Background: This study systematically investigates the therapeutic effects of naringenin (NAR) and oleuropein (OLE) on prostate cancer through miR-155-5p regulation. Methods: Experimental studies conducted on MAT-LyLu prostate cancer cell lines revealed that the application of NAR (50 μM) and OLE (75 μM) significantly increased miR-155-5p expression by 2.89-fold and 1.74-fold, respectively (p < 0.05). Bioinformatics analyses have indicated that miR-155-5p interacts with critical oncogenic pathways such as KRAS, CDK2, NF-κB, and TGF-β/Smad2. Computational analyses have revealed that miR-155-5p interacts with 16 critical oncogenic targets, including KRAS and CDK2. Molecular docking studies showed that NAR binds to the Switch I/II region of KRAS with a binding energy of -8.2 kcal/mol, while OLE binds to the ATP-binding pocket of CDK2 with an affinity of -9.1 kcal/mol. Pharmacokinetic evaluations revealed that NAR indicated high oral bioavailability (93.763% HIA) and full compliance with Lipinski's rules, while OLE required advanced formulation strategies due to its high polarity. Network pharmacology analyses have shown that NAR affects lysosomal functions and enzyme regulation, while OLE affects G protein-coupled receptors and oxidoreductase activity. Results: Results indicate that NAR and OLE exhibit antitumor effects through multiple mechanisms by increasing miR-155-5p expression and inhibiting critical oncogenic targets in prostate cancer. Conclusions: Findings suggest that the dietary intake of these natural compounds (citrus and olive products) should be considered in prostate cancer prevention strategies, shedding light on the epigenetic mechanisms of polyphenols in cancer treatment and contributing to the development of new therapeutic strategies.

Recurrent endometrial cancer (EC) has limited therapeutic options beyond platinum-based chemotherapy, highlighting the need to identify exploitable molecular vulnerabilities. Tumors with high genomic instability, including microsatellite instability-high (MSI-h) or copy-number-high (CNH) ECs, rely on the ATR-CHK1 signaling pathway to tolerate replication stress and maintain genome integrity, making this pathway an attractive therapeutic target. However, acquired resistance to ATR and CHK1 inhibitors (ATRi/CHK1i) often develops, and the transcriptomic basis of this resistance in EC remains unknown. Here, we established isogenic ATRi- and CHK1i-resistant cell line models from MSI-h (HEC1A) and CNH (ARK2) EC lineages and performed baseline transcriptomic profiling to characterize stable resistance-associated states. MSI-h-derived resistant clones adopted a unified transcriptional state enriched for epithelial-mesenchymal transition, cytokine signaling, and interferon responses, while ATRi-resistant models showing additional enrichment of developmental and KRAS/Notch-associated pathways. In contrast, CNH-derived resistant clones diverged by inhibitor class, with ATRi resistance preferentially enriching proliferation-associated pathways and CHK1i resistance inducing interferon signaling. Notably, THBS1, EDN1, and TENM2 were consistently upregulated across all resistant models relative to parental lines. Together, these findings demonstrate that acquired resistance to ATRi and CHK1i in EC is shaped by both lineage and inhibitor class and provide a transcriptomic framework that may inform future biomarker development and therapeutic strategies.

Neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD) are characterized by complex pathologies with progressive neurodegeneration, protein misfolding, oxidative stress, and persistent inflammation. Recent findings indicate the pivotal involvement of epigenetic disruption, particularly aberrant histone deacetylase (HDAC) activity, in disease initiation and progression. In the current review, we systematically discuss the mechanistic function of HDACs across all classes (I, IIa, IIb, III, and IV) in neurodegenerative disease mechanisms, such as their involvement in the modulation of gene expression, mitochondrial function, proteostasis, and neuronal survival. We discuss the therapeutic potential, as well as limitations, of HDAC inhibitors (HDACis), such as pan-inhibitors and isoenzyme-selective inhibitors, and new multi-target-directed ligands with HDAC inhibition combined with acetylcholinesterase modulation, PDE modulation, MAO-B inhibition, or NMDAR modulation. Particular emphasis is placed on the development of HDAC6-selective inhibitors with enhanced brain permeability and reduced toxicity, which have shown promising preclinical efficacy in ameliorating hallmark pathologies of AD, PD, and HD. In addition, s-triazine-based scaffolds have recently emerged as promising chemotypes in HDAC inhibitor design, offering favorable pharmacokinetic profiles, metabolic stability, and the potential for dual-target modulation relevant to neurodegeneration. The review also explores the future of HDAC-targeted therapies, including PROTAC degraders, dual-inhibitor scaffolds, and sustainable, BBB-penetrant molecules. Collectively, this review underscores the importance of HDAC modulation as a multifaceted strategy in the treatment of neurodegenerative diseases and highlights the need for continued innovation in epigenetic drug design.

