Pyroptosis, a type of programmed cell death, exerts direct influence on inflammatory processes and immune response. A previous study suggests that miltirone exhibits notable anti-tumor activities and has been shown to induce tumor cell pyroptosis. Nevertheless, the therapeutic value of miltirone in kidney renal clear cell carcinoma (KIRC) remains underexplored.

Starch degradation is a fundamental process underlying the development of key postharvest quality attributes in ripening mango fruit; however, its regulatory mechanisms remain incompletely understood. In this study, ethephon (ETH) and 1-methylcyclopropene (1-MCP) were used as ripening modulators to investigate the patterns of starch degradation and its transcriptional regulation in postharvest mango fruit. The results demonstrated that ETH fumigation accelerated the ripening process, while 1-MCP treatment effectively inhibited this process. Transcriptome and RT-qPCR analyses revealed and validated that six starch-degrading genes (MiAMY1.1, MiAMY3, MiBAM3, MiBAM9, MiGWD and MiPWD) were regulated in response to ETH. Furthermore, the transcription factor MiRAP2.11 was identified as a nucleus-localized transcriptional activator of MiBAM3 and MiPWD. Transient transformation assays demonstrated that MiRAP2.11 positively regulated starch degradation, thereby promoting development of ripening and quality attributes. The results indicate that MiRAP2.11 functions as a crucial regulator mediating ethylene-induced starch degradation during mango ripening.

KRASG12C inhibitors, such as sotorasib, show clinical efficacy for non-small cell lung cancer (NSCLC) positive for the G12C mutations of KRAS, but primary and acquired resistance to these drugs remains a clinical problem. In this study, we show that the development of resistance to sotorasib in KRASG12C-positive NSCLC cells was mediated by constitutive activation of EGFR resulting from downregulation of the protein tyrosine phosphatase receptor type R (PTPRR). PTPRR has been identified as a physiologic regulator of ERK signaling in several cancer types. In our study, PTPRR was demonstrated to bind directly to EGFR, facilitating its dephosphorylation on tyrosine residues. Resumption of PTPRR expression in the resistant cells attenuated EGFR phosphorylation and restored sotorasib sensitivity. PTPRR downregulation was associated with gene promoter hypermethylation in the sotorasib-resistant cells and NSCLC tissue samples. Furthermore, low PTPRR expression in tumor specimens was associated with shorter progression-free and overall survival for patients with NSCLC treated with sotorasib. In contrast to sotorasib, high PTPRR expression was associated with a poor response to EGFR tyrosine kinase inhibitors in EGFR-mutated NSCLC, suggesting that PTPRR may broadly regulate EGFR dependence in NSCLC. Finally, dual blockade of KRASG12C and EGFR showed a substantial antitumor effect in a xenograft model of sotorasib-resistant NSCLC. This approach is therefore a rational therapeutic strategy for KRASG12C-positive NSCLC, especially for tumors showing PTPRR downregulation.

Glioblastoma (GBM) is the most aggressive primary brain tumour, associated with a dismal prognosis and an urgent need for innovative therapeutic strategies. To address this challenge, our group developed DMC-GF, a novel brain-targeted curcumin analog engineered to enhance blood-brain barrier permeability by blocking metabolic sites and improving GLUT1 recognition. Although its activity against glioma stem cells has been reported, the direct mechanisms by which DMC-GF acts on GBM cells remain unclear. In this study, we systematically investigated the molecular actions of DMC-GF using phenotypic assays, transcriptome sequencing, and bioinformatics analysis. DMC-GF exerted dose-dependent inhibitory effects on GBM cell proliferation, migration and invasion and concurrently promoted apoptosis, as reflected by reduced Bcl-2 expression, activation of Bax/Caspase-3 and reversal of epithelial-mesenchymal transition (E-cadherin↑, N-cadherin↓, MMP-3↓). Transcriptomic profiling identified THBS1 as a key downstream target, showing marked suppression following DMC-GF treatment. Functional experiments further confirmed that THBS1 knockdown mimics the anti-tumour effects of DMC-GF, whereas THBS1 overexpression partially mitigates its inhibitory actions. Mechanistic studies revealed that DMC-GF suppresses the non-canonical, Smad-independent TGF-β1 pathway by downregulating THBS1, thereby inhibiting PI3K/AKT signalling, as reflected by reduced phosphorylation of AKT, GSK3β and mTOR. Collectively, this work provides the first evidence that DMC-GF exerts anti-GBM effects through modulation of the THBS1/TGF-β1/PI3K-AKT axis. These findings suggest DMC-GF as a compelling brain-targeted therapeutic candidate, providing new mechanistic insights and a potential clinical strategy to overcome therapeutic resistance in GBM.

