Metastatic breast cancer (BC) is the main cause of cancer-related mortality in women worldwide. HR + /HER2- BC patients are treated with endocrine therapy (ET), but therapeutic resistance is common. The combination of cyclin-dependent kinase 4/6 inhibitors (CDK4/6i) with ET was approved for metastatic BC patients and extended the median progression-free survival to 24 months. This therapy is not always effective, and in every patient, resistance ultimately occurs, but the underlying resistance mechanisms remain unclear. To address this gap, we explored circulating tumour cells (CTCs) as biomarkers to assess treatment response and resistance in metastatic HR + /HER2- BC patients receiving CDK4/6i plus ET.

The incidence of atherosclerosis markedly rises following menopause. Our previous findings demonstrated that elevated follicle-stimulating hormone (FSH) levels in postmenopausal women accelerate atherosclerosis progression. Plaque instability, the fundamental pathological factor in acute coronary syndrome, primarily results from vascular embolism due to plaque rupture. Recent evidence highlights that endothelial-to-mesenchymal transition (EndMT) exacerbates plaque instability, although the link between FSH and EndMT has not been fully established. This investigation sought to explore the possible influence of FSH in modulating EndMT.

Seed germination represents a pivotal phase in crop production, exhibiting pronounced sensitivity to abiotic stresses. In this study, wheat seeds of the 'Ningchun 4' variety were subjected to treatments involving zinc (Zn) chloride and iron (Fe) chloride, both individually and in combination. The impacts of these treatments on Fe and Zn accumulation, starch mobilization, antioxidant responses, and nitric oxide (NO) metabolism during seed germination were thoroughly examined. Individual application of Fe or Zn significantly inhibited and delayed wheat seed germination, which was accompanied by elevated levels of starch, sucrose, and soluble sugars, as well as increased reactive oxygen species and malondialdehyde concentrations. Concurrently, total amylase and α-amylase activities were downregulated, while antioxidant enzyme activities and the expression of TaCAT, TaAPX, and TaGR were upregulated. Seeds treated solely with Fe exhibited excessive Fe accumulation, heightened Fe2+ content, and diminished Zn content. Conversely, these trends were reversed in seeds treated with Zn alone. Furthermore, reduced NO levels were associated with downregulated nitrate reductase and nitric oxide synthase activities, alongside decreased expression of their corresponding genes in response to Fe exposure. Notably, the above effects induced by Zn alone were less severe compared to those induced by Fe stress. Importantly, the addition of Zn (100 µM or 250 µM) significantly alleviated the detrimental effects of Fe on several parameters in germinating seeds. The results from NO fluorescent probe staining corroborated the quantitative NO measurements across different treatments. In conclusion, an appropriate concentration of Zn effectively promoted the germination of Fe-stressed wheat seeds by mitigating Fe accumulation, attenuating oxidative damage, and enhancing starch mobilization during seed germination.

Further study of the mechanism of glucocorticoid (GC)-induced osteoblast (OB) apoptosis is highly important for the prevention and treatment of GC-induced osteoporosis and osteonecrosis. Serine/arginine-rich splicing factor 1 (Srsf1) expression was downregulated in a dose-dependent manner during GC-induced OB apoptosis. Knockdown of Srsf1 significantly promotes GC-induced OB apoptosis, while overexpression of Srsf1 significantly inhibits GC-induced OB apoptosis. Mechanistically, GC induces the up-regulation of histone deacetylase 4 (Hdac4) in OB, and inhibits the expression of transcription activator forkhead box C1 (Foxc1) by reducing the levels of histone H3 lysine 9 acetylation (H3K9ac) and H3K27ac in the promoter region of Foxc1, thereby down-regulating Srsf1. Next, SRSF1 regulates GC-induced OB apoptosis by regulating Bcl-2 modifying factor (Bmf) alternative splicing. From the perspective of alternative splicing, this study demonstrates that Srsf1 and its regulatory mechanism may serve as a new target for the prevention and treatment of GC-induced osteoporosis and osteonecrosis.

