <em>PAX3-FOXO1</em> regulated DLX5 expression through <em>KDM4B/H3K9me2</em> axis. In vitro experiments further demonstrated that knockout of <em>DLX5</em> or <em>KDM4B</em> inhibited tumor growth. In conclusion, DLX5 expression was increased in <em>PAX3-FOXO1</em>–driven RMS, and its knockdown inhibited malignant biological behaviors of RMS cells. Moreover, the aberrant expression of DLX5 in <em>PAX3-FOXO1</em>–driven RMS was regulated by <em>KDM4B/H3K9me2</em> axis. These findings provided potential therapeutic targets for RMS treatment.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 5","pages":"Article 119959"},"PeriodicalIF":4.6,"publicationDate":"2025-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143854694","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"POLR1F promotes proliferation and stemness of anaplastic thyroid cancer by activating F2R/p38 MAPK signaling","authors":"Yingying Gong ,&nbsp;Shanshan Wang ,&nbsp;Ziwen Fang ,&nbsp;Xiaoping Hu ,&nbsp;Ying Li ,&nbsp;Yulu Che ,&nbsp;Zhuo Tan ,&nbsp;Baochun Su ,&nbsp;Minghua Ge ,&nbsp;Zongfu Pan","doi":"10.1016/j.bbamcr.2025.119963","DOIUrl":"10.1016/j.bbamcr.2025.119963","url":null,"abstract":"<div><div>Anaplastic thyroid cancer (ATC) is one of the most aggressive cancers characterized by a rapid growth rate. Dysregulation of RNA polymerase (Pol) is critical for cancer development. However, little is known about its role and mechanism in ATC. In the present study, the expression of Pol family members is screened in a large-cohort proteome containing 113 ATCs and 20 normal thyroid samples. Combined with the mRNA levels and gene dependency scores, we find that RNA Polymerase I Subunit F (POLR1F) is significantly upregulated in ATC tissues with the strongest gene effect among the Pol family members. The results are confirmed in ATC tissues and cell lines, revealing that POLR1F mainly locates in the nucleus and expresses stronger than that in normal thyrocytes. Silencing POLR1F in ATC cell lines significantly inhibit cell proliferation, colony formation, and sphere sizes. POLR1F knockdown dramatically reduces ATC tumor growth in both zebrafish and nude mouse xenograft models. RNA sequencing reveals that the coagulation factor thrombin receptor (F2R) is a downstream target of POLR1F, which participates in the p38 MAPK pathway. POLR1F promotes the H3K4 methylation at the F2R promoter by reducing the binding of demethylase KDM5C to H3K4me3, thereby enhancing F2R transcription. These results demonstrate that POLR1F maintains ATC stemness and growth by activating F2R/p38 MAPK signaling, shedding light on the essential role of POLR1F in ATC progression.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 5","pages":"Article 119963"},"PeriodicalIF":4.6,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143848578","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"New insights into Gremlin-1: A tumour microenvironment landscape re-engineer and potential therapeutic target","authors":"Chengpeng Sun ,&nbsp;Zijun Ding ,&nbsp;Benjie li ,&nbsp;Sihong Chen ,&nbsp;Enliang Li ,&nbsp;Qingping Yang","doi":"10.1016/j.bbamcr.2025.119962","DOIUrl":"10.1016/j.bbamcr.2025.119962","url":null,"abstract":"<div><div>Gremlin-1 (GREM1), a well-known bone morphogenetic protein (BMP) antagonist, is highly expressed in various malignant tumours. However, the specific role of GREM1 in tumours remains controversial and may be attributed to the heterogeneity and complexity of the tumour microenvironment (TME). It is currently believed that GREM1 regulates the complex landscape of the TME, primarily by antagonising BMP signalling or BMP-independent pathways. Both GREM1 and BMP play dual roles in tumour progression. Therefore, the mutual crosstalk between tumour cells and tumour-associated fibroblasts and the regulation of various secreted factors in the TME affect the secretion level of GREM1, which in turn regulates the amplitude balance between GREM1 and BMP, affecting tumour progression. The inhibition of GREM1 activity in the TME can disrupt this amplitude balance and prevent the formation of a tumour-supportive microenvironment, demonstrating that GREM1 is a potential therapeutic target. In this study, we reviewed the specific signalling pathways via which GREM1 in the TME regulates epithelial-mesenchymal transition, construction of the tumour immune microenvironment, and maintenance of tumour cell stemness via BMP-dependent and BMP-independent regulation, and also summarised the latest clinical progress of GREM1.