Staphylococcus aureus is a major human pathogen mainly involved in chronic biofilm-related infections, especially at epithelial surfaces and wound sites, owing to its ability to form antibiotic-resistant biofilms and release virulence factors. Conventional antibiotics often fail to eradicate established biofilms and may contribute to the emergence of drug resistance, underscoring the urgent need for alternative biofilm-inhibiting and antivirulence strategies. Herein, we synthesized several dicyclic peptides, including cyclo(L-Phe, L-Hyp) and its stereoisomers cyclo(L-Phe, D-Hyp) and cyclo(D-Phe, L-Hyp), to evaluate their ability to inhibit S. aureus biofilm formation. All cyclic dipeptides exhibited a minimum inhibitory concentration (MIC) of 1 mg/mL. Biofilm inhibition was assessed via crystal violet staining and confocal laser scanning microscopy. At 0.5 mg/mL (1/2 MIC), the compounds exhibited superior inhibition of 24 h biofilm formation compared with vancomycin, with cyclo(L-Phe, D-Hyp) showing the most profound inhibitory activity. qRT-PCR revealed that at 0.25 mg/mL (1/4 MIC), cyclo(L-Phe, D-Hyp) significantly downregulated the expression of the Agr-quorum-sensing system (RNAIII: -75.7%; hla: -61.7%; psm-α: -73.3%), the ica operon (icaA: -71%; icaD: -76.7%), and sarA (-69.3%) (P < 0.0001). Notably, cyclo(L-Phe, D-Hyp) showed low cytotoxicity (CC50 = 5.13 ± 0.27 mg/mL) and negligible hemolysis (<1 %) at twice its MIC, indicating a favorable safety margin for antimicrobial use. These cyclo dipeptides can be formulated for topical delivery to sites such as skin, mucosa, or open wounds, providing a practical approach for localized treatment of biofilm-associated infections. These findings identify cyclo(L-Phe, D-Hyp) as a promising lead for the development of topical anti-infective agents targeting chronic S. aureus biofilm-associated infections.

Avicennia marina, a pioneer mangrove species, has adapted to the intertidal habitat along the tropical and subtropical coasts by developing salt glands on its leaf epidermis. Jasmonic acid (JA) is known to regulate the development of various plant epidermis. However, its role in the development of salt glands in A. marina remains unclear. In this study, we treated A. marina seedling using exogenous methyl jasmonate (MeJA) to investigate the effect of JA on the development and cell fate determination of salt glands, stomata and trichomes in A. marina leaf. The results showed MeJA significantly increased both the density of salt glands and the Na⁺ secretion. Besides, MeJA treatment positively regulated the trichome initiation and negatively affected stomatal lineage ground cells, with a significant decrease in stomatal density but no significant change in trichome density, while it exhibited that salt gland cells may partially originate from trichomes or stomatal lineage cells. Moreover, qRT-PCR results indicated that MeJA affects salt gland development via influencing the process of cell cycle, like reducing endoreduplication. These findings clarify how salt glands contribute to A. marina adaptation to coastal intertidal habitat from a tissue development perspective.

Renal cell carcinoma (RCC) is a highly prevalent malignant tumor of the urinary system, and resistance to the targeted drug sunitinib remains a major clinical challenge. This study investigated the mechanism underlying the action of Astragalus polysaccharide (APS) in RCC and demonstrated that APS can degrade ZEB1 via the ubiquitin proteasome pathway, downregulate the expression of downstream SCD1, and inhibit the Wnt/β-catenin signaling pathway. In vitro experiments revealed that the combination of APS and sunitinib significantly suppressed the proliferation, migration, and invasion of renal cancer cell lines such as 786-O and A498. Additionally, this combination promoted apoptosis and induced G1 phase cell cycle arrest. In vivo validation in a nude mouse xenograft model confirmed that this combined regimen synergistically reduced the tumor volume without causing obvious organ toxicity. Clinical data further verified that high ZEB1 expression is associated with poor prognosis in RCC and that ZEB1 promotes tumor progression by regulating SCD1 to activate the Wnt signaling pathway. In conclusion, APS enhances the sensitivity of RCC to sunitinib by targeting the ZEB1-SCD1-Wnt axis, thus providing a theoretical basis for the clinical application of this combined therapy.

Metformin may exert anticancer effects in non-small cell lung cancer (NSCLC) by influencing long non-coding RNAs involved in the disease's pathology. The Study aimed to investigate the inhibitory effect of metformin on NSCLC by modulating the expression of lncRP11-242D8.1.

