Synovial Sarcoma (SS) is driven by the SS18::SSX fusion oncoprotein and is ultimately refractory to therapeutic approaches. SS18::SSX alters ATP-dependent chromatin remodeling BAF (mammalian SWI/SNF) complexes, leading to the degradation of canonical (cBAF) complexes and amplified expression of SS18::SSX-containing non-canonical BAF (ncBAF or GBAF) complexes that drive an SS-specific transcription program and tumorigenesis. We demonstrate that SS18::SSX activates the SUMOylation program. The small molecule SUMOylation inhibitor, TAK-981, de-SUMOylates the cBAF/PBAF component, SMARCE1, stabilizing and restoring cBAF on chromatin, shifting SS models away from SS18::SSX-driven transcription. The result is DNA damage, cell death and tumor inhibition across both human and mouse SS tumor models. TAK-981 synergizes with cytotoxic chemotherapy through increased DNA damage, leading to tumor regression. Targeting the SUMOylation pathway in SS restores cBAF complexes and blocks the SS18::SSX transcriptome, identifying an unappreciated role of SUMOylation in SS and a subsequent therapeutic vulnerability.

Targeted therapies in cancer are limited by cells exhibiting drug tolerance. We aimed to target drug tolerance in order to delay the development of acquired resistance. In melanoma, tolerance to MAPK pathway inhibitors is associated with loss of SOX10 and an enhanced TEAD transcriptional program. We show that loss of SOX10 is sufficient to up-regulate TEAD targets with dependence on the co-activator, TAZ. Active TAZ is sufficient to mediate tolerance to BRAF inhibitors and MEK inhibitors. We develop two covalent inhibitors, OPN-9643 and OPN-9652, designed to target the central palmitate binding pocket of TEADs. In SOX10-deficient cells, OPN-9643 and OPN-9652 reduce TEAD-dependent reporter activity and expression of TEAD targets, CTGF and CYR61. OPN-9643 and OPN-9652 treatment enhances the inhibitory effects of MAPK-targeted therapies in 2D and 3D growth assays in SOX10 knockout cells and reverses tolerance mediated by active TAZ. In vivo, OPN-9652 delays the onset of acquired resistance to BRAF inhibitors and MEK inhibitors from minimal residual disease. Thus, TAZ-TEAD activity plays an important role in melanoma drug tolerance and the development of acquired resistance.

Grapevine downy mildew, caused by Plasmopara viticola, poses significant threats to viticulture, necessitating sustainable alternatives to chemical fungicides. This study investigates the efficacy of exogenous 24-epibrassinolide (EBR), a brassinosteroid, in enhancing disease resistance across two grape cultivars: susceptible Vitis vinifera Cabernet Sauvignon and resistant Ecolly. EBR application reduced P. viticola colonization in both cultivars, suppressing hyphal growth and sporulation. In Cabernet Sauvignon, EBR delayed pathogen establishment by modulating stomatal closure and boosting antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase, peroxidase) and pathogenesis-related proteins (chitinase, β-1,3-glucanase). Ecolly exhibited inherent resistance, characterized by localized hypersensitive responses and elevated salicylic acid (SA) and jasmonic acid (JA) levels post-EBR treatment, which amplified systemic acquired resistance (SAR) via upregulation of PAD4, EDS1, and PRs genes. Stomatal regulation in Cabernet Sauvignon correlated with EBR-induced abscisic acid (ABA) dynamics, while Ecolly maintained constitutive stomatal defense. Transcriptional analysis revealed cultivar-specific phytohormonal interplay, with SA pathways dominating in Ecolly and delayed SA accumulation in Cabernet Sauvignon. These findings demonstrate that EBR enhances resistance through multifaceted mechanisms-pathogen inhibition, redox homeostasis, hormonal signaling, and genetic priming-with efficacy modulated by genetic background. This study underscores brassinosteroid as a sustainable tool for integrated disease management in grapevines.

