Biocell最新文献

{"title":"The Yin-Yang of Stress and Senescence: Integrated Stress Response and SASP Crosstalk in Stem Cell Fate, Regeneration, and Disease.","authors":"Douglas M Ruden","doi":"10.32604/biocell.2025.072273","DOIUrl":"10.32604/biocell.2025.072273","url":null,"abstract":"<p><p>Stem cell fate decisions are increasingly understood through the dynamic interplay of two fundamental stress-adaptive programs: the integrated stress response (ISR) and the senescence-associated secretory phenotype (SASP). These pathways act as a Yin-Yang system, balancing beneficial and detrimental outcomes across development, tissue homeostasis, and disease. On the yin (protective) side, transient ISR activation and acute SASP signaling foster adaptation, embryonic patterning, wound healing, and regeneration. On the yang (maladaptive) side, chronic ISR signaling and unresolved SASP output drive stem cell exhaustion, fibrosis, inflammation, and tumorigenesis. This duality highlights their roles as both guardians and disruptors of stem cell integrity. Mechanistically, ISR regulates translational control via eukaryotic initiation factor 2 alpha (eIF2α) phosphorylation and activating transcription factor 4 (ATF4)-dependent transcription, while SASP reprograms the extracellular milieu through cytokines, growth factors, and proteases. Their crosstalk creates feedback loops that shape tissue niches and long-term stem cell potential. Framing ISR-SASP interactions through a Yin-Yang lens underscores the balance between resilience and decline, to offer new insights into regenerative medicine, anti-aging interventions, and cancer therapeutics.</p>","PeriodicalId":55384,"journal":{"name":"Biocell","volume":"50 1","pages":""},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12952897/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147349770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Flow Cytometry Study of Immune Cell Subpopulations from the Mouse Vertebral Bone Marrow and Intervertebral Disc Following Endplate Microfracture.","authors":"Dalin Wang, Mingcai Zhang, Richard Hastings, Patrick George, Ryan Ranzau, Jinxi Wang","doi":"10.32604/biocell.2026.074572","DOIUrl":"10.32604/biocell.2026.074572","url":null,"abstract":"<p><strong>Objective: </strong>Although endplate (EP) injury may cause intervertebral disc (IVD) degeneration and Modic changes (MCs) in the vertebral bone marrow (VBM), EP injury-induced synchronous cellular reactions and their crosstalk in the IVD and VBM remain unclear. This protocol-based study aimed to streamline and optimize the methods of tissue harvest and cell preparation for flow cytometry (FCM) analysis of T-cell and macrophage subpopulations in both VBM and IVD adjacent to the surgically induced EP microfracture in mice.</p><p><strong>Methods: </strong>EP injury or sham procedure was performed at the spinal levels L4-5 and L5-6 in male mice. Step-by-step techniques on the harvest of lumbar VBM and IVD tissues, isolation of tissue-specific cells, and generation of single cell suspensions were described. FCM analysis was performed using specific antibodies against cell-surface markers and multi-color cytometry for precise delineation of cell subsets to explore the cellular mechanism of MCs. An extracellular staining assay to identify macrophage subsets, as well as extra- and intra-cellular staining assays to identify T lymphocyte subsets from the VBM and IVD, were performed and optimized.</p><p><strong>Results: </strong>FCM analysis demonstrated that significantly increased macrophage numbers and M2a polarization response were observed in VBM samples from the sham and EP injury groups, while the ratio of M2a/total number of macrophages was significantly increased and the ratio of M1/M2a was significantly decreased in IVDs from the sham and EP injury groups. A significantly increased Treg cell response was detected in VBM samples from the EP injury group, but not the sham group.</p><p><strong>Conclusions: </strong>This protocol reports novel and reproducible methods of tissue harvest, cell preparation, and antibody selection for flow cytometry analysis of T-cell and macrophage subpopulations isolated from the VBM and IVD following EP injury in mice. This protocol may be utilized for exploring the cellular mechanism of MCs and IVD degeneration in animal models.</p>","PeriodicalId":55384,"journal":{"name":"Biocell","volume":"50 4","pages":""},"PeriodicalIF":1.