OBJECTIVE:Cancer-associated fibroblasts (CAFs) contribute to pancreatic cancer (PC) progression and therapeutic resistance. This study investigated whether CAF-derived exosomes (CAF-exo)-promote PC cell stemness and gemcitabine (GEM) resistance by delivering Snail and regulating the suppressor of cytokine signaling 6/hypoxia-inducible factor-1α (SOCS6/HIF-1α) axis. METHODS:Tumors and paired adjacent tissues from patients with PC were collected. RT-qPCR measured Snail, SOCS6, and HIF-1α mRNA; correlations were assessed by Pearson analysis, and survival by Kaplan-Meier. CAFs and normal fibroblasts were isolated for Exo extraction. PC cells (SW1990/MiaPACa2) were treated with lentiviral sh-Snail-transfected CAF-exo, oe-Snail, oe-SOCS6, or si-HIF-1α. Nanog, octamer-binding transcription factor 4 (OCT4), SRY-box transcription factor 2 (SOX2), Snail, SOCS6, and HIF-1α, as well as spheroid formation, chemoresistance, viability, and apoptosis, were evaluated. Snail's binding to the SOCS6 promoter was confirmed by dual-luciferase and ChIP-qPCR. A xenograft mouse model was established for in vivo validation. RESULTS:In PC tissues, Snail and HIF-1α were elevated, SOCS6 reduced; Snail negatively correlated with SOCS6 and positively with HIF-1α, and these patterns were associated with poor survival. CAF-exo delivered Snail into PC cells, elevating stemness markers, sphere formation, and GEM resistance, which were reversed by Snail knockdown. Mechanistically, CAF-exo transcriptionally represses SOCS6 via Snail, leading to HIF-1α upregulation. SOCS6 upregulation or HIF-1α knockdown partly attenuated CAF-exo-induced stemness and GEM resistance. In vivo, CAF-exo promoted tumor growth, stemness, and GEM resistance and reduced apoptosis via the Snail-SOCS6/HIF-1α axis. CONCLUSION:CAF-exo shuttled Snail, transcriptionally suppressed SOCS6 expression to activate HIF-1α, thereby enhancing PC cell stemness and GEM resistance in PC cells.
BACKGROUND:Liver fibrosis (LF) lacks effective therapies. Longevity assurance homolog 2 (LASS2), a key ceramide synthase, contributes to liver homeostasis, but its role in LF remain unclear. METHODS:Bioinformatics revealed LASS2 correlation with classic fibrotic factors, and its expression was assessed in fibrotic mouse livers and activated LX-2 cells. Gain- and loss-of-function studies were performed in vivo (liver-specific AAV) and in vitro (recombinant adenovirus), followed by phenotypic and mechanistic analyses. Protein interactions were mapped by proteomics and co-immunoprecipitation (co-IP), and direct interactions were validated by protein-protein docking, co-IP/Western blot, immunofluorescence colocalization, and proximity ligation assay (PLA). Hepatocyte conditioned medium (CM) experiments combined with ELISA and targeted lipidomics were performed to dissect LASS2-mediated paracrine signaling. RESULTS:LASS2 overexpression attenuated fibrogenesis, regulated mtROS in an activation-dependent manner, promoted apoptosis in both quiescent and activated hepatic stellate cells (HSCs), and suppressed EMT. Mechanistically, LASS2 directly interacted with thrombospondin 1 (THBS1), transferrin receptor (TFRC), and apolipoprotein E (APOE). These interactions correlated with inhibition of TGF-β1/Smad2/3 signaling, remodeled lipid metabolism, coordinately suppresses ferroptosis and promotes apoptosis in activated HSCs, thereby attenuating liver fibrosis. Notably, LASS2 overexpression in hepatocytes reduced CM levels of TGF-β1 and induced global lipidomic remodeling, characterized by downregulation of pro-fibrotic lysophospholipids (LPC/LPE/LPI), phosphatidic acid (PA), and ceramides (Cer), alongside upregulation of specific sphingomyelin (SM) and BMP species, creating an anti-fibrotic paracrine milieu. CONCLUSIONS:LASS2 acts as a dual-function metabolic-signaling integrator: it directly engages THBS1, TFRC, and APOE to control TGF-β1 signaling and ferroptosis, while also remodeling the hepatocyte-derived paracrine lipid landscape to inhibit HSC activation. Targeting LASS2 thus represents a promising anti-fibrotic strategy.