BRAF inhibitors (BRAFi) have transformed the treatment of BRAF mutant melanoma, but inherent and acquired resistance remains a major barrier to curative outcomes. Resistance arises from interconnected mechanisms: genetic alterations reactivating the MAPK pathway or bypass cascades (e.g., PI3K/AKT/RTK), epigenetic modulation, metabolic reprogramming, and the tumor microenvironment (TME) remodeling. Despite extensive research into these mechanisms, a cohesive framework linking each resistance module to targeted therapeutic strategies is lacking. This review systematically categorizes resistance into intrinsic and acquired subtypes: intrinsic resistance is driven by constitutive molecular traits of BRAF mutant melanoma (e.g., persistent MAPK activation, baseline PI3K/AKT hyperactivity), while acquired resistance emerges via therapeutic pressure-induced genetic mutations, epigenetic shifts, metabolic reprogramming, or TME modifications. For each identified resistance mechanism, we provide a detailed examination of corresponding therapeutic advancements. These encompass the development of next-generation BRAFi, strategically designed combination therapies, epigenetic modulators, immunotherapeutic approaches, and RNA-based therapeutic agents. Furthermore, we underscore the pivotal role of state-of-the-art technologies, such as liquid biopsies, single-cell multi-omics analyses, and artificial intelligence, in facilitating precise resistance monitoring and personalized therapy selection. By integrating these insights, we present a structured, translationally focused framework to guide basic research and clinical decision-making, ultimately advancing precision salvage therapy and trials aimed at preventing or overcoming BRAFi resistance.

This study is aimed to elucidate the role of ubiquitin-specific proteases (USP) 15 in cerebral ischemia-reperfusion injury (CIRI) under the influence of sevoflurane (Sev).

Gastric cancer (GC) is one of the most prevalent gastrointestinal malignancies and continues to pose a significant global health burden. Repurposing clinically approved drugs is a cost-effective strategy for identifying new anticancer agents. In this study, the anticancer effects of linagliptin (LG), a dipeptidyl peptidase-4 (DPP-4) inhibitor, were evaluated in the human gastric cancer cell line HGC-27. LG significantly reduced cell viability in a dose-dependent manner, with a half-maximal inhibitory concentration (IC50) value of 50.29 μM, and increased apoptotic cell death as confirmed by flow cytometry. Gene expression analysis revealed significant downregulation of Smoothened (SMO) and split hand and foot malformation 1 (SHFM1) and upregulation of Sonic hedgehog (SHH), indicating modulation of Hedgehog signaling. In addition, LG treatment resulted in reduced global DNA methylation levels. Molecular docking analyses demonstrated favorable binding affinities between LG and key Hedgehog pathway proteins, supporting a potential mechanistic basis for the observed biological effects. Collectively, these findings suggest that LG exhibits antiproliferative and pro-apoptotic activity in gastric cancer cells and may represent a promising candidate for drug repurposing in gastric cancer therapy.

Programmed cell death (PCD) is a major cause of reduced cell viability following cryopreservation, yet the underlying mechanism remains unclear. In this study, pollen from Paeonia lactiflora was used as the experimental material to investigate the role of the microtubule cytoskeleton in PCD during pollen cryopreservation, which exhibits significant viability decline after cryopreservation. The results showed that post-cryopreservation addition of the microtubule-depolymerizing agent oryzalin significantly decreased pollen viability. This effect was accompanied by the activation of caspase-like proteases, reduced mitochondrial membrane potential, elevated intracellular cytochrome C levels, accumulation of PCD signaling molecules, and ultimately increased apoptosis rates. In contrast, treatment with the microtubule-stabilizing agent paclitaxel exerted the opposite effect. At the transcriptional level, paclitaxel treatment induced 754 differentially expressed genes (DEGs); oryzalin treatment resulted in 575 DEGs, a total of 63 DEGs were shared between the two treatments. At the protein level, paclitaxel treatment yielded 262 differentially expressed proteins (DEPs), while oryzalin treatment led to 270 DEPs, with 100 DEPs overlapping between the two groups. Integrated transcriptomic and proteomic analyses revealed that these DEGs and DEPs were significantly enriched in two key pathways: cysteine and methionine metabolism, and protein processing in the endoplasmic reticulum. Notably, heat shock proteins were prominently expressed at both the transcriptional and protein levels in the endoplasmic reticulum protein processing pathway, while malate dehydrogenase played an extremely critical role in cysteine and methionine metabolism pathway. Collectively, these findings indicate that the microtubule cytoskeleton is involved in regulating PCD during pollen cryopreservation, with cysteine and methionine metabolism and endoplasmic reticulum protein processing serving as the core pathways.