Cadmium (Cd) is a toxic heavy metal whose contamination in soil threatens food safety, agricultural production, and human health. To date, phytoremediation is a low-cost and environmentally friendly method for eliminating Cd contamination. In this study, we report a gene from ramie (Boehmeria nivea) that encodes a dehydration responsive element binding (DREB) factor associated with plant tolerance to Cd, namely BnDREB1. The open reading frame of BnDREB1 comprises 873 bp encoding 290 amino acids and includes a characteristic AP2 domain. Its cloned promoter sequence contains various hormone and stress responsive elements. Quantitative RT-PCR analysis showed that BnDREB1 is expressed in different organs of ramie. Treatments with polyethylene glycol (PEG), abscisic acid (ABA), and Cd upregulated the expression of BnDREB1. Confocal microscopic analysis revealed that BnDREB1 is mainly localized in the nucleus. Overexpression of BnDREB1 in Arabidopsis thaliana increased the tolerance of transgenic plants to Cd, thereby protecting plant growth from its toxicity. Biochemical analysis revealed that overexpression of BnDREB1 reduced the levels of Cd induced malonaldehyde and hydrogen peroxide, inhibited the reduction of Cd caused soluble protein contents, increased the Cd accumulation, and enhanced Cd translocation in transgenic plants. Taken together, these findings suggest that BnDREB1 is an appropriate candidate gene for phytoremediation of Cd-contaminated soil .

Bladder cancer is a major global health concern with high incidence and mortality rates. Both muscle-invasive bladder cancer (MIBC) and recurrent non-muscle-invasive bladder cancer (NMIBC) present significant challenges in treatment. Apigenin, a naturally occurring flavonoid, has shown promise in inhibiting the growth of bladder cancer cells, however, its therapeutic mechanism remains unclear. Single-cell RNA sequencing (scRNA-seq) data analysis and drug target screening were performed. Differentially expressed genes (DEGs) and potential therapeutic targets of apigenin were identified. Molecular docking was utilized to evaluate the binding affinity between apigenin and VEGF-β. In vitro assays were conducted to evaluate the association of VEGF-β and apigenin. Drug target screening identified 51 common targets between apigenin and bladder cancer, with VEGF-β emerging as a dominant gene. Molecular docking confirmed a high binding affinity between apigenin and VEGF-β. VEGF-β was significantly upregulated in fibroblasts from recurrent bladder cancer, correlating with increased tumor malignancy. Enhanced cell communication in VEGF-β-positive fibroblasts contributed to tumor progression. In vitro experiments demonstrated that VEGF-β promotes tumor cell proliferation, migration, and invasion. Apigenin significantly inhibits bladder cancer progression by targeting VEGF-β. The upregulation of VEGF-β in fibroblasts from recurrent bladder cancer highlights its potential as a diagnostic marker and therapeutic target. This study underscores the promise of apigenin as a chemopreventive and therapeutic agent for recurrent bladder cancer.

Huayu Wan (HYW), a traditional Chinese medicine prescription widely used in the clinical treatment of advanced lung cancer, has been clinically proven to effectively inhibit the progression of pulmonary tumors and improve patients' quality of life. However, its specific components and potential anti-cancer molecular mechanisms remain unclear.

Acute myeloid leukemia is a hematological malignancy characterized by acquired genomic aberrations. Mutations in the FMS-like tyrosine kinase 3 gene cause constitutive activation of downstream signaling pathways, thereby driving disease progression and conferring a poor prognosis. Gilteritinib, a tyrosine kinase inhibitor, is a promising treatment for FMS-like tyrosine kinase 3-mutated acute myeloid leukemia. However, gilteritinib resistance remains a significant concern, and its underlying mechanisms are not yet understood.

The emergence of carbapenem resistance in gram-negative bacteria such as Escherichia coli is one of the world's most urgent public health problems. E. coli, which encounter a diverse range of niches in host can rapidly adapt to the changes in surrounding environment by coordinating their behavior via production, release and detection of signal molecules called autoinducers through a cell density dependent communication system known as quorum sensing. Here, in this study we investigated whether imipenem, and acyl homoserine lactone quorum sensing signal molecules influence the transcriptional response within lsr and lsrRK operon which are associated with auto inducer-2 mediated quorum sensing in E. coli. Two E. coli isolates carrying blaNDM were treated with 10% SDS for 20 consecutive days, resulting in the successful elimination of the blaNDM encoding plasmid from one isolate. Plasmid was extracted from the isolate and was transformed into recipient E. coli DH5α by electroporation. The native type, plasmid-cured type, transformant, and E. coli DH5α were allowed to grow under eight different inducing conditions and the transcriptional responses of lsr and lsrRK operons were studied by quantitative real-time PCR method.