Breast cancer (BC) is a notoriously difficult malignancy to treat due to its high molecular heterogeneity and drug resistance. The vast structural diversity of marine-derived compounds remains underutilized in oncology drug development. This study investigates the anti-tumor potential and underlying molecular mechanism of cycloechinulin (CCL), a novel diketopiperazine alkaloid isolated from the marine-derived fungus Aspergillus ochraceus against BC. The anti-cancer efficacy of CCL was evaluated using in vitro phenotypic assays in BC cell lines and validated in patient-derived organoids (PDOs). Global transcriptional changes were profiled via RNA sequencing (RNA-seq), and the clinical significance of downstream targets was analyzed using public patient databases. CCL significantly reduced BC cell viability, suppressed DNA replication, and restricted migration and invasion while inducing cellular apoptosis. CCL treatment also led to robust growth inhibition and morphological degradation across multiple patient-derived breast cancer models. Transcriptomic profiling and bioinformatic validation identified Shugoshin 2 (SGOL2) as a key oncogenic biomarker significantly correlated with advanced clinical stages and poor patient survival. At the molecular level, CCL downregulated the protein expression of SGOL2 as well as its downstream effectors METTL3 and ICAM2. Furthermore, knockdown of SGOL2 reproduced CCL-mediated anti-tumor effects and suppressed METTL3 and ICAM2 expression. In conclusion, CCL suppresses breast cancer progression by inhibiting the novel SGOL2/METTL3/ICAM2 signaling axis. These results present CCL as a promising natural lead compound and a novel chemical scaffold for targeted BC therapeutics.
BACKGROUND:Alzheimer's disease (AD) is the most common neurodegenerative disorder worldwide and the leading cause of dementia. Mitochondrial dysfunction is a key pathogenic event that drives neuronal damage and disease progression. Thus, protecting against mitochondrial damage in neuronal cells has become a critical therapeutic target for AD prevention and treatment. Our previous research has shown that artemether can protect PC12 cells from oxidative stress damage induced by Aβ and oxygen-glucose deprivation, but its underlying regulatory mechanisms remain elusive. Therefore, it is essential to elucidate the role and mechanism of artemether in Aβ-induced mitochondrial damage. OBJECTIVE:Evaluate the protective effects of artemether on Aβ-induced mitochondrial dysfunction and the underlying molecular mechanism in AD. METHODS:The HT-22 cell line and primary hippocampal neurons were employed in the present study owing to their greater physiological relevance to hippocampal neuronal injury. In order to evaluate the protective effects of artemether against mitochondrial damage in neuronal cells, we performed Cell Counting Kit-8 assays to assess cell viability and LDH release assays to measure cytotoxicity. Furthermore, we evaluated mitochondrial membrane potential, quantified ATP content, and detected ROS production in Aβ-induced neuronal cell damage. In addition, we used transmission electron microscopy to observe mitochondrial ultrastructural changes and performed immunofluorescence staining to analyze mitochondrial protein localization and morphology. In the AD mouse model, we conducted behavioral tests to assess cognitive function, while using H&E staining, immunohistochemistry, and TUNEL staining to evaluate artemether's effects on AD-related pathological hallmarks. Furthermore, Western blot analysis was performed to dissect the molecular mechanism underlying artemether's protective effect against Aβ-induced mitochondrial damage, focusing on the expression of key proteins in the PGC1/ERRα/TFAM signaling pathway. RESULTS:Artemether protects against Aβ1-42 induced mitochondrial damage in HT-22 cells and alleviates Aβ1-42 injection induced memory deficits, mitochondrial dysfunction and neuroinflammation in the AD mice model. Mechanistically, artemether treatment upregulated the protein levels of PGC1, which contributes to mitochondrial homeostasis and further activation of the ERRα/TFAM signaling pathway. Moreover, Inhibition of PGC1α abrogates artemether's protective effects. CONCLUSION:Our findings indicate that neuronal mitochondrial dysfunction serves as a central driver in AD pathogenesis, and activating the PGC1α/ERRα/TFAM axis through artemether offers an effective and feasible strategy for the prevention and treatment of AD.