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 5","pages":"Article 119962"},"PeriodicalIF":4.6,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143848577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In vivo composition of the mitochondrial nucleoid in mice","authors":"Rodolfo García-Villegas ,&nbsp;Franka Odenthal ,&nbsp;Yvonne Giannoula ,&nbsp;Nina A. Bonekamp ,&nbsp;Inge Kühl ,&nbsp;Chan Bae Park ,&nbsp;Henrik Spåhr ,&nbsp;Elisa Motori ,&nbsp;Fredrik Levander ,&nbsp;Nils-Göran Larsson","doi":"10.1016/j.bbamcr.2025.119955","DOIUrl":"10.1016/j.bbamcr.2025.119955","url":null,"abstract":"<div><div>Mitochondrial DNA (mtDNA) is compacted into dynamic structures called mitochondrial nucleoids (mt-nucleoids), with the mitochondrial transcription factor A (TFAM) as the core packaging protein. We generated bacterial artificial chromosome (BAC) transgenic mice expressing FLAG-tagged TFAM protein (<em>Tfam-FLAG</em>^<em>BAC</em> mice) to investigate the mt-nucleoid composition <em>in vivo</em>. Importantly, we show that the TFAM-FLAG protein is functional and complements the absence of the wild-type TFAM protein in homozygous <em>Tfam</em> knockout mice. We performed immunoprecipitation experiments from different mouse tissues and identified 12 proteins as core mt-nucleoid components by proteomics analysis. Among these, eight proteins correspond to mtDNA replication and transcription factors, while the other four are involved in the mitoribosome assembly. In addition, we used the <em>Tfam-FLAG</em>^<em>BAC</em> mice to identify ten proteins that may stabilize TFAM-FLAG upon depletion of the mitochondrial RNA polymerase despite the absence of mtDNA and induction of the LONP1 protease. Finally, we evaluated the changes in mt-nucleoids caused by very high levels of TFAM unraveling nine interactors that could counteract the high TFAM levels to maintain active mtDNA transcription. Altogether, we demonstrate that the <em>Tfam-FLAG</em>^<em>BAC</em> mice are a valuable tool for investigating the mt-nucleoid composition <em>in vivo</em>.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 6","pages":"Article 119955"},"PeriodicalIF":4.6,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143883009","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibition of the Sp1/PI3K/AKT signaling pathway exacerbates doxorubicin-induced cardiomyopathy","authors":"Guojian Xiang ,&nbsp;Tingting Shi ,&nbsp;Cornelius Obinna Nwaele ,&nbsp;Huazhen Xiao ,&nbsp;Yucheng Liu ,&nbsp;Qingfeng Wang ,&nbsp;Jiancheng Zhang ,&nbsp;Yonghong Zheng","doi":"10.1016/j.bbamcr.2025.119960","DOIUrl":"10.1016/j.bbamcr.2025.119960","url":null,"abstract":"<div><h3>Objective</h3><div>This study aimed to investigate the interaction and underlying mechanisms between specificity protein 1 (Sp1) and the phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signaling pathway in the context of doxorubicin-induced cardiomyopathy (DIC).</div></div><div><h3>Methods</h3><div>A rat model of DIC was established by intraperitoneal injection of doxorubicin (1 mg/kg) twice a week for eight weeks. Cardiac function was evaluated using echocardiography, and myocardial histopathology was assessed by hematoxylin-eosin (HE) staining. In vitro, H9c2 cardiomyocytes were treated with doxorubicin (2 μmol/L) to induce cardiotoxicity, followed by co-treatment with the Sp1 inhibitor plicamycin or the PI3K/AKT inhibitor LY294002. Cell viability was measured by the CCK-8 assay. Oxidative stress markers, including reactive oxygen species (ROS) and lactate dehydrogenase (LDH), were quantified using flow cytometry and colorimetric assays. Apoptosis was detected via TUNEL staining, and protein expression of Sp1, PI3K, AKT, and Caspase-3 was analyzed by Western blotting.</div></div><div><h3>Results</h3><div>Doxorubicin treatment significantly impaired cardiac function in rats, as evidenced by an increase in both left ventricular internal diameters during diastole (LVIDd) and systole (LVIDs), along with decreased ejection fraction (EF) and fractional shortening (FS) (<em>p</em> &lt; 0.01). Myocardial HE staining in doxorubicin-treated rats revealed disorganized cardiomyocyte structures, edema, and cellular necrosis. In vitro, doxorubicin exposure led to reduced H9c2 cell viability, elevated ROS and LDH levels, and increased apoptosis rates (<em>p</em> &lt; 0.01). Western blotting demonstrated that doxorubicin significantly downregulated the expression of Sp1, PI3K, and AKT while upregulating Caspase-3. Inhibition of Sp1 or PI3K/AKT exacerbated these effects, resulting in further cardiac dysfunction, oxidative stress, and apoptosis. Moreover, Sp1 inhibition led to decreased PI3K/AKT pathway activation, while PI3K/AKT inhibition reciprocally suppressed Sp1 expression, indicating a bidirectional regulatory relationship.</div></div><div><h3>Conclusion</h3><div>Doxorubicin induces cardiotoxicity by promoting oxidative stress and apoptosis through the downregulation of the Sp1/PI3K/AKT signaling pathway. Inhibition of this pathway exacerbates cardiac injury, suggesting that targeting Sp1 and PI3K/AKT may offer novel therapeutic strategies for the prevention and treatment of DIC.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 5","pages":"Article 119960"},"PeriodicalIF":4.6,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143843877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}