Zinc (Zn) is a mineral micronutrient that is essential for plant growth and development. Soil Zn deficiency or excess severely impacts plant health and crop yields. MicroRNAs (miRNAs) play crucial roles in plant responses to abiotic stress, but their roles in Zn homeostasis in important crop bread wheat (Triticum aestivum L.) remain unknown. This study investigated miRNA expression profiles in wheat roots under different Zn supply conditions using high-throughput sequencing. Phenotypic and physiological analyses revealed that high Zn promoted wheat plant growth, while low and excess Zn resulted in wheat plant growth inhibition and oxidative stress. A total of 798 miRNAs (including 70 known and 728 novel miRNAs) were identified; among them, 10 known and 122 novel miRNAs were differentially expressed. Many key miRNAs, such as miR397-5p, miR398, 4D_25791, and 5A_27668, are up-regulated under low Zn but down-regulated under high Zn and excess Zn. Target gene prediction and enrichment analysis revealed that the regulated genes of these miRNAs focused on "zinc ion transmembrane transporter activity", "divalent inorganic cation transmembrane transporter activity", and "cellular detoxification" processes in the low Zn vs. CK group. However, "glutathione metabolism" and "ABC transporter" pathways were obviously enriched in high Zn vs. excess Zn conditions, implying their potential functions in alleviating the oxidative damage and Zn efflux caused by Zn toxicity. Together, this study identified key miRNAs that respond to both Zn deficiency and excess Zn in bread wheat, revealing distinct regulatory patterns of the target genes in different Zn supply conditions. These findings provide a new field and valuable candidate miRNAs for molecular breeding aimed at improving zinc's utilization efficiency in wheat.

Flavonols are an important secondary metabolite in grape, which play a crucial role in plant growth and development, human health, and wine making. Ethylene and its inhibitor 1-Methylcyclopropene (1-MCP) are widely used in grape berry production. However, the regulation mechanism of flavonol biosynthesis by ethylene and 1-MCP remains elusive in yellow-green grape varieties. Here, the content of flavonols in 'Chardonnay' grape berry skin after ethylene treatment was significantly higher than the control, while 1-MCP treatment was lower than the control. The phenylpropanoid biosynthesis-related genes and a transcription factor VvERF003 were screened for possible involvement in ethylene-mediated flavonol biosynthesis by transcriptome sequencing. The role of VvERF003 was further proved to promote flavonol accumulation in the transient overexpression of grape fruits and leaves, and the upregulation of genes related to flavonol biosynthesis. Furthermore, VvERF003 promoted flavonol biosynthesis by directly binding to and activating the promoters of VvCHI1 and VvFLS1, and positively regulated their expression. These results indicated that VvERF003 was induced by ethylene and promoted the accumulation of flavonols in 'Chardonnay' grape berry skin by positively regulating the flavonol biosynthesis genes VvCHI1 and VvFLS1.

Histone deacetylases (HDACs) are central epigenetic regulators in non-small cell lung cancer (NSCLC), yet responses to HDAC inhibitors (HDACi) vary markedly between lung adenocarcinoma (LUAD) and lung squamous carcinoma (LUSC). We asked how the pan-HDAC inhibitor suberoylanilide hydroxamic acid (SAHA, vorinostat) rewires lineage-specific transcriptional programs and whether SAHA-aligned modules of genes, rather than individual loci, capture clinically relevant vulnerabilities in each subtype.

