Endocrine resistance is frequently encountered in estrogen receptor-positive (ER+) breast cancer, often because of somatic mutations such as neurofibromin 1 (NF1) loss. The mechanisms by which ER-directed proliferation is lost in such cases are unknown, limiting the potential use of additional endocrine treatments. Here, we performed CRISPR-Cas9 knockout (KO) screens and found that nuclear receptor subfamily 2 group F member 2 (NR2F2), an orphan nuclear receptor, was essential for NF1 loss-induced endocrine resistance. Induction of NR2F2 was observed in ER+ cell line models and patient samples and occurred via activation of the mitogen-activated protein kinase (MAPK) pathway upon NF1 loss or other MAPK pathway genetic alterations. Mechanistically, increased NR2F2 orchestrated a repressed ER transcriptional program by repartitioning the ER cistrome, altering the balance of its associated transcriptional coregulators, and modifying global chromatin accessibility. Accordingly, genetic depletion or pharmacologic inhibition of NR2F2 restored sensitivity to hormone therapies in multiple models, including ER+ cell lines, patient-derived xenografts, and patient-derived organoid-based xenografts harboring diverse endocrine-resistance mechanisms such as NF1, AT-rich interactive domain-containing protein 1A (ARID1A), phoshatase and tensin homolog (PTEN) loss, or Kirsten rat sarcoma virus (KRAS) overexpression. Together, these findings underscore NR2F2 as a critical modulator of the hormone response pathway and suggest its inhibition as a promising strategy to overcome endocrine resistance in breast cancer.

Indisulam, a sulfonamide-based compound, is employed as a second-line therapy for NSCLC due to its anti-tumor activity. However, its clinical efficacy is hindered by acquired resistance, the molecular basis of which remains poorly understood. Here, we demonstrate that hypermethylation of RNA-binding protein 39 (RBM39), a specific target of Indisulam, is closely associated with Indisulam resistance. PRMT6 methylates RBM39 at R92. This methylation inhibits Indisulam-induced ubiquitination and proteasomal degradation of RBM39, increases RBM39 protein levels, promotes alternative splicing and expression of proto-oncogenes, and ultimately leads to malignant proliferation and metastasis of NSCLC cells and tumor growth in xenograft mouse models. Inhibiting PRMT6 with MS023 or mutating the RBM39 methylation site enhances Indisulam sensitivity in NSCLC and significantly improves its anti-tumor efficacy. Our findings identify methylated RBM39 as a key biomarker of Indisulam resistance and suggest a potential therapeutic strategy for NSCLC.

Our previous research demonstrated that curcumin suppresses mouse colorectal cancer (CRC) cell CT26 migration and invasion by inhibiting heparanase (HPSE) mRNA expression. To further elucidate the mechanism of curcumin in human CRC treatment, we hypothesized that HPSE plays a pivotal role in human CRC metastasis and that curcumin inhibits this process by downregulating HPSE expression through epigenetic regulation mediated by non-coding RNAs. For further research, human CRC cells were infected with lentivirus to establish overexpression of HPSE cell lines and corresponding negative control cell lines. In vitro and in vivo experiments showed that curcumin inhibited the proliferation, migration, and metastasis of CRC cancer by inhibiting HPSE expression. In the tumor microenvironment, HPSE played an important role in activating the IL-6/STAT5 axis signaling pathway by destructing the extracellular matrix and releasing large number of cytokines, while changing the tumor microenvironment and EMT process, thus promoting tumor metastasis. RNA-seq analysis combined with qRT-PCR results showed that curcumin's inhibition of HPSE expression involved the regulation of non-coding RNAs. Taken together, our results suggested that HPSE promotes CRC metastasis by activating the IL-6/STAT5 signaling axis, disrupting the ECM, releasing cytokines, and altering the tumor microenvironment to facilitate EMT. Curcumin significantly inhibits CRC cell proliferation, migration, and metastasis by downregulating HPSE expression via non-coding RNAs, which related to IL-6/STAT5 axis signal pathways. This research provides a comprehensive understanding of the molecular mechanisms underlying curcumin's anti-CRC effects, emphasizing the role of HPSE and non-coding RNAs in tumor metastasis. These findings pave the way for the development of novel therapeutic strategies targeting HPSE and its regulatory pathways in CRC.