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13337323/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148438925","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Emerging Roles of Fc Receptor-Like 1 in Immunotherapy of Diffuse Large B-Cell Lymphoma.","authors":"Kayce Blumenstock, Vandana Zaman, Camille Green, Narendra L Banik, Azizul Haque","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Fc Receptor-Like 1 (FCRL1), a member of the FCRL family, contains two immunoreceptor tyrosine-based activation motifs (ITAMs) in its cytoplasmic domain and plays a critical role in B-cell biology. Its expression begins in pre-B-cells, dynamically shifts during B-cell development, and contributes to the regulation of human B-cell activation. Notably, FCRL1 is overexpressed in subsets of naive and memory B-cells, as well as in malignant B-cells, including those in diffuse large B-cell lymphoma (DLBCL), an aggressive and often treatment-resistant hematological malignancy. Among FCRL family members, FCRL1 stands out as a promising immunotherapeutic target due to its selective expression in malignant B-cells and its functional role in proliferation. Given the limited efficacy of current therapies for relapsed/refractory DLBCL, targeting FCRL1 could address an unmet clinical need by offering a novel, mechanism-based approach to modulate B-cell signaling and enhance anti-tumor immunity. This mini-review highlights the therapeutic potential of FCRL1-directed strategies, supporting their further exploration in preclinical models and future clinical trials for DLBCL and other B-cell malignancies.</p>","PeriodicalId":55384,"journal":{"name":"Biocell","volume":"49 10","pages":"1859-1871"},"PeriodicalIF":1.8,"publicationDate":"2025-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12981233/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147470350","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"miR-122-5p Regulates Ferroptosis through Targeting the Glutamine Transporter SLC1A5 in Hepatocellular Carcinoma","authors":"Mei G. Lei, Jingli Xu, Xiaoying Hu, Feng Lin, Baoping Luo","doi":"10.32604/biocell.2025.068926","DOIUrl":"https://doi.org/10.32604/biocell.2025.068926","url":null,"abstract":"Background: Hepatocellular carcinoma (HCC) typically begins inconspicuously and progresses swiftly, leading to most patients being diagnosed at an advanced stage. Accordingly, a pressing priority is to clarify the development mechanisms of HCC and devise efficient intervention and treatment protocols. Methods: An upstream miRNA of solute carrier transporter family 1 member 5 (SLC1A5) was predicted to be miR-122-5p by various databases, and a dual-luciferase reporter gene assay was used to verify the SLC1A5- and miR-122-5p-targeting relationship. SLC1A5 and miR-122-5p expression in HCC cells was quantitatively assessed using quantitative reverse transcription polymerase chain reaction (qRT–PCR). Western blotting was used to evaluate SLC1A5 expression in HCC cells. To determine the effects of the ferroptosis inducers erastin and L-g-glutamyl-p-nitroanilide (GPNA) on Huh-7 and HepG2 cell viability, a Cell Counting Kit-8 assay was performed. Additionally, various assay kits were used to assess malondialdehyde (MDA), Fe2+, and reactive oxygen species (ROS) levels in HCC cells. Moreover, an HCC tumor model was established in nude mice to investigate the growth of HCC tumors overexpressing miR-122-5p. Results: miR-122-5p downregulated SLC1A5 levels. MiR-122-5p knockdown inhibited erastin-promoted ferroptosis, whereas miR-122-5p overexpression promoted ferroptosis. After knocking down miR-122-5p, the MDA, Fe2+, and ROS levels in Huh-7 and HepG2 cells decreased, whereas overexpressing miR-122-5p increased the MDA, Fe2+, and ROS levels. Following the addition of GPNA, the cells experienced decreased viability and increased MDA levels, which in turn inhibited ferroptosis. Knockdown of SLC1A5 partially reversed the ferroptosis-inhibiting effect induced by knocking down miR-122-5p. Additionally, the overexpression of miR-122-5p hindered HCC development in vivo. Conclusion: miR-122-5p downregulates SLC1A5, which promotes lipid peroxidation and iron accumulation and inhibits glutamine transport, ultimately causing ferroptosis in HCC cells.","PeriodicalId":55384,"journal":{"name":"Biocell","volume":"0 0","pages":"1-10"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147334116","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrin Alpha8 Beta1 (81): An In-Depth Review of an Overlooked RGD-Binding Receptor.","authors":"Iman Ezzat, Marisa Zallocchi","doi":"10.32604/biocell.2025.062325","DOIUrl":"10.32604/biocell.2025.062325","url":null,"abstract":"<p><p>Integrins are heterodimeric transmembrane receptors that mediate bidirectional interactions between the intracellular cytoskeletal array and the extracellular matrix. These interactions are critical in tissue development and function