This study first demonstrates that PsGSH2 enhances cadmium tolerance not only by boosting antioxidant defense but also by modulating metal transporter genes to reduce Cd accumulation in plants. Cadmium (Cd) stress poses a significant environmental issue. Potentilla sericea, characterized by strong resistance, is an excellent groundcover for pollution remediation. Glutathione synthetase is one of the key enzymes that promote the synthesis of the antioxidant glutathione (GSH). We cloned PsGSH2, which was up-regulated under Cd stress, and introduced it into Arabidopsis thaliana to validate the response of transgenic lines to Cd. The results showed that the expression level of PsGSH2 was significantly up-regulated (15.45-fold) in the roots of P. sericea under cadmium stress. Overexpression (OE) of PsGSH2 in A. thaliana significantly enhanced Cd tolerance. Compared to wild-type (WT) plants, OE lines exhibited a more than sevenfold increase in seed germination rate under Cd stress, with a significantly reduced biomass loss (< 40%). The transgenic lines showed enhanced photosynthetic performance, a reinforced antioxidant system (up to 1.9- and 2.2-fold higher than WT), and reduced oxidative damage (50-75% of WT). Crucially, they exhibited a 59.05% reduction in shoot Cd accumulation, supported by significantly lower bioconcentration factor and transport factor values (46.12% and 69.17%, respectively). Molecular analysis revealed upregulation (1.84- to 5.44-fold) of key genes related to Cd detoxification (AtGSH1, AtGSH2, AtIRT1, AtPCR1, AtPCR2, AtMT3). Therefore, this study provides valuable insights for developing Cd-tolerant plants through genetic engineering approaches, laying the foundation for further research on Cd resistance in P. sericea.

This narrative review examines recent advances in understanding the epigenetic and transcriptional dynamics of the central-medial amygdala across different stages of ethanol use. It covers the various models and protocols of ethanol administration, emphasizing their strengths and limitations in helping understand ethanol-induced changes in brain and behaviour. The findings from protocols utilizing acute, chronic-intermittent ethanol (CIE) and chronic-continuous ethanol (CCE) exposure are summarized into a proposed mechanism of epigenetic dysregulation and neuroadaptation, spanning from initial ethanol exposure to withdrawal and its influence on gene expression and neuronal activity. This review also explores potential epigenetic targets for therapeutic intervention. Understanding the mechanisms that underlie histone remodelling during initial ethanol exposure, chronic exposure, and withdrawal has the potential to elucidate novel cessation strategies tailored to specific stages of alcohol use disorder (AUD).

Multidrug resistance (MDR) is a major challenge in cancer chemotherapy. A critical factor contributing to MDR is overexpression of ATP-binding cassette (ABC) transporters, such as ABCB1. Novel alternative therapeutic strategies are needed to overcome resistance associated with ABC transporters. In the present study, we aimed to elucidate the mechanisms by which melatonin overcomes ABCB1-mediated MDR in cancer cells, with a focus on mitochondrial function. We analyzed the effects of melatonin (1 mM) on head and neck squamous cell carcinoma cell lines (CAL 27 and SCC-9) overexpressing ABCB1 and exhibiting increased resistance to cisplatin (CDDP) compared to their parental cells. To further validate the role of melatonin in reversing ABCB1-mediated MDR, we also evaluated its effects on doxorubicin-resistant MCF-7 breast cancer cells. We further examined the potential of melatonin to overcome MDR in CAL 27 xenografted mice. Here, we report that melatonin treatment specifically triggered reactive oxygen species (ROS) production in mitochondria and weakened chemoresistance. ROS oxidized NADH into NAD+, and limiting the availability of ATP for efflux pump activity. Additionally, melatonin decreased the number of mitochondria localized near the nucleus instead of the cytoplasm and downregulated ABCB1 expression. Intratumoral administration of melatonin effectively overcame CDDP resistance in CAL 27/ABCB1 xenografts, significantly reducing tumor volume and promoting apoptosis. These findings demonstrate that melatonin enhances chemosensitivity in ABCB1-overexpressing cells by modulating mitochondrial metabolism, redox balance, and ABCB1 expression, highlighting its potential as an adjuvant therapy to overcome MDR.