Uterine sarcoma is strongly associated with poor prognosis. However, its treatment options remain limited. Tazemetostat is a potent and selective EZH2 inhibitor with limited clinical application. Entinostat is one of the strong inhibitors for HDAC1 and HDAC3. This study aimed to assess the effect of dual targeting of EZH2 and HDACs on the phenotype of uterine sarcoma cells in both 2D and 3D culture systems. The uterine sarcoma cell line (MES-SA) was treated with varying concentrations of tazemetostat and/or entinostat for 24, 48 and 72 h. For 3D culture conditions, the cells were combined with Matrigel and seeded in V-bottom plates and incubated for 5 days. Cell proliferation, cell cycle progression and apoptosis were evaluated. Additionally, the RNA expression, IHC staining, wound healing assay, DNMT and HDAC activity measurements were conducted. Our data showed that single-inhibitor treatment with entinostat or tazemetostat significantly increased the cytotoxicity and significantly enhanced apoptosis concomitantly. Furthermore, both inhibitors induced cell cycle arrest in 2D and 3D culture conditions. We also demonstrated that entinostat, but not tazemetostat, suppressed the wound healing in the 2D culture. The combination treatment showed a significantly superior effect compared to single-agent treatment. Our studies demonstrate that treatment with either entinostat or tazemetostat alone showed a potent anti-uterine sarcoma effect in 2D and 3D culture conditions. Importantly, the combination of entinostat and tazemetostat produced superior therapeutic effects, suggesting that dual targeting EZH2 and HDACs may provide a promising treatment option for this aggressive cancer.

Misuse and overuse of antibiotics have led to the rapid emergence of antibiotic-resistant superbugs. In addition, evidence is emerging that antibiotic exposure could impose substantial influence on bacterial virulence, but the underlying mechanisms remain poorly understood. Here, we discovered a highly conserved aminoglycoside-responsive regulator, AmgR, that inversely modulates the production of destructive toxins [pyocyanin (PYO) and protease] and the inter-bacterial competition weapon [type VI secretion system (H1-T6SS)], which are the signature virulence factors involved in acute and chronic infections, respectively, in Pseudomonas aeruginosa. We demonstrated that AmgR positively regulates PYO and protease productions by directly activating the transcription of their biosynthetic genes and negatively regulates H1-T6SS indirectly through the quorum sensing regulator PqsR. Importantly, we showed that AmgR can be induced by sub-inhibitory concentrations of aminoglycoside antibiotics to trigger the bacterial chronic-to-acute virulence switch, by promoting P.aeruginosa to withdraw from production of the chronic infection-associated virulence factor H1-T6SS to gear up for generation of acute infection related toxins PYO and protease. This study highlights the risks of improper antibiotic usage not only in elevating antibiotic resistance but also in reprogramming bacterial virulence to exacerbate disease dissemination and acute lethality, providing critical insights for the optimization of antibiotic therapies.

Frequent drug resistance seriously limits the therapeutic efficacy of sorafenib in advanced hepatocellular carcinoma (HCC). Strategies to increase the response to sorafenib are limited, and the underlying mechanism to facilitate such an increase is not entirely understood. Homeobox (HOX) transcript antisense intergenic RNA (HOTAIR) expression is high in HCC, promoting the occurrence and progression of HCC. In this study, we explored the mechanism through which HOTAIR knockdown affects the response of HCC cells to the chemotherapeutic sorafenib.

Quercetin, a well-known antioxidant, plays a crucial role in the response of plants to biotic and abiotic stresses. Triticum urartu is the donor of the A subgenome of common wheat. This study aimed to reveal the mechanism by which quercetin treatment alleviates salt stress in T. urartu. The quercetin treatment resulted in the reduction of Na+ content and enhancement of K+ content in the stressed leaves, while it elevated the Na+ content in the stressed roots. This indicated that under salt stress, quercetin treatment can promote transporting K+ to the leaves and retaining Na+ in the roots. Transcriptome analysis showed that in the roots, quercetin treatment increased the expression level of several genes encoding the rate-limiting enzyme for ethylene biosynthesis, core ethylene signaling proteins, and negative regulators of the core ABA signaling pathway (abscisic acid 8'-hydroxylase and protein phosphatase 2C) under salt stress, revealing that quercetin treatment may induce ethylene signal and suppress ABA signal. We observed that quercetin treatment increased the expression level of several respiration genes in roots under salt stress, including 36 glycolysis genes, 18 mitochondria-related genes, and 4 ATP synthase genes. This displayed that quercetin treatment may enhance the respiration of T. urartu. The enhancement of respiration will provide more energy and carbon sources for salt stress response. Overall, quercetin treatment alleviates the negative effect of salt stress in T. urartu plants via multiple mechanisms, offering potential for improving plant salt tolerance.