by regulating gene expression and sustaining tissue architecture. In humans, the integrin family is composed of 18 alpha (α) and 8 beta (β) subunits, constituting 24 distinct αβ combinations. Based on their structure and ligand-binding properties, only a subset of integrins, 8 out of 24, recognizes the arginine-glycine-aspartate (RGD) tripeptide motif in the native ligand. One of the major RGD binding integrins is integrin alpha 8 beta 1 (α8β1), a central Ras homolog gene family member A (RHOA)-dependent modulator highly expressed in cells with contractile function. This review focuses on the recent advances regarding α8β1 function during organ development, with a particular interest in kidney and inner ear development. We also discuss α8β1's role in injury and disease and its importance for mesenchymal to epithelial transition during cancer development. Finally, we highlight α8β1's importance for hearing function and its future use as a potential diagnostic and therapeutic tool for disease elimination.</p>","PeriodicalId":55384,"journal":{"name":"Biocell","volume":"49 5","pages":"789-811"},"PeriodicalIF":0.8,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12162094/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144287165","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"ScRNA-seq and Experimental Analyses Unveil Lrg1 Regulating the Oxidative Phosphorylation Pathway to Affect Neutrophil Accumulation after Cerebral Ischemia-Reperfusion","authors":"Luyao Jiang, Longsheng Fu, Shaofeng Xiong, Guosheng Cao, Yanqin Mei, Yaoqi Wu, Jin Chen, YANNI LV","doi":"10.32604/biocell.2025.068507","DOIUrl":"https://doi.org/10.32604/biocell.2025.068507","url":null,"abstract":"Background: After ischemic stroke, neutrophils hyperactivate, increasing in number and worsening inflammation, causing neural damage. Prior scRNA-seq showed Lrg1 modulates cells subsentence to cerebral ischemia-reperfusion injury, but its mechanism in regulating neutrophil accumulation/differentiation post-injury is unclear. Methods: Lrg1 knockout impact on neutrophil accumulation was assessed via immunofluorescence and western blot. Three-dimensional reconstruction of immunofluorescent staining analyzed cell-cell interactions among neutrophils and microglia. scRNA-seq of WT and Lrg1-/- mice from GSE245386 and GSE279462 was conducted. Each group conducted oxidative phosphorylation scoring via Gene Set Enrichment Analysis (GSEA), while Metascape was employed to perform GO and KEGG enrichment analyses for elucidating functional mechanisms. CellChat exhibited cell-cell communication. Furthermore, alterations in microglial phagocytic activity were evaluated by immunostaining for CD68, a well-established marker of phagolysosomal activity in phagocytic cells. Brain energy metabolism was evaluated via glutamate dehydrogenase activity and ATP levels with ELISA, and enzyme expression was analyzed by immunofluorescence and western blot. Results: Lrg1 knockout decreased neutrophil accumulation and NET formation in mice. 3D immunofluorescence reconstruction confirmed neutrophil co-localization with endothelial cells/microglia. scRNA-seq revealed that the oxidative phosphorylation score was significantly higher in the MCAO/R+WT group compared to both the Sham-operated+WT and Lrg1-/- groups. Notably, the oxidative phosphorylation score was further elevated in the MCAO/R+Lrg1-/- group. Immunostaining showed that Lrg1 knockout elevated CD68+ lysosome expression post-MCAO/R, with TMEM119 colocalizing with these lysosomes. MCAO/R raised CD68 expression in ischemic brains, an effect further intensified by Lrg1 knockout. KEGG analysis linked differential genes to oxidative phosphorylation pathways. Validation in MCAO/R vs. sham groups revealed increased ROS production and reduced expression of complex enzymes I-V (NDUFB8, SDHB, UQCRC1, MTCO2, ATP5A1). Lrg1 intervention increased enzyme expression. Immunofluorescence and western blot in brain tissue showed similar patterns in microglia and enzymes I-V. Conclusions: Lrg1 knockout significantly enhances microglial phagocytic activity towards neutrophils subsequent to cerebral ischemia-reperfusion injury, through its regulatory effect on the oxidative phosphorylation pathway. This finding accentuates Lrg1 as a highly potential therapeutic target for intervening in and modulating post-ischemic inflammatory responses.","PeriodicalId":55384,"journal":{"name":"Biocell","volume":"49 9","pages":"1749-1769"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147882405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rhein Inhibits Podocyte Ferroptosis and Epithelial-Mesenchymal Transition in Diabetic Nephropathy by Activating