Triterpenoids are well-known pharmacological components of Sanghuangporus fungi, such as Sanghuangporus lonicericola. This study investigates the inductive effects of paclobutrazol (PBZ) on triterpenoid biosynthesis in the submerged fermentation of S. lonicericola and explores the induction mechanisms via multi-omics and genetic methods. The addition of 100 mg/L PBZ significantly increases the triterpenoid yield by 151.39%. A total of 29 triterpenoids are tentatively identified, of which 18 are newly presented only under PBZ induction. Moreover, 30 genes involved in the MVA pathway and 31 genes encoding cytochrome P450 monooxygenases assumed to be responsible for decoration are identified. Finally, a MYB transcription factor (SlMYB) is identified and found to be downregulated under paclobutrazol induction. Genetic manipulation of SlMYB demonstrates its negative regulatory effect on four putative target genes, including ACAT, MVD, IDI, and FDPS. Electrophoretic mobility gel shift assays verify the direct interactions with the promoters of MVD, IDI, and FDPS. Taken together, PBZ acts as an effective inducer of triterpenoid biosynthesis in S. lonicericola, and the transcription factor SlMYB is negatively regulated.

Gray mold, caused by Botrytis cinerea, poses an escalating threat to ginseng production in China. Excessive application of chemical fungicides has resulted in severe resistance development. To elucidate the resistance mechanism of B. cinerea to pyrimethanil, transcriptomic comparisons were conducted between resistant (HRG21) and sensitive (FSG43) strains following exposure to pyrimethanil for 2 and 6 h. GO and KEGG analyses identified differential expression of genes associated with ABC and MFS transporters as well as methionine biosynthesis. qRT-PCR validation confirmed a marked upregulation of ABC (BcatrA, BcatrB, BcatrD, BcatrO, and Bmr3) and MFS (Bchex1 and BcmfsM2) transporter genes in HRG21, whereas in FSG43, ABC (BcatrA, BcatrB, BcatrD, and BcatrO) and MFS (Bchex1) transporter genes were downregulated. No significant transcriptional changes were observed in Mrr1, a transcription factor gene, following pyrimethanil exposure. Molecular docking analysis identified potential binding sites and interaction targets for five key transporters (BcatrB, BcatrA, BcatrD, Bmr3, and Bchex1). Furthermore, exogenous methionine supplementation partially mitigated pyrimethanil toxicity in FSG43. However, amino acid sequences of enzymes involved in methionine biosynthesis (BcmetC, BcStr2, Bcmet2, Bcmxr1, and Bcmxr2) exhibited no variation between FSG43 and HRG21. The results indicate that BcatrB plays a central role in pyrimethanil resistance, while methionine contributes only marginally to resistance mechanisms.

Gemcitabine combined with cisplatin is the first-line chemotherapy for advanced cholangiocarcinoma, but drug resistance remains a challenge, leading to unsatisfactory therapeutic effect. Here, we elucidate the possibility of chemotherapy regimens sensitized by inhibiting succinylation in patients with cholangiocarcinoma from the perspective of post-translational modification. Our omics analysis reveals that succinylation of PDHA1 lysine 83, a key enzyme in the tricarboxylic acid cycle, alters PDH enzyme activity, modulates metabolic flux, and leads to alpha-ketoglutaric acid accumulation in the tumor microenvironment. This process activates the OXGR1 receptor on macrophages, triggering MAPK signaling and inhibiting MHC-II antigen presentation, which promotes immune escape and tumor progression. Moreover, we show that inhibiting PDHA1 succinylation with CPI-613 enhances the efficacy of gemcitabine and cisplatin. Targeting PDHA1 succinylation may be a promising strategy to improve treatment outcomes in cholangiocarcinoma and warrants further clinical exploration.

FDA-approved KRASG12C inhibitors, like Sotorasib, target G12C-mutated KRAS in NSCLC. However, issues with insensitivity and drug resistance have emerged, requiring the development of new combination therapies to overcome these limitations. USP47 has been identified as a regulator of cancer-related signaling pathways such as Wnt, Hippo, and p53. However, its role in the KRAS signaling pathway remains largely unexplored and USP47 inhibitors are less developed than those targeting its homolog, USP7. Here, we identify USP47 as a novel therapeutic target in KRASG12C-mutated NSCLC and report K-552, a selective USP47 inhibitor, as a potential treatment strategy. We demonstrate that USP47 stabilizes c-Myc by preventing its proteasomal degradation through deubiquitination, thereby promoting NSCLC cell proliferation. Additionally, the compound K-552, a USP47 inhibitor identified through virtual screening, effectively destabilizes c-Myc and inhibits KRASG12C-mutated NSCLC cell proliferation. Furthermore, USP47 inhibition-either by siRNA knockdown or K-552 treatment-enhances the efficacy of Sotorasib in vitro and in vivo. Together, our findings establish USP47 as a promising therapeutic target in KRASG12C-mutated NSCLC and introduce K-552 as a USP47 inhibitor with potential for combination therapy with KRASG12C inhibitors.