During citrus fruit development, exogenous gibberellin (GA) and 6-benzylaminopurine (6-BA, a synthetic cytokinin (CTK)) are both known to promote citrus peel thickness; however, the differences in their regulatory mechanisms on cell wall metabolism in citrus peels remain unclear. In this study, we found that GA treatment significantly increased cell wall polysaccharides in citrus peels, such as pectin and cellulose, whereas 6-BA treatment led to a notable accumulation of lignin. RNA-sequencing data revealed that several fruit ripening-related cell wall degradation genes, such as PME3, PL18, and EXPA1/8, exhibited decreased expression levels in both GA and 6-BA treatments. Additionally, a set of cell wall polysaccharide synthesis genes was upregulated in response to GA treatment but was largely downregulated in 6-BA-treated peels. Conversely, a group of lignin biosynthesis genes was upregulated in 6-BA-treated peels. GA treatment inhibited DELLA proteins (encoded by RGA and GAI) in the GA signaling pathway, whereas 6-BA treatment increased the expression of B-ARRs (ARR1 and ARR2) in the CTK signaling pathway. Furthermore, GA treatment elevated endogenous CTK levels, while 6-BA treatment also enhanced endogenous GA content, suggesting a reciprocal interaction between these two hormonal pathways.

Since the endoplasmic reticulum (ER) stress response is found in almost all tissues, its regulation and downstream factors have been vigorously explored.

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cell carcinoma (RCC), with a rising incidence worldwide. However, the mechanisms by which ccRCC evades immune surveillance remain incompletely understood. Our findings indicate that fumarate hydratase (FH) expression is significantly downregulated in ccRCC, resulting in fumarate accumulation, which is correlated with a poor prognosis in ccRCC patients. RNA sequencing analysis suggests that dimethyl fumarate (DMF), an FDA-approved fumarate analogue, may impact tumor immunity. Our further investigation reveals that both DMF and the FH inhibitor (FHIN1) can promote immune evasion in ccRCC by upregulating PD-L1. Pre-treatment of tumor cells with DMF notably inhibits the cytotoxic effect of T cells. Mechanistically, fumarate induces PD-L1 expression through succination of HIF-1α at C800, facilitating its interaction with importin α3, p300, and PKM2, which promotes HIF-1α nuclear localization and transcriptional activity. Moreover, combining DMF with PD-L1 blockade therapy significantly enhances the efficacy of immunotherapy and prolongs the survival of tumor-bearing mice. Taken together, our study elucidates a mechanism by which FH downregulation promotes immune evasion through the fumarate-HIF-1α/p300/PKM2-PD-L1 axis, providing a novel target, drug, and strategy to improve immunotherapy for ccRCC.

Equine endometrial fibrosis is a leading cause of subfertility in aging mares. Fibrosis is a reparative response involving excess deposition of extracellular matrix (ECM) and increasing tissue stiffness. Augmented rigidity itself can drive fibrosis, by stimulating transition from fibroblasts to myofibroblasts. Myofibroblasts release latent transforming growth factor beta 1 (TGF-β1) from the ECM, thereby activating this profibrotic cytokine. Tissue culture polystyrene (TCP) is commonly used for in vitro experiments. The endometrium, however, is considerably softer than TCP. This study critically evaluated the use of hydrogels versus TCP. Differences in transcript abundance between equine endometrial fibroblasts cultured on TCP and hydrogels of decreasing stiffness (25 kPa to 2 kPa) and their transcriptional response to TGF-β1, were examined. Cells cultured on substrates of varying stiffness exhibited visual variations concerning adherence, morphology, and cell density, besides differences in basal gene expression and transcriptional response to TGF-β1. On stiffer substrates, the smooth muscle genes TAGLN and ACTA2, alongside the transcripts encoding the signaling proteins PDGFB, CCN2, and SERPINE1 were expressed at higher levels. This pattern was also evident for integrin receptor subunit ITGAV, while ITGB5 was expressed at lower levels on stiffer substrates. While ITGB3 demonstrated a response to TGF-β1 exposure independent of stiffness, an increase in transcript abundance of PDGFB, ITGAV, and ITGB5 towards TGF-β1 was only observed on softer hydrogels. The results highlight the importance of stiffness in cellular regulatory patterns, particularly relevant to fibrosis research. We recommend critically reconsidering the use of TCP when designing experiments in vitro.

This study investigates the mechanisms underlying acquired resistance to FGFR tyrosine kinase inhibitor (FGFR-TKI) in gastric cancer (GC), focusing on the interplay between ferroptosis and lipid metabolism of tumor cells.