Gastric cancer (GC) is a common type of cancer known for its challenges in early detection and unfavorable prognosis. The pathway involving Wnt/β-catenin and the improper regulation of microRNAs (miRNAs), especially miR-27a-3p, are crucial in the advancement of GC. Ursolic acid (UA), which is a naturally occurring anticancer agent, shows promise in the inhibition of GC. Although UA's anticancer effects have been recognized, the underlying molecular mechanisms in GC remain incompletely defined. Our findings indicate that UA strongly restricts the expansion, motility, and invasive behavior of GC cells by dampening activity within the Wnt/β-catenin cascade. Treatment with UA lowered miR-27a-3p expression, and blocking this miRNA further curtailed tumor cell aggressiveness by restoring DKK2, which functions as a suppressor of Wnt-driven signaling. The protein under investigation showed lower expression in advanced tumors. Its expression in these advanced tumors correlated with better pathologic outcomes and survival prognosis. Thus, we can categorize this protein as a novel tumor suppressor in GC. Consistent with these in vitro results, in vivo assays demonstrated that UA effectively curtailed tumor development. Taken together, these findings indicate that UA restricts GC progression via modulation of the miR-27a-3p/DKK2/Wnt/β-catenin axis, providing mechanistic insights for potential therapeutic strategies.

Substance use disorders, particularly drug addiction, are complex neurophysiological conditions characterized by cycles of compulsive drug use, withdrawal symptoms, and relapses. Methamphetamine (MA) addiction evolves through repeated exposure, altering brain circuits related to reward and neuroplasticity. The need for reliable biomarkers to diagnose and monitor MA addiction has become increasingly critical in clinical practice. In this study, we explored the time-dependent transcriptomic changes in the rat striatum immediately after short-term abstinence following MA self-administration. Using a rat model, we conducted RNA sequencing to analyze the transcriptomic alterations in the striatum immediately after the self-administration and short-term abstinence phases (12- and 24-h post-MA). Through protein-protein interaction (PPI) network analysis and gene expression pattern assessment, we identified key genes that demonstrated significant expression changes. These genes were strongly linked to reward mechanisms, synaptic plasticity, and memory processes, suggesting a role in mediating MA-associated behaviors. Understanding the expression dynamics of these genes provides valuable insights into the molecular mechanisms underlying MA addiction and offers a foundation for developing diagnostic tools and therapeutic strategies targeting addiction-related neural adaptations.

Drug synergy in cancer therapy has gained attention for its potential to enhance efficacy and minimize adverse effects. Synergy, where the combined effect of drugs exceeds the sum of their individual effects, is beneficial in addressing tumor heterogeneity and drug resistance in oncology. Esophageal squamous cell carcinoma (eSCC), accounting for over 85% of esophageal cancer cases, has a poor prognosis and limited therapeutic options. Targeted therapies have shown inefficiency in eSCC due to resistance mechanisms, making drug combination an interesting option.

Multiple myeloma (MM) is a neoplasm of antibody-producing plasma cells and is the second most prevalent hematological malignancy worldwide. Development of drug resistance and disease relapse significantly impede the success of MM treatment, highlighting the critical need to discover novel therapeutic targets. In a custom CRISPR/Cas9 screen targeting 197 DNA damage response-related genes, Protein Arginine N-Methyltransferase 1 (PRMT1) emerged as a top hit, revealing it as a potential therapeutic vulnerability and survival dependency in MM cells. PRMT1, a major Type I PRMT enzyme, catalyzes the asymmetric transfer of methyl groups to arginine residues, influencing gene transcription and protein function through post-translational modification. Dysregulation or overexpression of PRMT1 has been observed in various malignancies including MM and is linked to chemoresistance. Treatment with the Type I PRMT inhibitor GSK3368715 resulted in a dose-dependent reduction in cell survival across a panel of MM cell lines. This was accompanied by reduced levels of asymmetric dimethylation of arginine (ADMA) and increased arginine monomethylation (MMA) in MM cells. Cell cycle analysis revealed an accumulation of cells in the G0/G1 phase and a reduction in the S phase upon GSK3368715 treatment. Additionally, PRMT1 inhibition led to a significant downregulation of genes involved in cell proliferation, DNA replication, and DNA damage response (DDR), likely inducing genomic instability and impairing tumor growth. This was supported by Reverse Phase Protein Array (RPPA) analyses, which revealed a significant reduction in levels of proteins associated with cell cycle regulation and DDR pathways. Overall, our findings indicate that MM cells critically depend on PRMT1 for survival, highlighting the therapeutic potential of PRMT1 inhibition in treating MM.