Dengue virus (DENV) remains a major global health concern without effective treatments. Previously, we identified sulfonyl anthranilic acid (SAA) derivatives (compounds 1 and 2) as potent pan-DENV inhibitors, likely targeting a primate-specific factor. Here, mass spectrometry-based target deconvolution revealed that SAA compounds downregulate ribosomal protein expression, some of which are essential for DENV replication, as confirmed by siRNA-knockdown studies. This novel mechanism aligns with the broad-spectrum antiviral activity of compounds 1 and 2. Moreover, compound 1 was also effective against the Zika virus in a human brain organoid model. The subsequent medicinal chemistry optimization process resulted in the identification of compound 7, which demonstrated an EC50 value of 50 nM against DENV-2, promising broad-spectrum potential and favorable in vitro ADME properties. Further studies indicated that these compounds modulate the 5'-terminal oligopyrimidine (5'-TOP) motif in ribosomal mRNAs. These findings open a new avenue for antiviral development by targeting a previously unexplored host pathway.

As an inhibitor of DNA methyltransferases (DNMTs) and an anti-cancer drug, 5-aza-cytidine (5-azaC)'s many effects on gene expression remain unclear. Here, we show that 5-azaC treatment of cultured GH3 pituitary tumor cells increases relative usage of genomic terminal exons (GTEs) across the transcriptome. This effect is largely achieved by shifting mRNA polyadenylation from proximal poly(A) sites to GTEs, which harbor a more optimal consensus motif of poly(A) signals. Consistent with this shift, 5-azaC upregulates the mRNA anti-termination factors Scaf4 and Scaf8 while downregulating the early termination enhancer E2f2. In MOLM-13 leukemia cells, 5-azaC similarly promotes the production of full-length transcripts and regulates alternative polyadenylation factors, some of which are in the same direction as observed in GH3 cells. Moreover, PCF11, a factor known to promote proximal poly(A) site usage, is upregulated in both cell lines, suggesting a homeostatic response by these cells to counteract transcript lengthening during 5-azaC treatment. Together, these findings uncover a previously unknown effect of 5-azaC on gene expression: directional promotion of terminal polyadenylation site usage, driving a transcriptome-wide switch from shortened to full-length mRNAs in tumor or cancer cells and consequently altering the alternative usage of multiple 3' exons.

Current toxicology and cancer biology investigations have focused on developing alternative models that better recapitulate the in vivo architecture of tissues and organs. The present study evaluated the anticancer effects of the flavone cirsimarin, which presented successful antitumor activity on breast tumor cells. We assessed the impact of flavone on cell viability, proliferation, and migration, as well as on DNA integrity and modulation of related cellular pathways. In the 2D model, cirsimarin reduced cell viability at concentrations ≥ 80 μM after 24 h of treatment (resazurin assay), selectively in A549 cells compared to MRC-5 non-tumor cells. Apoptosis was induced at concentrations ≥ 40 μM, and clonogenicity was reduced by approximately 50% only at 160 μM. In the wound healing assay, cirsimarin (1-80 μM) completely inhibited cell migration and induced DNA damage (comet assay). These apoptotic and anti-migratory effects were associated with the downregulation of key genes involved in cell proliferation, death, and extracellular matrix remodeling, including TNF-α (0.32-fold), TP53 (0.17-fold), MMP-2 (0.18-fold), MMP-9 (0.43-fold), and MMP-11 (0.04-fold), as revealed by RT-qPCR analysis. In the 3D model, after 216 h of treatment, cirsimarin reduced cell viability (≥ 40 μM) and spheroid area (≥ 80 μM) while antimigratory effects were observed only in the highest concentration evaluated (160 μM). These findings could indicate a potential reduction in lung tumor growth and metastasis, warranting further investigation, particularly of the antimetastatic effect of this flavone.