Corneal transparency maintenance relies on the water-pumping function of the corneal endothelium. Currently, corneal transplantation remains the only available treatment for corneal endothelial dysfunction, therefore, the development of alternative therapies is critical due to the global shortage of donor corneas. In our previous study, we confirmed that corneal stromal cells (CSCs) secretion can promote corneal endothelial cells (CEnCs) proliferation. This effect can be enhanced by treatment with lysophosphatidic acid (LPA), a bioactive phospholipid. Nevertheless, the components involved in CSC secretion remain to be elucidated. In this study, we investigated the therapeutic potential of CSC-derived exosomes and exosomal microRNAs (miRNAs) for enhancing CEnCs proliferation and corneal endothelial healing. CSC exosomes were characterized via nanoparticle tracking (NTA), transmission electron microscopy (TEM), and immunoassays. The miRNA expression profiles of CSC exosomes were identified via RNA sequencing, revealing a total of 767 distinct miRNAs. The proliferative effects of CSC exosomes and exosomal miR-221-3p were increased by LPA. Ectopic expression of miR-221-3p further increased CEnC proliferation and suppressed the expression of the CDK inhibitor p27Kip1. The therapeutic efficacy was evaluated using a transcorneal freezing rabbit model, where intrastromal injection of CSC exosomes or ectopic expression of miR-221-3p significantly ameliorated corneal endothelial damage, as supported by improved in vivo corneal recovery, including restoration of corneal thickness, and re-establishment of a hexagonal morphology in the corneal endothelium. Our findings suggest that CSC exosomes and miR-221-3p represent potentially promising cell-free therapies for treating corneal endothelial diseases, highlighting an innovative approach to improving corneal regeneration.

Bacteria frequently encounter sub-inhibitory concentrations of antibiotics within host tissues and natural environments, where these exposures can function as regulatory signals rather than growth inhibitors. Streptococcus pyogenes, an important human pathogen, transitions between asymptomatic colonization and invasive disease through complex regulation of virulence factors and stress responses. In this article we aim to show that there is alteration in virulence of S. pyogenes on exposure to CWA antibiotics. In clinical practice, cell wall-active antibiotics remain the cornerstone of therapy against S. pyogenes, therefore understanding their ability to modulate virulence and stress responses is critical.

Gastric cancer represents the fifth most common malignancy globally and the third leading cause of cancer-related deaths. Due to its insidious early symptoms and frequent metastasis at diagnosis, the survival rate remains dismal. There is thus an urgent clinical need for novel therapeutic agents. Innovative strategies combining traditional chemotherapy with interventions that induce novel cell death pathways represent a promising translational direction for improving patient outcomes. Punicalagin (PUN), a natural polyphenol derived from pomegranate, exhibits potent antioxidant and broad-spectrum antitumor activities, yet its role in gastric cancer remains understudied. Three gastric cancer cell lines (AGS, HGC27, MFC) and one normal gastric mucosal epithelial cell line (GES-1) were initially selected for in vitro experiments. The effects of PUN on gastric cancer cells and normal gastric mucosal epithelial cells were assessed through MTT assay, propidium iodide (PI) staining, and cell colony formation assays, while cell migration ability was evaluated using a scratch wound healing assay. The inhibitory effect of PUN on gastric cancer was tested in a subcutaneous tumor model in nude mice, with pathological changes in vital tissues and organs observed via hematoxylin and eosin (H&E) staining. Subsequently, transcriptome sequencing was performed, and JC-1, H2DCFDA, and DHE staining methods were employed to measure mitochondrial function and reactive oxygen species (ROS) levels in PUN-treated cells. Western blotting was used to detect the expression of apoptosis- and cell cycle-related proteins. Next, two gastric cancer cell lines (AGS, HGC27) were selected for in vitro experiments to assess the combined effects of PUN and the copper ionophore elesclomol (ES) (hereafter referred to as ES-Cu, representing the combination of ES and Cu2+). Cell viability was assessed using the MTT assay, and morphological changes were observed under a microscope. Cell proliferation and migration abilities were assessed via colony formation and scratch wound healing assays, respectively. Fluorescence staining was used to examine mitochondrial function and ROS levels in cells co-treated with PUN and ES-Cu. Laser confocal microscopy and Western blotting were employed to determine the oligomerization level of DLAT protein by quantifying soluble and insoluble protein expression, along with the expression of ACO2, ETFDH, FDX1, and LIAS proteins. Molecular docking, molecular dynamics simulations, immunofluorescence staining, and transfection techniques were utilized to confirm the critical role of FDX1 in copper-induced cell death following co-treatment with PUN and ES-Cu. We found that PUN could suppress the viability, proliferation, and migration of gastric cancer cells in a concentration- and time-dependent manner. Subsequent in vivo experiments demonstrated that PUN inhibited tumor growth in nude mice at a safe dosage. Besides, transcriptome sequencing and Western blot analysis revealed that PUN induced cell cycle arrest and apoptosis. Secondly, the cell death mechanism triggered by PUN was closely associated with mitochondrial stress. PUN increased intracellular ROS levels and reduced mitochondrial membrane potential. Transcriptome sequencing and proteomic analysis further revealed molecular changes at both the mRNA and protein levels, with differential gene analysis identifying potential targets and pathways. Moreover, when PUN was combined with ES-Cu, cell viability, proliferation, and migration were all suppressed, while exacerbating mitochondrial dysfunction and elevated oxidative stress. Laser confocal microscopy and Western blotting were used to assess the expression of soluble and insoluble proteins, confirming the oligomerization of the DLAT protein. Western blot also showed that PUN could regulate the expression levels of ACO2, ETFDH, FDX1, and LIAS proteins. Molecular docking, molecular dynamics simulations, and siRNA transfection were performed to confirm the critical role of ferredoxin 1 (FDX1) in copper-induced cell death. Furthermore, when used in combination with chemotherapy drugs, PUN exhibited synergistic inhibitory effects on gastric cancer growth. PUN demonstrates significant antitumor activity both in vivo and in vitro by inducing mitochondrial dysfunction, which subsequently triggers apoptosis. In combination therapy strategies, PUN can work synergistically with chemotherapy drugs to suppress gastric cancer growth. Furthermore, PUN enhances its inhibitory effect on gastric cancer by working synergistically with cuproptosis through targeting FDX1.