Lysine (K)-specific demethylase (KDM) 6A, a histone modifier with inhibitory roles in many types of cancers, is frequently mutated in clear cell renal cell carcinoma (ccRCC). Here, we investigated the role of KDM6A in ccRCC progression, including its effect on resistance to tyrosine kinase inhibitors (TKIs). The clinical impact of KDM6A expression was examined by immunohistochemical analysis of nephrectomized tissues from patients with ccRCC. Upon generation of KDM6A-deficient RCC cells by CRISPR/Cas9-mediated gene editing, in vitro cancer cell property analysis, RNA sequencing, and in vivo murine xenograft model examination were performed. The relationship between KDM6A expression and efficacy of TKIs was analyzed using data from public databases and in vitro pharmacological assessments. Patients with KDM6A-low ccRCC had poor prognoses. Promoted invasion, migration, cancer stemness, epithelial-mesenchymal transition (EMT), and in vivo tumor progression were observed in KDM6A-deficient RCC cells. A reanalysis of previous clinical trial data revealed lower efficacy of sunitinib in patients with KDM6A-low ccRCC. Consistently, KDM6A-deficient cells showed resistance to cabozantinib and decreased expression of target molecules of TKIs. KDM6A deficiency contributes to ccRCC progression and TKI resistance, suggesting that targeting KDM6A-related pathways may offer new therapeutic strategies for patients with KDM6A-deficient ccRCC.

The impacts of oregano essential oil and sodium butyrate on the growth performance, hematological, biochemical, immunological, antioxidative, and gene expression profiles of Nile tilapia (Oreochromis niloticus) fingerlings were examined in this study. One hundred eighty healthy fingerlings (16.00 ± 2.00 g) were acclimated and accidentally divided into three groups, each further split into three replicates. The groups were fed a basal diet (control) or diets supplemented with 0.5% and 1% oregano essential oil and sodium butyrate for eight weeks (The feed additives were administered continuously throughout an 8-week trial period, simulating a practical feeding strategy that would be feasible during high-risk periods (e.g., post-handling stress, seasonal disease outbreaks). Fish fed the supplemented diets demonstrated significantly greater growth performance, with the oregano 1% + sodium butyrate 1% group achieving the highest final body weight (36.91 ± 0.66 g) and lowest feed conversion ratio (1.32 ± 0.04). Hematological indices, including RBC count, Hb, and PCV, were substantially improved. Biochemical analysis revealed reduced ALT, AST, urea, and creatinine levels alongside elevated total protein and globulin concentrations in treated groups. Immunological and antioxidative parameters, such as WBC count, phagocytic activity, SOD, and GPX, showed significant enhancements, particularly in the oregano 1% + sodium butyrate 1% group. Gene expression analysis exposed upregulation of immune-related (TNF-α, IL-1β) and antioxidative (SOD, GPX) genes in supplemented groups, highlighting their role in enhancing health and stress responses. Post Aeromonas hydrophila infection, the supplemented groups exhibited improved immunity, antioxidative status, and survival. These findings suggest that dietary supplementation with oregano essential oil and sodium butyrate, particularly at 1% inclusion, effectively enhances growth, health status, immunity, and gene expression in Nile tilapia.