Targeting ubiquitin E3 ligases is therapeutically attractive; however, the absence of an active-site pocket impedes computational approaches for identifying inhibitors. In a large, unbiased biochemical screen, we discover inhibitors that bind a cryptic cavity distant from the catalytic cysteine of the homologous to E6-associated protein C terminus domain (HECT) E3 ligase, SMAD ubiquitin regulatory factor 1 (SMURF1). Structural and biochemical analyses and engineered escape mutants revealed that these inhibitors restrict an essential catalytic motion by extending an α helix over a conserved glycine hinge. SMURF1 levels are increased in pulmonary arterial hypertension (PAH), a disease caused by mutation of bone morphogenetic protein receptor-2 (BMPR2). We demonstrated that SMURF1 inhibition prevented BMPR2 ubiquitylation, normalized bone morphogenetic protein (BMP) signaling, restored pulmonary vascular cell homeostasis, and reversed pathology in established experimental PAH. Leveraging this deep mechanistic understanding, we undertook an in silico machine-learning-based screen to identify inhibitors of the prototypic HECT E6AP and confirmed glycine-hinge-dependent allosteric activity in vitro. Inhibiting HECTs and other glycine-hinge proteins opens a new druggable space.

Trained immunity refers to memory-like responses of innate immune cells when they re-encounter pathogenic stimuli. Bacillus Calmette-Guérin (BCG) vaccination implies enhanced antiviral immunity, whereas the underlying mechanisms remain unclear. Herein, we have uncovered elevated expression of low-density lipoprotein receptor (LDLR) on BCG-trained macrophages with robust type I interferon (IFNI) production and antiviral effects both in vivo and in vitro. Consequently, cholesterol is accumulated in BCG-trained macrophages, leading to the augmentation of NFE2L1 expression and the formation of NFE2L1/IRAK1/TRIM25 complex where TRIM25 mediates IRAK1 K63 polyubiquitination to exaggerate IFNI responses in an RIG-I-dependent manner. We have also observed LDLR+ macrophages displaying heightened IFNI responses in BCG-treated human macrophages. To antagonize LDLR degradation by PCSK9 inhibitors increases IFNI responses in the macrophages and accelerated viral clearance. Our study thus couples LDLR upregulation to antiviral activity in BCG-trained macrophages, making commercial PCSK9 inhibitors potential antiviral indications in clinic.

Staphylococcus aureus is an opportunistic pathogen that poses a considerable burden to healthcare settings worldwide, aided by its ability to thrive in different environmental growth conditions and survive exposure to antibiotics. Small regulatory RNAs (sRNAs) are decisive in enhancing bacterial fitness by modulating gene expression in response to changing environmental conditions. We investigated the role of sRNAs in the adaptation of S. aureus to antibiotics. By assessing the fitness of a library of sRNA mutants, we identified that RsaA sRNA is required for optimal bacterial growth when exposed to low concentrations of fluoroquinolone, a class of antibiotics targeting DNA replication. We also found that in the absence of RsaA, S. aureus is less susceptible to β-lactam antibiotics, which act on the cell wall. RsaA has been reported to prevent the expression of MgrA, a master regulatory protein controlling the expression of efflux pumps. Here, we show that RsaA affects the sensitivity of S. aureus to fluoroquinolone and β-lactam antibiotics through MgrA. RsaA has two forms, a short one commonly referred to in RsaA studies, and a long form about twice the length, of which less is known. Interestingly, our phenotype was only restored when complemented with the long form of the gene or when it was supplied in two parts, the short form and the missing part to obtain the long form. This work demonstrates the role of regulatory RNAs in the adaptation of S. aureus to antibiotic resistance and highlights their value as potential therapeutic targets for manipulating individual sRNA responses to promote the efficacy of existing antibiotics.