The phytohormone methyl jasmonate (MeJA) enhances plant cold stress tolerance, but the underlying mechanisms remain elusive. Here, we discovered that MeJA induces a transient Ca2+ influx and elevated cytoplasmic free Ca2+ ([Ca2+]cyt) levels during the watermelon (Citrullus lanatus) response to cold stress. Conversely, silencing jasmonic acid carboxyl methyltransferase (ClJMT), encoding an enzyme that methylates JA to MeJA, led to contrasting effects compared with MeJA application. Upon cold exposure, MeJA rapidly and continuously upregulated 2 Ca2+-permeable channel genes, namely cyclic nucleotide-gated ion channel (ClCNGC) 2 and ClCNGC20. Silencing ClCNGC2 or ClCNGC20 attenuated MeJA-induced Ca2+ influx, [Ca2+]cyt accumulation, C-REPEAT BINDING FACTOR (CBF) pathway activation, and watermelon cold tolerance. Accordingly, ClCNGC2 or ClCNGC20 overexpression increased Ca2+ influx, [Ca2+]cyt levels, and expression of the CBF regulon and improved freezing tolerance in transgenic Arabidopsis thaliana plants. Multiple assays showed that ClCNGC2 and ClCNGC20 do not directly interact. Interestingly, silencing ClCNGC2 or ClCNGC20 abolished MeJA-induced upregulation of ClCNGC20 or ClCNGC2, respectively, in watermelon response to cold, demonstrating their reciprocal activation at the transcriptional level. Collectively, these findings suggest a mutual dependence between ClCNGC2 and ClCNGC20 in mediating MeJA-induced Ca2+ influx followed by [Ca2+]cyt elevation, subsequently activating the CBF pathway and enhancing cold tolerance in plants. This study provides insights into the molecular mechanisms underlying MeJA-mediated plant cold tolerance, holding potential for the breeding or engineering of cold-resistant cucurbit varieties.

Melatonin integrates multiple biological pathways to enhance rice drought tolerance with cooperation of WRKY, bHLH, ERF, MYB and NAC transcription factors, and crosstalk of salicylic acid, ethylene and auxin. Drought is one of the primary environmental threats, and detrimentally affects plant growth and development, impeding crop yield and food quality worldwide. Melatonin has recently emerged as a multifunctional biomolecule with promising aspects in plant stress tolerance. However, the way in which melatonin improves drought tolerance in rice has not been investigated systematically. Here, we demonstrate that melatonin minimized drought effects on rice, resulting in improved germination rate and growth performance. Application of exogenous 200 and 400 µM melatonin can significantly inhibit the accumulation of reactive oxygen species in rice, by enhancing the activity of ROS scavenging enzymes. The accumulation of osmolytes was also stimulated by melatonin to endure with a better preservation of leaf water status in drought-stressed rice. Genome-wide expression profiling by RNA sequencing reveals an increase of oxidoreductase activity, iron ion binding, hydrolase activity, cell wall biogenesis, root development, while a decrease of nitrogen compound metabolism and cellular biosynthesis. Furthermore, melatonin could modulate rice drought response through the cooperation of WRKY, bHLH, ERF, MYB and NAC transcription factors, and modulates salicylic acid, ethylene and auxin pathways. Our finding provides new insights into melatonin-mediated drought tolerance in rice, and facilitates the rational applications in stress management for agricultural, horticultural, and floricultural plants.

Salt stress adversely impacts crop growth and development, resulting in stunted growth and diminished grain yield. Therefore, this study explores the synergetic effects of seed priming with iron nanoparticles (FeNPs) integrated with a genome-wide association study (GWAS) on the phenotypic, biochemical, and agronomic traits of 138 barley accessions under control, salinity stress, and seed iron priming treatments. A normal phenotypic distribution was observed across all accessions under the tested conditions, with significant natural phenotypic variation in response to the treatments. Remarkably, seed priming with FeNPs showed a significant enhancement in superoxide dismutase (SOD) activity and selective modulation of catalase (CAT) and glutathione reductase (GR) activities, indicating a targeted oxidative stress response. Compared to control and salinity stress conditions, priming with FeNPs showed substantial increases in all agronomic traits, including spike length (SL), number of spikelets per spike (NSS), number of grains per spike (NGS), weight of grains per spike (WGS), and thousand kernel weight (TKW), suggesting its potential to mitigate the adverse effects of salinity and promote better crop performance. Based on GWAS analysis, sixteen highly significant marker associations/candidate genes were detected to be associated with antioxidant components. Using quantitative real-time PCR analysis (RT-qPCR), FeNPs seed priming effectively modulates the plant's transcriptional response to salinity stress by balancing rapid gene activation with sustained stress adaptation. This approach mitigates excessive defense responses while promoting long-term stability through controlled upregulation of key genes, such as PP2C, Phosphotransferase, Terpene Synthase Putative, and RWP-RK. The findings support the potential of FeNPs as a biotechnological tool to enhance crop resilience and optimize agronomic performance under adverse environmental conditions.