Alfalfa (Medicago sativa L.) is a globally important forage legume and the most widely cultivated sown pasture species in China. Drought, as one of the most common abiotic stresses, limits alfalfa growth and development. Nitric oxide (NO), a key signaling molecule, plays an essential role in plant growth, development, and responses to various abiotic stresses. In this study, exogenous NO was applied to alfalfa seedlings under drought stress, followed by physiological and transcriptomic analyses. The results showed that sodium nitroprusside (SNP)-treated alfalfa seedlings grew better than untreated controls (CK), with improved leaf tissue structure. Meanwhile, SNP treatment increased proline content, reduced malondialdehyde accumulation, and enhanced hydroxyl radical scavenging capacity. Under drought stress, lignin content increased in alfalfa seedlings. Following exogenous NO application, lignin content in alfalfa seedlings further increased. RNA-Seq analysis identified 20,183 differentially expressed genes (DEGs) in alfalfa seedlings treated with PEG, SNP, or PEG + SNP. KEGG enrichment analysis indicated that the DEGs under drought stress were involved in the phenylpropanoid biosynthesis pathway, which regulates lignin biosynthesis and enhances drought tolerance. GO enrichment analysis revealed that these DEGs participated in the response to water deprivation, thereby modulating drought stress tolerance and improving drought resistance. Furthermore, we assessed the transcript-level changes in genes induced by phenylpropanoid biosynthesis in alfalfa. Among them, 124 DEGs were identified as participating in phenylpropanoid biosynthesis, including 10 up-regulated DEGs (three of which encode key enzymes associated with lignin synthesis), while the remaining DEGs were down-regulated. These findings provide new insights into the transcriptomic mechanisms of SNP-mediated drought adaptation in alfalfa and reveal key pathways contributing to drought tolerance in this species.

Large-scale randomized trials have found that multivitamin-multimineral (MVM) supplements and cocoa flavanols may benefit several age-related chronic conditions among older adults, but it remains unclear whether these two supplements directly slow the biological aging process. This prespecified ancillary study evaluated the 2-year effect of a daily MVM (Centrum Silver) and cocoa extract (500 mg cocoa flavanols per day, including 80 mg (-)-epicatechin) on five DNA methylation measures of biological aging (PCHannum, PCHorvath, PCPhenoAge, PCGrimAge and DunedinPACE) among 958 participants (482 women and 476 men) in the COcoa Supplement and Multivitamin Outcomes Study (COSMOS). Compared with placebo, daily MVM supplementation modestly reduced the rate of increase of second-generation epigenetic clocks, with a between-group difference in yearly change of -0.113 years (95% confidence interval (CI) -0.205 to -0.020; P = 0.017) for PCGrimAge and -0.214 years (-0.410 to -0.019; P = 0.032) for PCPhenoAge. MVM had a stronger effect on PCGrimAge among those with accelerated biological aging at baseline (-0.236 [-0.380 to -0.091]) compared with those with normal or decelerated biological aging (-0.013 [-0.130 to 0.104]; P = 0.018 for interaction). Cocoa extract did not have an effect on the five epigenetic clocks tested. Although the statistically significant but small effects of daily MVM supplementation on slowing biological aging are encouraging, additional studies are needed to determine the clinical relevance of daily MVM supplementation on epigenetic clocks and whether such effects can help explain the beneficial effects of MVM supplementation on aging-related chronic conditions.