Activation of microglia contributes to the pathogenesis of central neuropathic pain (CNP), yet effective treatments remain limited. Docosahexaenoyl-ethanolamine (DHEA) and eicosapentaenoyl-ethanolamine (EPEA) are omega-3-derived ethanolamides with reported immunoregulatory and neuroprotective actions. We examined the effects of DHEA and EPEA, alone and in combination, on microglial activation, microglia-neuron crosstalk, and cannabinoid receptor-associated signalling. In LPS-activated microglia, DHEA and EPEA reduced pro-inflammatory mediator expression, attenuated inflammatory cytokine and chemokine profiles, and increased anti-inflammatory mediators including BMP7. Conditioned medium from activated microglia induced neuronal stress, whereas conditioned medium from DHEA-treated microglia, as well as BMP7 alone, attenuated this effect. DHEA and EPEA promoted via CB1R- and CB2R-dependent Gαi and β-arrestin interaction, receptor internalisation, ERK activation, and reduced cAMP accumulation. Together, these findings show that DHEA and EPEA suppress microglial inflammatory signalling and reduce microglia-induced neuronal stress, support a role for CBR-receptor signalling in their actions and provide evidence for their therapeutic potential in central neuropathic pain.
JMJD2D is a histone demethylase implicated in cancer-associated gene regulation, but its role in non-small cell lung cancer (NSCLC) remains incompletely defined. We examined JMJD2D expression and functional effects in patient specimens and lung cancer models. JMJD2D expression was higher in 41 paired NSCLC tissues than in adjacent non-tumor tissues, was greater in stage III-IV than in stage I-II tumors, and was associated with worse overall survival. JMJD2D was also elevated in A549 and H1975 cells relative to HBE cells. shRNA-mediated JMJD2D knockdown reduced CCK-8 and colony-formation readouts and attenuated migration and invasion in both cell lines; it also reduced xenograft tumor growth and lowered MYC, CCND1, MMP2, and MMP9 mRNA and protein levels in tumors. In A549 cells, endogenous co-immunoprecipitation detected JMJD2D and β-catenin in the same complex. PRI-724 or Z-JIB-04 treatment conditions were associated with lower malignancy-associated readouts and lower levels of selected transcripts, without an apparent change in total β-catenin abundance. These findings associate JMJD2D with aggressive clinical and experimental phenotypes in lung cancer and support further investigation of its molecular role and therapeutic relevance.
Interferon regulatory factor 4 (IRF4) is a lineage-restricted transcription factor essential for plasma cell differentiation and represents an important dependency in multiple myeloma (MM). Unlike other IRF family members, IRF4 is primarily induced by antigen receptor signaling rather than interferon stimulation, linking B-cell development, plasma cell biology, and MM pathogenesis. In MM, IRF4 expression and activity are regulated by multilayered genetic, epigenetic, transcriptional, post-transcriptional, and post-translational mechanisms, collectively maintaining IRF4-centered transcriptional programs that support malignant plasma cell identity and survival. Functionally, IRF4 regulates transcriptional networks controlling myeloma cell proliferation, stress adaptation, and survival, while emerging evidence indicates that IRF4 may also have context-dependent roles in immune regulation. The dependency of malignant plasma cells on IRF4 provides a rationale for therapeutic targeting, and multiple strategies have been developed to modulate IRF4 expression or activity. This review provides an integrated overview of IRF4 regulation and function in MM, highlighting its role in disease pathogenesis, therapeutic strategies, translational challenges, and future opportunities for precision targeting.