Background Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related death worldwide. Lenvatinib is a common first-line treatment for advanced HCC. However, resistance to lenvatinib is the greatest challenge limiting its clinical application. Currently, the molecular mechanisms of resistance remain poorly understood. Methods The expression of DDX1 and Ephrin-A3 in lenvatinib-resistant HCC cells was identified via RNA-seq and Western blotting. Bioinformatic analyses were applied to explore its expression and prognostic role. The biological role of DDX1 was evaluated via CCK8, EdU, flow cytometry analyses and xenograft tumor model. The regulation between DDX1 and Ephrin-A3 was determined by mass spectrometry, coimmunoprecipitation, RNA Immunoprecipitation, and RNA stability assay. Results We successfully established lenvatinib-resistant HCC cells. The results of RNA-seq showed DDX1 and Ephrin-A3 were significantly increased in lenvatinib-resistant HCC cells compared to parental cell. The DDX1 expression in HCC tissues is positively associated with worse prognosis. DDX1 knockdown increased the sensitivity of cells to lenvatinib by inhibiting proliferation and promoting apoptosis in vitro and in vivo. Conversely, overexpression of DDX1 exhibited the opposite regulation. Moreover, DDX1 bound to Ephrin-A3 and regulated its expression levels. The effects of DDX1 overexpression on cell proliferation, apoptosis, and lenvatinib resistance were significantly blocked by Ephrin-A3 knockdown. Mechanistically, DDX1 promotes lenvatinib resistance in HCC by regulating Ephrin-A3 mRNA stability and activating the Wnt/β-catenin pathway. Conclusion: The increased DDX1 expression in HCC cells promotes lenvatinib resistance via regulating Ephrin-A3 mRNA stability and activating the Wnt/β-catenin pathway, indicating that targeting DDX1 may be an important strategy for overcoming lenvatinib resistance.

Renal ischemia-reperfusion injury (RIRI) is a major cause of acute kidney injury, with endothelial dysfunction playing a central role in its pathophysiology. However, the molecular mechanisms underlying endothelial damage during RIRI remain incompletely understood.

The widespread use of pesticides such as deltamethrin (a pyrethroid) and acetamiprid (a neonicotinoid) has sparked concerns regarding their effects on human health, particularly their potential role in carcinogenesis. This study investigated the cytotoxic, molecular, and functional effects of these pesticides, individually and in combination, on the MDA-MB-231 triple-negative breast cancer (TNBC) cell line. This model was chosen to specifically investigate estrogen recpetor (ER)-independent mechanisms due to its expression of targets such as aryl hydrocarbon receptor (AhR), peroxisome proliferator-activated receptor gamma (PPARγ), and G protein-coupled estrogen receptor (GPER); however, it does not reflect normal mammary cell responses. Cytotoxicity was assessed via XTT assays, migration was analyzed using wound-healing assays, and gene expression changes in AhR, PPARγ, and Caspase-3 were measured using RT-qPCR. Molecular docking was performed to predict pesticide-protein interactions, and in silico toxicity assessments using ProTox-II supplemented the in vitro results by predicting toxicity profiles relevant to public health. Both pesticides exhibited dose-dependent cytotoxicity, and their combination produced an additive effect on cell viability. Importantly, suppression of cell migration and downregulation of AhR and PPARγ expression reflected toxic stress responses at high pesticide concentrations, rather than therapeutic or anti-cancer potential. While apoptosis-related gene expression (Caspase-3) was increased, this effect did not reach statistical significance. Molecular docking supported strong interactions with key pathways related to xenobiotic metabolism and apoptosis. These findings emphasize that, at high and non-environmentally relevant concentrations, deltamethrin and acetamiprid induce additive cytotoxic effects and disrupt molecular processes in a mechanistic cancer model. The results highlight the need for further investigation using normal cell systems and environmentally relevant exposures to clarify real-world risk and biological mechanisms, and should not be interpreted as evidence of therapeutic activity. This study underscores the mechanistic relevance of pesticide exposure in environmental toxicology rather than any potential therapeutic application.