the SIRT1/p53/SLC7A11 Pathway","authors":"Wei‐Syun Hu, Dan Xiong","doi":"10.32604/biocell.2025.067670","DOIUrl":"https://doi.org/10.32604/biocell.2025.067670","url":null,"abstract":"Objectives: Podocytes undergo epithelial-mesenchymal transition (EMT) and ferroptosis in response to hyperglycemic stimulation. This is considered an important early event in the development and progression of diabetic nephropathy (DN). Rhein is the main active anthraquinone derivative in several common traditional herbal medicines. This study aimed to investigate the protective effects of Rhein on podocyte ferroptosis and EMT. Methods: The mouse glomerular podocyte cell line MPC5 was stimulated with high glucose (HG), Rhein, and the ferroptosis inhibitor ferrostatin-1 (Fer-1). Mechanistic investigations employed plasmids to overexpress and knockdown Sirtuin-1 (SIRT1), solute carrier family 7 member 11 (SLC7A11), or p53 and measure ferroptosis- or EMT-related indicators. Results: In the HG-injured podocytes, Rhein enhanced cell viability, reduced malondialdehyde (MDA), ferrous iron (Fe2+), and reactive oxygen species (ROS) levels, increased glutathione (GSH) production, accompanied by the restoration of ferroptosis- and EMT-associated indicator expressions. Mechanistically, Rhein induced SIRT1 and SLC7A11 expression and attenuated p53 expression. SIRT1 knockdown upregulated p53 and downregulated SLC7A11, thereby abolishing the protective effects of Rhein against podocyte ferroptosis and EMT. However, the effects of SIRT1 overexpression were reversed by SLC7A11 knockdown. Conclusion: Rhein activated the SIRT1/p53/SLC7A11 axis to protect podocytes against ferroptosis and EMT. This suggests that Rhein has a potential therapeutic effect on DN patients associated with podocyte injury, and targeting SIRT1/p53/SLC7A11 may represent an innovative therapeutic strategy for DN patients.","PeriodicalId":55384,"journal":{"name":"Biocell","volume":"49 9","pages":"1711-1731"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147893411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stretch Enhances Secretion of Extracellular Vehicles from Airway Smooth Muscle Cells via Endoplasmic Reticulum Stress Signaling in Relation to Ventilator-Induced Lung Injury","authors":"Mingzhi Luo, CHANGYU SUN, Jia Guo, Xiangrong Zhang, Jing Zhang, XUANYU SHI, Lei Liu, Yan Pan, Jingjing Li, Linhong Deng","doi":"10.32604/biocell.2025.063869","DOIUrl":"https://doi.org/10.32604/biocell.2025.063869","url":null,"abstract":"Background: Mechanical ventilation (MV) provides life support for patients with severe respiratory distress but can simultaneously cause ventilator-induced lung injury (VILI). However, due to a poor understanding of its mechanism, there is still a lack of effective remedies for the often-deadly VILI. Recent studies indicate that the stretch associated with MV can enhance the secretion of extracellular vesicles (EVs) and induce endoplasmic reticulum (ER) stress in airway smooth muscle cells (ASMCs), both of which can contribute to VILI. But whether MV-associated stretch enhances the secretion of EVs via ER stress in ASMCs as an underlying mechanism of VILI remains unknown. Methods: In this study, we exposed cultured human ASMCs to stretch (13% strain) and mouse models to MV at tidal volume (18 mL/kg). Subsequently, the amount of secreted EVs in the culture medium of ASMCs and the bronchoalveolar lavage fluid (BALF) of mouse models was quantitatively evaluated by ultracentrifugation, transmission electron microscopy, Western blot, flow cytometry, and nanoparticle tracking analysis. The cultured ASMCs and the lung tissues of mouse models were assessed for expression of biomarkers of EVs (cluster of differentiation antigen 63, CD63), ER stress (heat shock protein family A member 5, HSPA5), and EVs regulating molecule Rab27a by immunofluorescence microscopy, immunohistochemistry (IHC) and enzyme-linked immunosorbent assay (ELISA), respectively. MicroRNAs (miRNAs) in EVs from ASMCs were measured with miRNA whole genome sequencing (miRNA-Seq). Results: We found that stretch enhanced EV secretion from cultured ASMCs. In addition, the cultured ASMCs and the mouse models were either or not pretreated with ER stress inhibitor (tauroursodeoxycholic acid, TUDCA)/EV secretion inhibitor (GW4869) prior to stretch or MV. We found that MV-associated stretch enhanced the expression of CD63, HSPA5, and Rab27a in cultured ASMCs and BALF/lung tissues of mouse models, which could all be attenuated with TUDCA/GW4869 pretreatment. miRNA-Seq data show that differentially expressed miRNAs in EVs mainly modulate gene transcription. Furthermore, the EVs from cultured ASMCs under stretch