OBJECTIVE:To investigate the role of Src tyrosine kinase in Zn2+-induced cardioprotection and the mechanisms involving endoplasmic reticulum stress (ERS), mitochondrial dysfunction, and mitochondrial calcium regulation. METHODS:H9c2 cells were used to establish in vitro models of ischemia/reperfusion (I/R) injury and ERS. Cell viability and cytotoxicity were assessed using MTT/CCK-8 assays and lactate dehydrogenase release. Co-immunoprecipitation was performed to examine the interactions of Src with MCU and Mfn1/Mfn2. Western blotting was used to detect Src phosphorylation, MCU complex components, CaMKIIδ/CREB signaling proteins, and ERS markers. Confocal microscopy was used to evaluate mitochondrial membrane potential, Zn2+ and Ca2+ levels, and reactive oxygen species generation. RESULTS:In 2-DG/TM-treated H9c2 cells, GRP78/94 expression was increased, cell viability was reduced, Src phosphorylation was suppressed, and intracellular and mitochondrial Zn2+ levels were decreased. These changes were accompanied by mitochondrial membrane potential loss, mPTP opening, Ca2+ overload, and ROS accumulation. Exogenous Zn2+ reversed these effects, whereas PP2 or Src siRNA reversed its protection. In the I/R model, Zn2+ improved cell viability, reduced LDH release, restored Zn2+ homeostasis, and alleviated Ca2+ overload. I/R decreased p-Src (Tyr416) and MCUb expression while increasing CaMKIIδ, MCU, MICU1, MICU2, and p-CREB (Ser133). These alterations were reversed by Zn2+ and attenuated by PP2. CONCLUSIONS:Zn2+ protects H9c2 cells against ERS and I/R injury through a Src-dependent mechanism. Activated Src is associated with Mfn1/Mfn2 and preserves mitochondrial function during ERS. In addition, Zn2+ regulates the CaMKIIδ/CREB/MCU pathway through Src activation, thereby reducing mitochondrial Ca2+ overload, mPTP opening, ROS accumulation, and I/R-induced cardiomyocyte injury.
Persistent activation of the Notch signaling pathway is closely associated with the maintenance of cancer stem cells (CSCs) and the progression of hepatocellular carcinoma (HCC). Recombination signal binding protein for immunoglobulin kappa J region (RBPJ), a key transcription factor that mediates downstream target gene transcription in the Notch pathway, has not been fully characterized with respect to its role in promoting CSC traits in HCC. In this study, we found that RBPJ is markedly upregulated in HCC and is associated with selected clinicopathologic features. RBPJ also promoted stem-like phenotypes in HCC cells. Mechanistically, TRIM41 interacts with RBPJ and mediates K63-linked polyubiquitination of RBPJ at Lys135, thereby stabilizing RBPJ and enhancing RBPJ-mediated transcriptional activation of the stemness-related genes SOX2 and CD44. Consequently, the TRIM41-RBPJ axis sustains CSC-like properties in HCC cells. Our findings identify a previously unrecognized post-translational mechanism by which TRIM41-mediated stabilization of RBPJ promotes stem-like properties. These findings expand our understanding of the post-translational regulation of RBPJ in HCC and highlight the TRIM41-RBPJ axis as a potential molecular target warranting further investigation.