Periodontitis is a chronic inflammatory condition affecting billions globally, posing a significant public health challenge due to its high prevalence and associated tooth loss. The inflammatory microenvironment engendered by periodontitis can induce cellular senescence and functional impairment in critical reparative cells, such as periodontal ligament stem cells (PDLSCs), severely compromising their osteogenic differentiation potential and thereby obstructing the regeneration and repair of periodontal tissues, particularly alveolar bone. Although existing fundamental treatments, including subgingival scaling, and surgical interventions can partially manage the disease, they exhibit notable limitations in eradicating deep-seated inflammation and effectively promoting structural bone regeneration. Consequently, there is an urgent need to develop novel biological treatment strategies aimed at reversing the senescent state of PDLSCs and enhancing their regenerative capacity. Flufenamic acid (FFA) is a widely utilized non-steroidal anti-inflammatory drug known for its notable anti-inflammatory and osteogenic properties. It holds significant potential for application in periodontal tissue engineering; however, its precise effects and underlying mechanisms remain inadequately understood. In this investigation, FFA effectively reversed the senescent state of periodontal ligament stem cells (PDLSCs), resulting in a marked down-regulation of pro-inflammatory, cellular senescence, and osteoclast differentiation-related markers, alongside an up-regulation of osteogenic differentiation-related markers. Furthermore, FFA significantly inhibited M1 polarization and osteoclast differentiation activity in macrophages and osteoclast precursor cells. Drug target screening and molecular docking analyses indicated that FFA mitigates PDLSC senescence and enhances their osteogenic capacity through activation of the SIRT1 signaling pathway. Additionally, this study employed the biological effects of M1 macrophage membranes to develop biomimetic hybrid nanovesicles (FFA@M1-LPs) designed to respond to inflammatory microenvironments. These findings suggest that FFA could be a promising new drug for periodontitis treatment and offer insights for developing drug delivery strategies to effectively regenerate periodontal tissue.

High-grade B-cell lymphoma with concurrent MYC and BCL2/BCL6 rearrangements (HGBL-DHL) is a highly aggressive disease that is resistant to conventional first-line immunochemotherapeutic regimens. This resistance necessitates the exploration of innovative therapeutic strategies.