Costunolide (COS) is a bioactive sesquiterpene lactone compound extracted from Aucklandia lappa, known for its anticancer, anti-inflammatory, and antioxidant properties, aligning with the traditional Chinese medicine theory of "clearing heat and dispersing nodules" in the treatment of breast ailments. Network pharmacology identified EGFR as the core target of COS, with enrichment analysis revealing the EGFR/ERK/AKT axis as a key pathway. Molecular docking demonstrated strong binding affinity of COS to the EGFR kinase domain, relying on hydrogen bonds and hydrophobic interactions. In vitro experiments showed that COS inhibited TNBC cell proliferation and induced apoptosis. Mechanistically, COS increased EGFR ubiquitination, leading to a decrease in EGFR protein levels, thereby inhibiting EGFR phosphorylation and the activation of downstream ERK and Akt signaling pathways. EGF could partially reverse the growth inhibitory effects of COS, confirming the critical role of EGFR. This study elucidates that COS exerts its anti-TNBC effects by inducing EGFR ubiquitination and degradation, thereby inhibiting the ERK/AKT signaling pathway. This finding integrates traditional Chinese medicine theory with modern molecular oncology mechanisms, providing a reference for the development of plant-derived multi-target anticancer drugs.

This study evaluated the physiological, biochemical traits, molecular responses, and vase life of cut Alstroemeria hybrida 'Amatista' in response to preharvest applications of putrescine and melatonin. A factorial experiment was arranged in a completely randomized design with three replications. The treatments included foliar sprays of putrescine (0, 1.5, and 3 mM), melatonin (0, 50, and 100 µM), and four postharvest sampling times (0, 5, 10, and 15 days). Measured parameters included chlorophyll index, relative fresh weight, solution uptake, ion leakage, malondialdehyde (MDA), hydrogen peroxide (H2O2), polyphenol oxidase (PPO) activity, vase life, and the relative expression of chlorophyllase (CHL) and ACC oxidase (ACO) genes. Postharvest senescence was associated with a time-dependent decline in chlorophyll index, water content, and solution uptake, along with increased ion leakage, MDA, H₂O₂ levels, and PPO activity. However, preharvest application of putrescine and melatonin effectively delayed these detrimental changes. Notably, the combined application of 1.5 mM putrescine and 50 µM melatonin resulted in the greatest improvement in postharvest performance, extending vase life from 16.00 to 23.66 days and enhancing physiological and biochemical traits. Moreover, gene expression analysis indicated that these treatments suppressed the expression of CHL and ACO, suggesting a molecular mechanism involved in delayed senescence. Overall, the findings highlight the potential of putrescine and melatonin as eco-friendly preharvest treatments to improve the postharvest quality and longevity of cut Alstroemeria flowers by modulating key physiological, biochemical, and molecular processes.

Due to a high rate of recurrence coupled with resistance towards modern therapies, colorectal cancer (CRC) is considered as the third cause of cancer-related death worldwide. Gemini curcumin (Gemini-Cur) is one of the last nanoformulation of curcumin with significant toxicity on colorectal cancer. Herein, we aimed to unravel the modulated lncRNAs, related mRNAs and downstream cellular pathways in Gemini-Cur treated HT-29 colorectal cancer cells. 9805 lncRNAs were found to be differentially expressed in nanocurcumin-treated cancer cells versus non treated group. The top 20 lncRNAs were selected for further studies and, 14,472 co-expression relationships between these RNAs and 70,711 mRNAs were identified. Among top 20 lncRNAs, tumor-suppressive C8orf31 and ARHGAP5-AS1, as well as oncogenic XIST, FTX, and NEAT1 were the most notable due to their involvements in cancer-related cellular pathways. Functional enrichment analyses demonstrated that the modulated lncRNAs and their targets are involved in cell cycle, p53 signaling, translation, and helicase activity pathways. In conclusion, our study elucidated new molecular mechanisms of nano-curcumin in the regulation of lncRNA expression and the discovery of potential targets in therapeutic interventions for CRC. More studies are needed to confirm the therapeutic implications of these findings.