Grey hair, a common ageing-associated phenomenon in humans, is mainly attributed to the damage of melanocytes and the absence of melanin. Grey hair has long been treated with traditional medicine, and new research has shown that various plant-derived monomers can increase tyrosinase activity and melanogenesis, indicating that they may have therapeutic value in curing grey hair. In this study, we outlined the role of melanin and pigmentation during hair growth and collected various medicinal plant monomers with the potential value of grey hair reversal. Many active ingredients from medicinal plants, such as fraxinol, tribuloside, morin and naringenin, can upregulate melanogenesis and tyrosinase activity through different signalling pathways. Some of them can promote melanosome quantity, maturation and transportation as well. Monomers isolated from medicinal plants may act as stimulators of melanogenesis. Many plant-derived monomers perform as activators that upregulate melanin synthesis and tyrosinase activity through different signalling pathways. They are of great research value for the treatment of hair greying. Moreover, to further improve experimental effect, safety and reliability, a systematic and comprehensive evaluation system needs to be established in the future before studying their clinical efficacy.

Understanding the molecular mechanisms underlying cold and methyl jasmonate (MeJA) responses is vital for improving the cold tolerance of grapes. This study treated 'Pinot noir' plantlets with MeJA, screened key genes in the regulatory pathway using transcriptomics and proteomics analyses, and investigated their regulatory mechanisms under cold stress. The results showed that 50 μmol L-1 MeJA significantly inhibited the growth of grape roots length, increased the endogenous MeJA content and antioxidant enzyme activities, and reduced membrane damage under cold stress. In addition, 50 μmol L-1 MeJA and cold stress treatment greatly increased the number of differential genes and metabolites in the phenylalanine synthesis and hormone signal transduction pathways. The results indicated that VvPAL10, an important gene in the phenylalanine synthesis pathway, significantly improved transgenic Arabidopsis thaliana and grapevine callus tissue tolerance to low temperatures.

Flemingia macrophylla (Willd.) Merr. is commonly used as ethnic medicine in southwestern China, with genistein and genistin being the primary medicinal components. Magnesium has a significant impact on the growth, development, and accumulation of these active ingredients in F. macrophylla. To explore the effects of magnesium on the growth, development, and accumulation of active ingredients in F. macrophylla, tissue culture seedlings of F. macrophylla were treated with six different concentrations of magnesium, namely, T0 (0 g L-1), T0.5 (0.045 g L-1), T1 (0.09 g L-1), T2 (0.18 g L-1), T3 (0.27 g L-1), and T4 (0.36 g L-1), and then subjected to multi-omic analyses. Analysis of agronomic traits revealed that magnesium treatment significantly promoted rooting, whereas magnesium deficiency restricted root growth. Simultaneously, analysis of seedlings using high-performance liquid chromatography showed that 0.27 and 0.36 g L-1 magnesium significantly enhanced genistein synthesis and inhibited glycosylation (genistin). Furthermore, transcriptomic, proteomic, and metabolomic analyses indicated that differentially expressed genes and proteins under varying magnesium concentrations were primarily involved in phenylpropanoid, isoflavone, and flavonoid biosyntheses. Specifically, eight gene-protein pairs related to the synthesis of genistein and genistin were identified. Overall, our study identified essential genes/proteins involved in the synthesis of these two compounds and provided new ways to regulate genistein synthesis in medicinal plants.

Polycystic ovary syndrome (PCOS) is a common endocrine-metabolic disease in women of reproductive age. One of its core pathologies is ovarian granulosa cell (GC) dysfunction, and ferroptosis, as a novel cell death mode dependent on iron ions and lipid peroxidation, may be involved in the PCOS process, but the exact mechanism is unknown. Plumbagin (PLB) shows potential in PCOS treatment due to its antioxidant properties. The present study aimed to elucidate the molecular mechanisms by which PLB ameliorates mitochondrial dysfunction and ferroptosis in PCOS GCs through the YTH N6-methyladenosine RNA binding protein 1/L-type amino acid transporter 1 (YTHDF1/SLC7A5) axis.

Psoriasis is a common chronic inflammatory skin disease characterized by epidermal keratinocyte hyperproliferation and persistent immune activation. Histone deacetylase 3 (HDAC3), a member of the class I HDAC family, plays critical roles in regulating immunity and inflammation. However, its precise expression profile and functional contribution to psoriasis pathogenesis remain poorly defined.