tended to enhance detachment and expression of inflammatory cytokines, i.e., transforming growth factor-β1 (TGF-β1), interleukin-10 (IL-10) in cultured airway epithelial cells. The expression of TGF-β1 and IL-10 in BALF of the mouse models also increased in response to MV, which was attenuated together with partial improvement of lung injury by pretreatment with TUDCA, GW4869/Rab27a siRNAs. Conclusion: Taken together, our data indicate that MV-associated stretch can enhance the secretion of EVs from ASMCs via ER stress signaling to mediate airway inflammation and VILI, which provides new insight for further exploring EVs for the diagnosis and treatment of VILI.","PeriodicalId":55384,"journal":{"name":"Biocell","volume":"49 5","pages":"833-855"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147911017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"&lt;i&gt;TRIM32&lt;/i&gt; Promotes Glycolysis in Keloid Fibroblasts and Progression of Keloid Scars via Regulation of the PI3K/AKT Signaling Pathway","authors":"Yi Zhang, Hua Jin","doi":"10.32604/biocell.2025.066479","DOIUrl":"https://doi.org/10.32604/biocell.2025.066479","url":null,"abstract":"Objectives: The present study investigated whether Tripartite Motif-Containing Protein 32 (TRIM32) contributes to the aberrant activation of keloid fibroblasts (KFs) via glycolysis. Methods: The expression levels of TRIM32, ... | Find, read and cite all the research you need on Tech Science Press","PeriodicalId":55384,"journal":{"name":"Biocell","volume":"49 8","pages":"1529-1543"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147918412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeting EphA2 with CRISPR/Cas9 Identifies a Novel EphA2-CDH1 Regulatory Axis in Migration of Castration-Resistant Prostate Cancer","authors":"Yan Song, Yi-Ting Chen, Xiaojing Li, Peng Pan, Nan Hu, Lanyi Wei, Yue Xiao, Chaogang Wei","doi":"10.32604/biocell.2025.065658","DOIUrl":"https://doi.org/10.32604/biocell.2025.065658","url":null,"abstract":"Objectives: Progression to castration-resistant prostate cancer (CRPC) and metastasis are the greatest challenges to effective treatment. Anticancer strategies targeting the key kinases associated with the development of CRPC may represent a breakthrough. The tyrosine kinase receptor Erythropoietin-producing hepatocellular (Eph) A2 receptor is highly expressed in CRPC cell lines and may be associated with tumor invasion and metastasis. However, the effects and exact mechanisms of EphA2 in CRPC are only partially understood. This study aimed to investigate the impact of EphA2 on CRPC cell behaviors and underlying molecular pathways Methods: CRISPR/Cas9-mediated gene editing induced EphA2-disrupted in human-derived PC3 and DU145 cells. Single-guide RNAs (sgRNAs) targeting EphA2 were designed, and editing efficiency was validated. Optimal sgRNA sequences were selected to generate EphA2-knockdown (KD) and -overexpressing (OE) cell lines. Cell migration, proliferation, and apoptosis were assessed via functional assays. Transcriptomic analysis, quantitative PCR, and Western blotting were performed to identify downstream effectors. Bioinformatics analyses were used to correlate EphA2 and CDH1 expression with clinical parameters in prostate cancer patients. Results: Editing efficiency was found to vary among different sgRNAs targeting the EphA2 gene. EphA2-KD significantly inhibited CRPC cell migration but did not affect cell proliferation or apoptosis. Conversely, EphA2-OE significantly enhanced the migration of DU145 cells. Molecular analyses revealed that the expression of CDH1 (an important marker of the epithelial-mesenchymal transition (EMT) in tumors) was significantly upregulated in PC3-EphA2-KD cells and downregulated in DU145-EphA2-OE cells, indicating that CDH1 is a downstream regulator of EphA2. Bioinformatic analysis revealed that higher EphA2 levels and lower CDH1 expression were both associated with an advanced tumor T stage, higher Gleason scores, and lymph node metastases in prostate cancer patients. More importantly, EphA2 was found to be an important predictor of lymph node metastasis, in addition to the Gleason score. Adding EphA2 to the Gleason score could significantly improve the detection of lymph node metastasis. Conclusion: CRISPR/Cas9-mediated EphA2-KD significantly suppressed the migration of CRPC cells through the inhibition of the EphA2-CDH1 axis. Strategies targeting the EphA2 gene may be promising for the treatment of CRPC.","PeriodicalId":55384,"journal":{"name":"Biocell","volume":"49 7","pages":"1207-1223"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147883183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}