Osteoporosis (OP) is a prevalent degenerative musculoskeletal disorder in middle-aged and elderly populations, characterized by reduced bone mineral density and an increased risk of fragility fractures. Its pathogenesis is closely linked to oxidative stress imbalance; however, effective long-term therapeutic options remain limited. Here, we report that diacerein, a clinically used drug for osteoarthritis, significantly ameliorates OVX (ovariectomy)-induced bone loss and abnormal bone metabolism. Mechanistically, we demonstrate that diacerein activates the transcription and activity of NRF2 (nuclear factor erythroid 2-related factor 2), a central regulator of the antioxidant response, leading to the restoration of cytoprotective protein expression and thereby exerting protective effects against osteoporosis. The beneficial effects of diacerein on bone protection were largely abolished in Nfe2l2 knockout mice, further underscoring the essential role of NRF2. Further investigation revealed that diacerein is metabolized in vivo into rhein, which exerts potent epigenetic activity. The upregulation of NRF2 appears to be primarily mediated through rhein's inhibition of DNA methyltransferases (DNMTs) 1 and 3a. In summary, our study suggests that diacerein, a drug with a well-established safety profile and suitability for long-term use, represents a promising therapeutic candidate for the treatment of osteoporosis.

Chronic stress is associated with inflammatory activation and oxidative stress responses leading to endothelial dysfunction, which promotes the development of atherosclerosis (AS). SGLT2 inhibitors, such as Dapagliflozin (DAPA), exhibit a protective effect against cardiovascular diseases. However, the effects and mechanisms of DAPA on chronic stress-induced AS are largely unknown. The aim of this study was to determine whether DAPA confers a protective effect against chronic stress-induced AS and to elucidate its further molecular mechanisms. The combined high-fat diet-fed and chronic unpredictable mild stress in ApoE-/- mice and lipopolysaccharides- and corticosterone-induced human umbilical vein endothelial cells (HUVECs) were employed to evaluate the antiatherosclerotic effect of DAPA under chronic stress in vivo and in vitro. Histological staining, western blot analysis, siRNA transfection, reactive oxygen species (ROS) staining, and apoptosis assessment were used to investigate the potential mechanisms of DAPA against AS under chronic stress. The results indicate that DAPA significantly improved plaque size and increased plaque stability in the aorta under chronic stress and reduced inflammation and oxidative stress and inhibited apoptosis in the aorta and HUVECs. Chronic stress upregulated regulated in development and DNA damage response 1 (REDD1) expression, which exacerbated cellular inflammation, oxidative stress, and apoptosis levels, leading to endothelial dysfunction. In contrast, DAPA downregulated REDD1 expression and activated the AKT/FoxO1 pathway. In addition, p53 was a transcriptional regulator of REDD1 under chronic stress. More importantly, p53 agonists prevented DAPA from downregulating REDD1 and inhibited AKT/FoxO1 activation, thereby exacerbating chronic stress-induced endothelial dysfunction. These results suggest that DAPA effectively attenuates chronic stress-induced endothelial dysfunction and AS by downregulating REDD1 to activate the AKT/FoxO1 pathway.

Pharmacological enhancement of endoplasmic reticulum (ER) proteostasis is an attractive strategy to mitigate pathology linked to etiologically diverse protein misfolding diseases. However, despite this promise, few compounds have been identified that enhance ER proteostasis through defined mechanisms of action. We previously identified the phenylhydrazone-based compound AA263 as a molecule that promotes adaptive ER proteostasis remodeling through mechanisms including preferential activation of the ATF6 signaling arm of the unfolded protein response (Plate et al., 2016). However, the protein target(s) of AA263 and the potential for further development of this class of ER proteostasis regulators had not been previously explored. Here, we employ chemical proteomics to demonstrate that AA263 covalently targets a subset of ER protein disulfide isomerases, revealing a potential molecular mechanism for the activation of ATF6 afforded by this compound. We then use medicinal chemistry to establish next-generation AA263 analogs showing improved potency and efficacy for ATF6 activation, as compared to the parent compound. Finally, we show that treatment with these AA263 analogs enhances secretory pathway proteostasis to correct the pathologic protein misfolding and trafficking of both a destabilized, disease-associated α1-antitrypsin (A1AT) variant and an epilepsy-associated GABAA receptor variant. These results establish AA263 analogs with enhanced potential for correcting imbalanced ER proteostasis associated with etiologically diverse protein misfolding disorders.