BACKGROUND:Lipophagy-driven metabolic reprogramming is increasingly recognized as a determinant of tumor progression. TRIB3, a key regulator of lipid metabolism, has been implicated in cancer aggressiveness, with elevated expression linked to poor outcomes in triple-negative breast cancer. Here, we investigated whether TRIB3 promotes TNBC metastasis through regulation of lipophagy and lipid metabolism. METHODS:Public datasets (Oncomine, GEPIA, Kaplan-Meier Plotter) were analyzed to assess TRIB3 expression and its prognostic relevance in breast cancer. Immunohistochemistry and Western blotting validated TRIB3 expression in TNBC tissues and adjacent normal samples. Functional assays, including wound healing, Transwell migration, and EMT marker detection, were used to determine the effects of TRIB3 on cell migration and invasion. Subcutaneous xenograft and lung metastasis models were established in nude mice to evaluate its role in vivo. Potential upstream regulators were identified by dual-luciferase reporter assays, while downstream pathways were investigated using RNA sequencing, pathway enrichment, and functional annotation. The impact of TRIB3 on lipophagy was examined by transmission electron microscopy, immunofluorescence, and Western blotting, and further validated using autophagy inhibitors. RESULTS:TRIB3 was significantly overexpressed in TNBC and correlated with unfavourable prognosis. Functional assays demonstrated that TRIB3 enhanced TNBC cell invasion and metastasis both in vitro and in vivo. Mechanistically, TRIB3 negatively regulated MSI2 protein expression and activated the PI3K/AKT/mTOR pathway, thereby disrupting lipid droplet redistribution and inhibiting lipophagy, which facilitated tumor migration and metastasis. CONCLUSIONS:Our findings uncover a previously unrecognized mechanism by which TRIB3 regulates lipophagy to drive TNBC metastasis. We delineate the critical role of the TRIB3-MSI2 regulatory axis and its association with the PI3K/AKT/mTOR pathway, as well as the potential upstream involvement of FOXA1, in orchestrating this process. These results highlight TRIB3 as a central mediator of TNBC progression and suggest novel therapeutic strategies targeting lipophagy for this aggressive breast cancer subtype.
BACKGROUND:Colorectal cancer (CRC) progression is linked to aberrant cellular senescence. Netrin-1/UNC5B signaling regulates tumor cell survival, yet whether indole-3-carboxylic acid (I3C) modulates CRC senescence via this pathway remains unknown. This study aims to investigate I3C's mechanism in CRC senescence and its connection to Netrin-1/UNC5B signaling. METHODS:HCT-116 and HT29 cells were exposed to I3C. Senescence was assessed and the targeted binding relationship and key binding sites between I3C and UNC5B were verified. UNC5B knockdown/mutant cell lines were established; combined with intervention using exogenous recombinant Netrin-1 and Wnt/β-catenin inhibitor XAV939, downstream mechanism of Netrin-1/UNC5B pathway was clarified. A xenograft model was constructed to validate the anti-tumor effect and molecular targets of I3C. RESULTS:I3C induced CRC cell senescence concentration- and time-dependently. I3C specifically bound UNC5B residues (SER604, THR590, GLU718), inhibiting Netrin-1/UNC5B interaction, with SER604 identified as a functionally critical regulatory residue. Notably, loss-of-function of UNC5B attenuated I3C's pro-senescence effect. Netrin-1/UNC5B signaling could activate Wnt/β-catenin pathway (upregulating Axin2, c-Myc, Cyclin D1), counteracting doxorubicin-induced senescence, while XAV939 reversed this activation. Functional assays confirmed that I3C promoted senescence and inhibited proliferation, migration, and epithelial-mesenchymal transition in a wild-type UNC5B-dependent manner, primarily via SER604. Exogenous recombinant Netrin-1 could antagonize the aforementioned effects, whereas UNC5B knockdown abrogated this antagonistic action. In vivo, I3C suppressed tumor growth dependently on Netrin-1/UNC5B axis. CONCLUSIONS:I3C promotes CRC cellular senescence and suppresses tumor progression by blocking Netrin-1/UNC5B binding, likely through modulating Wnt/β-catenin pathway. This reveals a novel anticancer mechanism of I3C and a potential CRC therapeutic target.