Dapagliflozin (DAPA), an SGLT-2 inhibitor, shows peritoneal protection and can alleviate high glucose-induced peritoneal fibrosis. Yet, its precise molecular mechanism is unknown. This study aims to explore DAPA's protective effect on the peritoneum and its underlying mechanism. In vitro, human peritoneal mesothelial cells (HPMCs) were isolated from peritoneal dialysate and cultured. HMrSV5 cells were stimulated with 2.5% D-Glucose (high glucose, HG) for 48 h, then cultured in D-glucose DMEM medium with or without DAPA. To assess SGLT2i-induced ENKUR down-regulation, HMrSV5 cells were treated with DAPA for 24 h while overexpressing ENKUR. In vivo, six-week-old male Sprague-Dawley rats were treated with high-glucose dialysate via an intraperitoneal catheter, with or without addition of DAPA. Changes in SGLT2, ENKUR, PI3K/AKT pathways, and EMT markers were evaluated in HPMCs and the rat model. As dialysis duration increases the morphology of the cells transitioned from a cobblestone appearance to a spindle shape. Immunofluorescence analysis confirmed the mesothelial cell origin and revealed an upregulation of ENKUR and the PI3K/AKT signaling pathway, which are associated with the occurrence of EMT. DAPA was found to decrease the expression of ENKUR and inhibit the activation of the PI3K/AKT pathway induced by high glucose in HMrSV5 cells. In rats subjected to PD, we observed a reduction in ultrafiltration capacity, an increase in peritoneal thickness, and elevated levels of SGLT2, ENKUR, PI3K/AKT and EMT markers. Notably, these alterations were mitigated by intragastric administration of DAPA. DAPA effectively ameliorates high glucose-induced peritoneal fibrosis through downregulation of ENKUR/PI3K/AKT signaling pathway.

Ferroptosis is an iron-dependent cell death pathway that involves multiple genes, including the transferrin receptor (TFRC/CD71), glutathione peroxidase 4 (GPX4) and cystine-glutamate antiporter (SLC7A11). This study is based on the hypothesis that orphan nuclear receptor 4A1 (NR4A1) and NR4A2 maintain low levels of ferroptosis in triple negative breast cancer (TNBC) cells and bis-indole derived (CDIM) compounds act as NR4A1/2 ligands that induce ferroptosis by enhancing CD71 expression. 1,1-Bis(3'-indolyl)-1-(3,5-disubstitutedphenyl)methane (DIM-3,5) analogs were investigated for their cytotoxicity and effects on NR4A1 and NR4A2 regulated genes and induction of ferroptosis. Several assays also determined enhanced lipoperoxidation, reactive oxygen species and malondialdehyde formation in TNBC cells. Knockdown of NR4A1, NR4A2, Sp1 and Sp4 was carried out by RNA interference. Molecular mechanisms of NR4A1/2-mediated regulation of CD71 expression were determined using CD71-luciferase promoter constructs, overexpression of Sp1 and chromatin immunoprecipitation (ChIP) assays. Initial studies show that DIM-3,5 analogs act as an inverse NR4A1/NR4A2 agonists that downregulate the pro-oncogenic responses/gene products regulated by both receptors in TNBC cells. DIM-3,5 analogs also induced ROS, malondialdehyde and lipoperoxide formation in TNBC cells, and this was accompanied by decreased expression of GPX4 and SLC7A11 and induction of CD71. Induction of CD71, an important biomarker of ferroptosis was observed after treatment of TNBC cells with DIM-3,5 analogs, knockdown of NR4A1, NR4A2, Sp1 or Sp4 demonstrating that induction of CD71 was coregulated by both receptors. Moreover, both promoter and ChIP analysis indicated that NR4A1 and NR4A2 acted as ligand-dependent cofactors of Sp1/4-mediated expression of CD71 in TNBC cells. Thus, CD71, a key biomarker of ferroptosis is an NR4A1/2/Sp regulated gene that can be directly targeted by DIM-3,5 inverse NR4A1/2 agonists to induce ferroptosis in TNBC cells.