Although cisplatin is an effective anticancer agent for treating ovarian cancer, it encounters significant resistance. A full understanding of the mechanisms behind cisplatin resistance has not been achieved. This study identifies MBNL2 as a crucial regulator of cellular responses to cisplatin, examining variations in gene expression and methylation profiles between cisplatinsensitive and -resistant ovarian cancer cells. Cells resistant to cisplatin exhibited increased MBNL2 mRNA expression and significant demethylation at promoter CpG sites. Treating ovarian cancer cell lines with a DNA demethylating agent significantly raised MBNL2 mRNA expression, indicating that epigenetic mechanisms involving DNA methylation control MBNL2 expression. Modulating MBNL2 levels altered the response to cisplatin through survival pathways that shield cells from cisplatin-induced apoptosis. Overexpressing MBNL2 enhanced resistance, while its depletion heightened cisplatin sensitivity. Furthermore, MBNL2 mRNA levels differed among patients based on their response to platinum-based chemotherapeutics. Patients resistant to these drugs had higher MBNL2 mRNA levels, effectively distinguishing them from those who were sensitive (AUC = 0.89, P = 0.0308). A meta-analysis of seventeen datasets confirmed that lower MBNL2 expression levels are associated with a better chemotherapy response and longer relapse-free survival. Conversely, higher MBNL2 expression levels correlated with increased recurrence rates and reduced survival. Thus, MBNL2 may serve as a promising prognostic and therapeutic target for overcoming cisplatin resistance. [BMB Reports 2025; 58(5): 224-231].

TNBC is an aggressive metastatic cancer that poses considerable treatment challenges because of its acquired drug resistance towards the existing targeted and hormonal therapies. The epigenetic modulation including HDACs triggers the EMT in TNBC which produces a more aggressive tumor phenotype. Chemotherapy and radiotherapy cause severe side effects which make treatment complex and challenging. To avoid these serious side effects and boost the effectiveness of current anti-cancer medications, plant flavonoids have been investigated.

Metformin is an antidiabetic drug used in type 2 diabetes as well as indicators in polycystic ovary syndrome (PCOS) and cancer. Due to their increase in popularity, high amounts of metformin are being released into aquatic environments. However, the toxic effect of metformin on embryonic development in aquatic organisms remains limited. Therefore, this study aimed to elucidate the lethal embryotoxicity of metformin and determine the underlying molecular pathways influencing embryonic development using a zebrafish model through multi-omics analysis. Metformin was microinjected into zebrafish embryos at the 1-cell stage with varying concentrations (50 mM, 100 mM, 200 mM, 400 mM, and 800 mM). From the results, hatching rates decreased in a dose dependent manner. Fetal malformation and mortality (LC50 = 339.8 mM) increased in a dose dependent manner. In situ hybridization of whole-embryo assays demonstrated that metformin exerts a significant impact on the initial stages of embryonic development, leading to aberrant differentiation of the germ layers, perturbed organogenesis, and delayed development. Furthermore, transcriptomics, metabolomics, and lipidomics were used to study the molecular mechanisms of embryonic toxicity. The results showed that the cell cycle, dorsoventral axis formation, and collecting duct acid secretion pathways were significantly altered in treated embryos. In brief, these results provide useful information on the lethal toxicity mechanism of metformin overdose and provide clues for further studies in humans.

Endocrine-disrupting chemicals (EDCs) in the aquatic environment are an emerging concern and can lead to adverse health effects on humans and aquatic life. EDCsare ubiquitous in several daily use and personal care products and ubiquitous in aquatic ecosystems. The aquatic ecosystems also serve as major sinks of EDCs and have even been found to accumulate in aquatic organisms. Fish are an important sentinel species in the aquatic system and are a reliable indication of environmental water pollution. In the present study, we have assessed the immunotoxicity effects of three important EDCs, i.e., triclosan (TCS), bisphenol A (BPA), and diethyl phthalate (DEP). There is mounting evidence that EDCs impact several physiological systems, including fish immune systems. Hence, to better understand the immune system's complexity, we have investigated how EDCs alter the immune responses and can aggravate immunotoxicity using Labeo catla as a model fish species. The results showed significant upregulation of immune gene expression; exposure to EDCs differentially modulates immunity across the different organs (liver and brain) of Labeo catla. The present study highlighted that endocrine-disrupting compounds (TCS, BPA, and DEP) have a significant immunotoxicity effect in fish and activate several immunological pathways to control the toxic effect and maintain homeostasis. The results also indicate that immune genes can be used as a biomarker for EDC toxicity. However, further studies need to see how immune-disrupting effects happen at actual exposure levels in the environment to EDCs.