Acute kidney injury (AKI) is a clinically significant syndrome characterized by rapid deterioration of renal function. Despite its complex and multifactorial pathogenesis, effective targeted therapies remain scarce. Mitochondrial dysfunction is increasingly recognized as a central driver of AKI progression. Mitochondrial transcription factor A (TFAM), a nucleus-encoded protein that governs mitochondrial DNA (mtDNA) maintenance, transcription and replication, plays an essential role in preserving mitochondrial integrity and biogenesis. This review systematically synthesizes current knowledge on TFAM biology, with a focus on its structural features, regulatory networks, and dynamic changes in the context of AKI. We integrate evidence showing that TFAM upregulation, whether through pharmacological interventions or genetic manipulation, consistently protects against tubular cell injury, preserves mitochondrial function, and attenuates inflammation across diverse AKI models. By providing a conceptual framework that links TFAM's molecular functions to its pathophysiological roles in the kidney, this review highlights TFAM as a promising therapeutic node. We also identify key knowledge gaps and propose future research directions to facilitate the translation of TFAM-targeted strategies into clinical practice.
Type I collagen is a major fibrillar component of the tumor extracellular matrix (ECM) and functions as a dynamic extracellular signaling platform. Its biological output is shaped by the type I collagen matrix state, comprising collagen abundance, trimeric composition, post-translational modifications, cross-linking, fibrillar organization and alignment, proteolytic turnover, cellular origin, and spatial distribution. This collagen-specific state is one component of the broader ECM state and regulates tissue mechanics, ligand accessibility, cell-matrix force transmission, vascular perfusion, and immune-cell positioning. In collagen-rich tumors, collagen-binding receptors, including integrins, discoidin domain receptors 1/2 (DDR1/DDR2), and leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), function as proximal sensors of collagen ligands. Together with Piezo1-mediated mechanosensing and mechanochemical signaling involving transforming growth factor-β (TGF-β)/SMAD and RhoA/ROCK-YAP/TAZ, these receptor systems translate collagen composition, architecture, and mechanics into adhesion, migration, epithelial-mesenchymal plasticity, mechanoadaptation, immune suppression, and therapeutic resistance. These responses vary with tumor type, disease stage, cancer-associated fibroblast states, immune composition, and organ microenvironment. This review integrates type I collagen matrix formation, maturation, state-dependent receptor engagement, mechanotransduction, tissue-level consequences, and therapeutic targeting. Multidimensional assessment of pathological type I collagen matrix states may guide matrix normalization and receptor-selective therapeutic strategies.
Renal cell carcinoma (RCC) is a highly aggressive malignancy often characterized by metabolic rewiring and limited long-term responses to targeted therapies. In this study, we unveil a previously uncharacterized tumor-suppressive paradigm driven by the m6A reader YTHDF1. Through multi-omics integration and clinical validation, we demonstrate that YTHDF1 is downregulated in RCC and serves as a clinically relevant biomarker associated with favorable clinicopathological features. Mechanistically, YTHDF1 functions in an m6A-dependent manner to directly promote the translational efficiency of the epigenetic modifier SETD2. SETD2 functions as a transcriptional co-activator for p53, triggering the transactivation of the p53/TIGAR metabolic checkpoint to block aerobic glycolysis. Disruption of the YTHDF1/SETD2/p53 axis induces enhanced glycolysis, which fuels tumor proliferation and metastasis. Importantly, we show that genetic overexpression of YTHDF1 not only restricts RCC aggressiveness but also enhances the therapeutic efficacy of sunitinib both in vitro and in vivo. Collectively, our findings identify the YTHDF1/SETD2/p53 axis as a critical translational-epigenetic-metabolic relay, providing a compelling rationale for leveraging epitranscriptomic machineries to improve therapeutic sensitivity in advanced renal cancer.