Polycystic ovary syndrome (PCOS) is associated with ovarian granulosa cell dysfunction. Ferroptosis, a regulated cell death driven by lipid peroxidation, represents a novel pathological mechanism. Hypermethylation of the glutathione peroxidase 4 (GPX4) promoter may contribute to its suppression. While glucagon-like peptide-1 receptor agonists (GLP-1 RAs) improve metabolic features of PCOS, their direct effects on ovarian ferroptosis and the underlying epigenetic mechanisms are unclear. To explore the therapeutic potential of GLP-1RAs across PCOS phenotypes, we employed a hyperandrogenism-induced rat model and a letrozole plus high-fat diet mouse model, treating them with exenatide or semaglutide, respectively. Phenotypic assessment included estrous cycle monitoring, ovarian histology, and serum hormone profiling. Ferroptosis was evaluated using a multi-parametric approach, including iron deposition (Perls' staining), lipid peroxidation (MDA), redox status (GSH/GSSG), ultrastructural analysis (TEM), and expression of key regulators. The methylation status of the GPX4 promoter was analyzed by methylation-specific PCR (MSP) and bisulfite sequencing (BSP), alongside the expression of related epigenetic modifiers (DNMTs, TET1). In vitro studies using DHT-stimulated primary granulosa cells further validated the semaglutide effects. GLP-1 RA exenatide alleviated the polycystic ovarian morphology in rats with PCOS, semaglutide treatment not only alleviated PCOS phenotypes but also reversed ovarian ferroptosis markers, restored GPX4 expression, and reduced the GPX4 promoter hypermethylation and DNMTs levels, with efficacy comparable to 5-azacytidine. In vitro, semaglutide corrected DHT-induced GPX4 hypermethylation and ferroptosis in granulosa cells. This study demonstrates that semaglutide alleviates PCOS phenotypes and reverses ovarian granulosa cell ferroptosis. These beneficial effects may be related to the alleviation of GPX4 promoter hypermethylation. Our findings extend the therapeutic rationale for semaglutide in PCOS beyond metabolic benefits, suggesting potential direct ovarian protection via epigenetic modulation.

Multiple myeloma (MM) is an incurable plasma cell neoplasm that is highly reliant on endoplasmic reticulum-associated degradation (ERAD) to maintain protein homeostasis. Disrupting ERAD has been proposed as a therapeutic strategy to overcome proteasome inhibitor resistance; however, the identification of novel inhibitors has been limited. To address this, we conducted a cell-based high-throughput screen using the FDA repurposing library and identified omaveloxolone (RTA408) as a potent ERAD inhibitor that selectively impairs the degradation of ER luminal and membrane substrates, without affecting the degradation of key cytosolic proteins that are implicated in disease relapse. Surprisingly, although ER stress response pathways are activated after ERAD inhibition in MM, we find that apoptosis is mediated by altered lipid raft organization, leading to aberrant activation of the death-inducing signaling complex (DISC) and caspase 8 in the extrinsic apoptotic pathway. Notably, ERAD inhibition by RTA408 is cytotoxic to primary malignant plasma cells, including those resistant to proteasome inhibitors, and demonstrates in vivo anti-myeloma activity. Our findings establish a novel ERAD inhibitor, which is a valuable tool to dissect ERAD biology, and provide pre-clinical evidence for RTA408 as a therapeutic agent in MM.

The treatment of colorectal cancer (CRC) remains challenging due to chemotherapy resistance and genetic heterogeneity. Indole-3-lactic acid (ILA), a tryptophan metabolite derived from gut microbiota, exhibits promising anti-inflammatory and anticancer properties; however, its specific molecular targets and regulatory mechanisms in CRC remain poorly understood. In this study, we combined network pharmacology and machine learning with molecular docking to identify candidate targets and pathways for ILA in CRC. We identified 39 ILA-CRC common targets, ultimately identifying four hub genes through the intersection of machine learning models. Validation in independent GEO datasets confirmed significant differential expression of these genes in CRC tissues. Functional enrichment analyses linked these genes to the PPAR, PI3K-AKT, and IL-17 signaling pathways, and gene set enrichment analysis further implicated ascorbate and aldarate metabolism, DNA replication, and fatty acid metabolism. Immune infiltration analysis indicated associations between hub gene expression and immune cell populations, including mast cells, neutrophils, and macrophages, suggesting potential involvement in the tumor immune microenvironment. Molecular docking supported favorable binding of ILA to all four hub proteins, and 100-ns molecular dynamics simulations specifically validated the dynamic stability of the ILA-HMOX1 complex. In conclusion, these results highlight EPHA2, HMOX1, MMP3, and PARP1 as candidate targets and suggest that ILA may influence CRC-related signaling, metabolic programs, and immune contexture, providing a theoretical foundation for developing gut microbiota-derived metabolites as novel anticancer strategies.