Colorectal cancer (CRC) ranks among the leading causes of cancer-related mortality worldwide. Hydrogen sulfide (H2S) has been found to possess a characteristic of anticancer, which may offer a potential novel treatment for CRC. Here, we discover the potential targets and mechanism of H2S intervention in CRC employing multiomics analysis and experimental validation. The key targets of H2S intervention in CRC were identified by integrating differentially expressed genes (DEGs) from tumor and normal tissues, the CRC-associated genes, and the targets of H2S. The STRING and Cytoscape tools were explored to obtain hub genes. Functional enrichment analysis, assessment of diagnostic and prognostic significance, single-cell datasets, and cell experiments were used to explore the impact of core targets on CRC and the potential mechanism through which H2S exerts regulatory effects on CRC. Our results identified 9250 genes closely linked to CRC from DEGs and CRC-associated genes, 505 targets for H2S, and 322 potential targets of H2S intervention in CRC. Subsequently, five hub genes were filtered, including MAPK1, MAPK3, JUN, ESR1, and AKT1. The 322 common targets were enriched in the cellular stress responses and IL-17 signaling pathway. Additionally, MAPK3 had good diagnostic and prognostic value for CRC. JUN was highly expressed in immune cells. Cell experiments showed that sodium hydrosulfide (NaHS), a donor of H2S, prominently inhibited cell proliferation, promoted cell apoptosis for CRC, and downregulated the expression of MAPK1, MAPK3, AKT1, and JUN. Taken together, this study elucidates the possible genes and therapeutic mechanisms underlying exogenous H2S intervention in CRC, thereby laying a foundation for the further development of H2S-based therapeutic strategies in CRC management.

We identified the characteristics of CBL and CIPK families of Magnolia biondii and the MbCBL4-overexpressed Arabidopsis plants conferred salt tolerance. Calcineurin B-like protein (CBLs) and CBL-interacting protein kinase (CIPKs) are important components of the Ca2⁺-mediated signal pathway. These proteins play a key role in plant growth, development, and response to environmental stress. Magnolia biondii is a woody plant valued for ornamental and medicinal uses and is frequently exposed to abiotic stresses during its growth cycle. Nevertheless, there are still gaps in the study of CBL and CIPK gene families in M. biondii. In this study, 6 CBL and 20 CIPK genes were identified from the M. biondii genome. Phylogenetic analysis divided these genes into 4 CBL and 7 CIPK subgroups, and cross-species comparisons across 34 plants indicated that monocotyledons generally harbor more CBLs/CIPKs than Magnoliaceae. Quantitative real-time PCR (qRT-PCR) analysis revealed that MbCBLs and MbCIPKs showed different transcription levels under drought, cold, and salt stress. Protein-protein interaction assays (Y2H and LCI) verified physical interaction of MbCBL1/MbCIPK18 and MbCBL4/MbCIPK18. Functionally, MbCBL4 overexpression in Arabidopsis conferred enhanced salt tolerance: primary root length and chlorophyll content increased by 2.74-fold and 2.71-fold relative to wild type; fresh weight increased by up to 60%, SOD and CAT activities rose by 47% and 28%, while H₂O₂ and O₂⁻ levels declined by 46% and 38%. These results indicate that MbCBL4 enhances salt tolerance by promoting growth, antioxidant capacity, and reactive oxygen species scavenging. These findings provide important insights into the functional roles of MbCBL and MbCIPK genes and the regulation of MbCBL4 under salt stress.

Particulate matter (PM) poses risks to environmental and human health, yet its toxicity mechanisms in aquatic organisms remain unclear. This study investigated the effects of 2 PM types, a standard reference material (S-PM10, NIST, USA) and particulates from the Mae Moh Power Plant, Thailand (MMPS), on zebrafish embryo development and gene expression. Embryos were exposed to various concentrations, and mortality, hatching rates, and morphological abnormalities were assessed. S-PM10, with irregular morphology and broad particle size, induced developmental defects and reduced hatching. MMPS, characterized by uniform, spherical particles, caused higher mortality. qRT-PCR revealed that S-PM10 significantly upregulated oxidative stress (sod1, gstp2) and apoptosis (bax, casp3a) genes. In contrast, MMPS downregulated oxidative stress markers but upregulated apoptosis-related genes. These results suggest particle morphology and size influence toxicity profiles, S-PM10 triggers developmental disruption, while MMPS induces acute lethality. This study underscores the importance of particle characteristics and molecular responses in evaluating PM toxicity.