BACKGROUND:Osimertinib resistance is a major challenge in the treatment of EGFR-mutated lung adenocarcinoma (LUAD), and the role of LINC00511 in this process remains unclear. METHODS:We analyzed LINC00511 expression, patient prognosis, and its correlation with EIF2AK3 (encoding PERK) in LUAD using the GEPIA2 database, and further validated its circulating levels in patient peripheral blood by RT-qPCR. In vitro experiments, we constructed osimertinib-resistant H1650-OS and H1975-OS cell lines and regulated the expression of LINC00511 using genetic means. We evaluated the effects on resistance through functional experiments and used Western blot to detect PERK/Nrf2 and ER stress (ERS)-related proteins. Finally, we established a xenograft model to validate the in vitro findings in vivo. RESULTS:The results showed that elevated LINC00511 levels were observed in patients with osimertinib resistance, and these levels were positively correlated with PERK. Knockdown of LINC00511 could block the activation of the PERK/Nrf2 axis, inducing ERS. In vivo experiments confirmed that silencing LINC00511 enhances the inhibitory effect of osimertinib on osimertinib-resistant LUAD. Mechanistically, LINC00511 inhibits ERS through the PERK-Nrf2 pathway, thereby driving resistance. CONCLUSION:These findings provide clinical evidence for the early diagnosis and risk stratification of Osimertinib resistance, and suggest that targeting the LINC00511-PERK-Nrf2 axis may become a novel therapeutic strategy for restoring drug sensitivity, offering potential targets for the development of intelligent diagnostic and therapeutic nanomaterials.
Tumor necrosis factor receptor-associated factor 4 (TRAF4) is a RING-type E3 ubiquitin ligase linked to vascular inflammation, but whether it drives endothelial ferroptosis in atherosclerosis (AS) remains unclear. TRAF4 expression was assessed in GEO dataset GSE166780, ApoE-/- mice fed a high-fat diet for 4 months, and oxidized low-density lipoprotein (ox-LDL)-treated human coronary artery endothelial cells (HCAECs; 50 μg/mL, 24 h). In vivo, mice received tail-vein delivery of shTRAF4 vector. Plaque burden, endothelial injury, inflammatory responses, and ferroptosis were evaluated by histology, qRT-PCR, western blotting, immunofluorescence, reactive oxygen species (ROS) staining, and biochemical assays. TRAF4-PRMT5 and PRMT5-GPX4 interactions were examined by co-immunoprecipitation and GST pull-down. PRMT5 ubiquitination and protein stability were analyzed using MG132 and cycloheximide assays, and rescue experiments were performed by co-silencing TRAF4 and PRMT5. Our results showed that TRAF4 was upregulated in atherosclerotic tissues, the public dataset, and ox-LDL-treated HCAECs. TRAF4 knockdown reduced plaque burden, VCAM-1 expression, serum lipids and inflammatory cytokines, and attenuated ROS accumulation, ferrous iron (Fe2+) overload, lipid peroxidation, and Glutathione peroxidase 4 (GPX4) loss in vivo. In HCAECs, TRAF4 silencing improved viability, nitric oxide production, and mitochondrial function while reducing apoptosis, monocyte adhesion, inflammatory cytokine release, and ferroptosis. Mechanistically, TRAF4 directly bound PRMT5 and promoted its K48-linked ubiquitin-dependent degradation. CD31/TRAF4 co-staining confirmed endothelial TRAF4 enrichment in atherosclerotic tissue. PRMT5 bound GPX4, enhanced GPX4 methylation, and stabilized GPX4 protein. PRMT5 knockdown partially reversed the anti-ferroptotic effects of TRAF4 deficiency. Overall, TRAF4 promotes endothelial ferroptosis and AS progression by driving PRMT5 ubiquitination and destabilizing GPX4. The TRAF4-PRMT5-GPX4 axis may represent a therapeutic target in AS.