Glucocorticoids (GCs) are steroid hormones that bind to the glucocorticoid receptor (GR) as ligands to initiate systemic anti-inflammatory effects. GCs are commonly administered alongside chemotherapy to reduce treatment-related side effects in breast cancer patients. However, GC administration has been shown to promote metastasis in breast cancer. In this study, we used quantitative mass-spectrometry-based approaches to analyze proteome and phosphoproteome of three breast cancer cell lines following treatment of a clinically approved synthetic GC, dexamethasone (Dex). By comparing MCF7, MDA-MB-231, and MDA-MB-436 cells, we suggest that the level of GR significantly affects Dex-mediated responses. Additionally, we identify noncanonical transcription factors (TFs) and kinases that are regulated by GR in different cell lines. Together, our data present Dex-induced protein modulations and modifications involving several TFs and kinases that regulate cytoskeletal remodeling and migration in breast cancer cell lines. These findings highlight the need for careful consideration of GC use in breast cancer therapy and identify potential molecular targets for mitigating adverse effects.

MsWIP3, a C2H2-type zinc finger transcription factor in alfalfa, plays a key role in enhancing saline-alkali stress tolerance by regulating stress responses and reducing oxidative damage. Saline-alkali stress severely restricts plant growth and yield worldwide, particularly in the Songnen Plain of northeastern China, where alkaline salts with high pH values, mainly NaHCO₃, are a major constraint on agriculture and forage production. C2H2-type zinc finger proteins (C2H2-ZFPs) are important transcription factors involved in plant responses to multiple environmental stresses. However, their roles in alfalfa (Medicago sativa) adaptation to saline-alkali stress remain poorly understood. We integrated transcriptome (RNA-seq) analysis of alfalfa under NaHCO₃ treatment with Weighted Gene Co-expression Network Analysis (WGCNA) to identify stress-responsive transcription factors. MsWIP3, a WIP subfamily member, contains four conserved zinc finger domains and the characteristic "WIP" motif. Subcellular localization and transcriptional activity assays were performed, and the function of MsWIP3 was evaluated through heterologous expression in yeast and Nicotiana benthamiana. MsWIP3 localized to the nucleus and exhibited transcriptional repression activity. Overexpression of MsWIP3 suppressed the growth of yeast and tobacco but significantly enhanced NaHCO₃ tolerance in transgenic tobacco, as evidenced by reduced PSII photoinhibition and lower oxidative damage from reactive oxygen species. Our findings indicate that MsWIP3 plays a role in the saline-alkali stress response of alfalfa. This study contributes to our understanding of the molecular mechanisms of stress adaptation and identifies a potential target for the development of salt-alkali tolerant alfalfa varieties.

Cell surface receptors play vital roles in cancer growth and metastasis. Integrin αvβ3 is overexpressed in various cancer cells and interacts with different growth factors to stimulate cancer progression. Thyroid hormone binds to αvβ3 to activate signal transduction and cell proliferation. However, thyroxine (T4) deaminated analogue, tetraiodothyronine (tetrac), competes for the binding on integrin and inhibits cancer cell growth and metastasis. The current study investigated the pathogenic role of integrin αvβ3 and the potential of a novel therapeutic strategy targeted to integrin αvβ3. Pathogenetic studies of clinical samples revealed integrin αvβ3 cross-talked with EGFR and downstream signal transduction networks affected by thyroid hormone and EGF related to the progression of cholangiocarcinoma malignancy. Thyroxine and EGF stimulated PD-Ligand 1 (PD-L1) expression and cancer growth in cholangiocarcinoma. The thyroxine-induced PD-L1 accumulated in the nuclei and colocalized with p300. Alternatively, EGF increased cytosolic PD-L1 and nuclear accumulation of β-catenin. Targeting integrin αvβ3 with lipo-tetrac and its Dox-derivative induced anti-proliferation in vitro and in the xenografted animal model. Our research provides a fundamental understanding of the therapeutic role of integrin αvβ3 and the potential therapeutic approach in cholangiocarcinoma treatment.

Triple-negative breast cancer (TNBC) is an aggressive subtype lacking targetable hormone receptors, making conventional chemotherapy the primary treatment option, despite its associated toxicity and potential for drug resistance. Melatonin, a natural hormone with anticancer and immunomodulatory properties, has shown promise in multiple cancers; however, its role in TNBC remains unclear.