Cisplatin resistance remains a major challenge in bladder cancer. Although the tumor suppressor ASPP2 is a critical co-factor for TP53-mediated apoptosis, its role in metabolic reprogramming and cisplatin response remains unclear. This study aimed to delineate the mechanism by which ASPP2 regulates cisplatin sensitivity through metabolic reprogramming. We first assessed the clinical significance of ASPP2 using patient tissues and public databases, finding that its downregulation in bladder cancer is associated with poor patient survival. Through gain- and loss-of-function studies in vitro and in vivo, we further demonstrated that ASPP2 inhibits the mevalonate (MVA) pathway independently of TP53 status, thereby sensitizing cells to cisplatin-induced DNA damage and apoptosis. This chemosensitizing effect was specifically reversed by the addition of MVA pathway metabolites. Moreover, WWP2 was identified as the E3 ubiquitin ligase responsible for ASPP2 degradation via K48-linked ubiquitination. Finally, WWP2 silencing was shown to stabilize ASPP2, suppress the MVA pathway, and synergize with cisplatin to impede tumor growth in mouse models. Overall, the WWP2-ASPP2-MVA pathway axis is identified as a novel driver of cisplatin resistance in bladder cancer. These results establish a mechanistic basis for targeting this axis to restore chemosensitivity, offering a promising therapeutic strategy for recalcitrant disease.

Low temperature combined with low light stress affects off-season pepper cultivation by impairing growth and reducing yield. Betaine acts as osmoregulatory, improving plant stress resistance, but how it mediates m6A methylation remains unknown. This study investigates effects of betaine on low temperature combined with low light conditions by treating pepper seedlings and comparing m6A methylation patterns in the transcriptome, alongside validating methylase gene functions through VIGS and overexpression techniques. Results indicated m6A modification sites affected by betaine are located within coding sequences and 3' untranslated regions under LL stress. Post-treatment with betaine upregulated m6A peaks significantly outnumbered downregulated ones. Transcriptome analysis revealed that betaine enhanced m6A methylation, leading to the degradation of genes like auxin-induced protein AUX28, acid phosphatase-1-like, calmodulin-7, U-box domain-containing protein-21-like, late embryogenesis abundant protein D-34-like, and Protein TAR1. However, it stabilized genes of bHLH87-like, pyrophosphate-energized vacuolar membrane proton pump-like, ABC transporter C family member 4-like, F-box domain-containing protein with hypo-m6A methylation, which are linked to peroxidase synthesis and O-methyltransferase metabolic pathways. Additionally, CaMTC enhanced the tolerence of pepper seedlings to low temperature combined with low light by increasing the m6A methylation level, outlining a regulatory network associated with m6A methylation and betaine. Overall, these findings provides a theoretical framework for the role of betaine in m6A modification under low temperature combined with low light stress, and offers a potential strategy for betaine to enhance plant resistance through the regulation of m6A methyltransferases.

Glutathione (GSH) serves as a redox-active molecule and the predominant non-protein sulfhydryl compound in plants and a critical regulator in alleviating abiotic stress. Our previous research has demonstrated that foliar application of exogenous GSH can enhance the salt tolerance of tomato seedlings. However, the underlying molecular mechanism remains unexplored. In this study, RNA-seq analysis revealed that exogenous GSH significantly influenced plant hormone signal transduction, MAPK signaling pathway, and starch and sucrose metabolism. In addition, the transcription factor SlMYB48 was identified. The expression of SlMYB48 was strongly induced under salt stress but suppressed when GSH was applied simultaneously. Transgenic overexpression (OE) and knockout mutant lines of SlMYB48 were generated and exposed to salt stress, demonstrating that SlMYB48 functioned as a negative regulator of salt tolerance in tomato seedlings. Foliar GSH application increased endogenous GSH content, enhanced the activity and expression of key enzymes in GSH metabolism and the antioxidant system, and reduced ROS accumulation and oxidative injury in OE lines subjected to salt stress. Furthermore, exogenous GSH significantly elevated the expression of starch and sucrose metabolism related genes and increased the corresponding sugar content in OE plants under salt stress. Importantly, GSH application suppressed SlMYB48 expression in the OE lines exposed to salinity. Collectively, these findings indicate that exogenous GSH enhances salt tolerance in tomato seedlings by repressing SlMYB48 expression, thereby modulating the antioxidant system and osmotic adjustment. This study establishes a theoretical framework for elucidating GSH regulated molecular breeding for salt resistance, highlights SlMYB48 breeding potential, and guides practical applications.