Neurokinin-1 (NK-1) is a G protein-coupled receptor for substance P and plays important roles in regulating diverse physiological and pathological processes. Previous studies have shown that NK-1 can modulate cell apoptosis in various malignancies, including triple-negative breast cancer (TNBC). This study aims to investigate the role of GTPase-activating protein (SH3 domain)-binding protein 2 (G3BP2) SUMOylation in NK-1-regulated progression in TNBC. Co-immunoprecipitation coupled with mass spectrometry was used to identify NK-1-interacting proteins and validate G3BP2, followed by truncation mapping, immunohistochemistry, functional assays, and site-directed mutagenesis to define its binding region, clinical relevance, and SUMOylation sites in TNBC. G3BP2 was identified as an NK-1-binding protein and was overexpressed in TNBC, correlating with poor overall survival. G3BP2 SUMOylation could enhance protein stability, thereby promoting TNBC cell proliferation and inhibiting apoptosis. G3BP2 interacted with NK-1 to facilitate UBC9/SUMO1-mediated SUMOylation at K281. Our findings demonstrate that NK-1 contributes to TNBC progression by regulating G3BP2 SUMOylation, revealing a critical post-translational modification mechanism underlying TNBC pathogenesis and highlighting the NK-1/G3BP2 SUMOylation axis as a potential therapeutic target.
Objective This study aimed to investigate how β-sitosterol (SIT) suppresses hypoxia-driven metastasis and angiogenesis in colorectal cancer. Methods We employed hypoxic colorectal cancer cells. The biological functions of SIT were assessed via in vitro assays for cell proliferation, migration, invasion, apoptosis and angiogenesis. A mouse model of colorectal cancer lung metastasis was constructed for grouped SIT intervention. Histopathology, immunofluorescence and Western blot were applied to detect the expression of key molecules. Results In vitro, SIT dose-dependently inhibited proliferation, migration and invasion of hypoxic colorectal cancer cells and triggered apoptosis. Additionally, SIT markedly inhibited angiogenesis and reversed the epithelial-mesenchymal transition (EMT). In the lung metastasis mouse model, SIT reduced metastatic foci, relieved lung injury, reversed EMT, decreased CD11b+Gr1+ myeloid cell infiltration, and suppressed FAP, eNOS and DDAH2 (angiogenesis-related factors). Further mechanistic studies revealed that SIT downregulated HIF-1α and VEGFA to mediate its anti-tumor actions. Conclusion Collectively, SIT blocked hypoxia-induced colorectal cancer metastasis and reversed the EMT through the HIF-1α/VEGF axis. Our findings offer new insights and potential therapeutic approaches for lung metastases from colorectal cancer.
FOXA1 and the androgen receptor (AR) are essential for prostate cancer cell survival. Although current targeted therapies against AR are widely used, they frequently lead to drug resistance, partly due to alterations in FOXA1. Minocycline, a commonly prescribed antibiotic, has demonstrated anticancer properties; however, its functional impact on FOXA1 and AR signaling remains unclear. In this study, we investigated whether minocycline suppresses tumor growth by targeting FOXA1 and its mutants while simultaneously inhibiting AR signaling, employing bioinformatics analyses, cellular assays (Western blot, qPCR, ChIP), and in vivo xenograft models. Our results show that minocycline binds directly to FOXA1 and markedly inhibits downstream oncogenic signaling mediated by the FOXA1-AR axis, thereby reducing proliferation and tumor growth. Importantly, in the presence of prevalent FOXA1 mutations (M253K and H247Y), minocycline retains potent inhibitory activity against metastatic prostate cancer driven by these FOXA1 mutants. Collectively, these findings identify minocycline as a novel agent that targets FOXA1 (including its mutants) and suppresses AR signaling, highlighting its potential for therapeutic repurposing and offering a clinically translatable strategy to overcome resistance in prostate cancer.