Osteoclasts and foreign body giant cells (FBGCs) are multinucleated cells derived from monocytes, but they have distinct functions. Osteoclasts resorb bone while FBGCs form in response to foreign material. Regarding bone implants, osteoclasts are responsible for implant integration, but also for bone resorption associated to implant loosening, while FBGCs play a role in the immune response to the foreign material. Little is known about which proteins in the local environment fine-tune the multinucleation of osteoclasts or FBGCs. One candidate is Activin A (ActA). It has been shown to induce larger, more active osteoclasts, but its effect on FBGC differentiation is unknown. We investigated the effect of ActA on the differentiation of osteoclasts and FBGCs from human CD14-positive monocytes. The number of multinucleated cells and the cell area was measured. qPCR was performed to assess the effect of ActA on gene expression. ActA's influence on osteoclast and FBGC formation was studied on plastic, bone and hydroxyapatite coated Titanium discs (ALD-HA). ActA induced fewer, but bigger and more active osteoclasts on plastic and bone. In contrast, ActA did not have an effect on FBGC number. On ALD-HA, ActA reduced the number of FBGCs, but did not influence osteoclast numbers. qPCR showed that ActA upregulated the expression of several genes such as TRAcP, CIITA, OLR1, RHOBTB1 and ALK4, but mainly in osteoclasts. These results show that ActA has a different effect on osteoclasts compared to FBGCs. This difference could be caused by a difference in the expression in the canonical ActA receptor ALK4.

Ovulation is induced via a surge in gonadotrophin hormones, which increases the expression of the essential ovulatory transcription factor progesterone receptor (PGR) and its target genes. The importance of PGR in ovulation is well defined; however, the role of its many downstream genes largely remains unknown. Using mouse models of ovulation, we show that the Epas1 gene, which encodes the hypoxia inducible transcription factor 2α (HIF-2α), is expressed in a PGR-dependent manner during ovulation. Numerous HIF target genes increase in expression upon gonadotrophin stimulation in mouse granulosa cells, with expression of Epas1, but not its related isoform, Hif1a, increasing in a PGR-dependent manner. PGR directly binds introns of the Epas1 locus to enhance chromatin accessibility in ovarian granulosa cells in vivo, yet no evidence of PGR-dependent Epas1 expression was observed in PGR-expressing breast cancer cell lines, suggesting ovary-specific mechanisms of PGR-dependent Epas1 regulation. PGR activation in response to hormonal stimulation induced expression of a HIF reporter system in primary human granulosa cells, with HIF-2 inhibition with the small molecule PT-2385 confirming a HIF-2 contribution to this response. Upon HIF-2 inhibition with PT-2385 in mice, no change in ovulation counts were observed. However, gonadotrophin-induced ovary gene expression was significantly disrupted, supporting a model where HIF-2α contributes to the control of periovulatory gene expression downstream of PGR. In particular, inflammatory gene expression was dysregulated and a cohort of gonadotrophin-dependent genes, including Pgr, were elevated, suggesting impaired downregulation post-ovulation. These findings provide an important insight into regulation of the hypoxia inducible transcription factors during ovulation and how targeting HIF-2α may be of benefit in future fertility treatments.

Flocoumafen (FCF), a second-generation anticoagulant rodenticide (SGAR), has limited toxicological data regarding its effects on aquatic organisms. In this study, we exposed zebrafish embryos to FCF solutions at concentrations of 0.2, 0.4, and 0.8 mg/L until 120 h post-fertilization (hpf). The results revealed a decrease in survival rates. Notably, FCF exposure significantly reduced the frequency of spontaneous tail coils at 24 hpf, while shortened body length and induced spinal curvature at 120 hpf. Furthermore, zebrafish larvae exhibited craniofacial abnormalities and incomplete bone mineralization at 120 hpf following FCF exposure. In Tg(HuC:EGFP) transgenic strains, neuronal loss was observed. Additionally, FCF-exposed zebrafish larvae showed a marked reduction in locomotor ability, activity levels, and turning capacity. The qPCR and enzyme activity assays revealed significant changes in gene expression associated with the Notch signaling pathway, accompanied by increased levels of reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA). Astaxanthin (ASTA) partially alleviated the toxicities induced by FCF. These findings suggest that FCF may induce skeletal and neurological toxicities by affecting oxidative stress, disrupting the normal expression of skeletal and nervous system-related genes in the Notch signaling pathway, and ultimately leading to behavioral abnormalities. Our findings may provide new insights into a comprehensive evaluation of FCF toxicology in aquatic organisms, and may assist the government in formulating and implementing regulatory policies regarding the application of FCF.

Wnt/β-catenin signaling pathway is the most frequently altered cascade in colorectal cancer (CRC), and the nucleus accumulation of β-catenin is the hallmark of the activation of Wnt/β-catenin signaling. However, despite extensive research efforts, the molecular regulators governing β-catenin nuclear translocation remain poorly understood. The deubiqutinase enzyme USP22 was involved closely in the tumorigenesis of CRC with the underlying mechanism remaining unclear. In our study, we identified USP22 was upregulated in CRC and was closely associated with the activation of Wnt/β-catenin signaling. In the mechanism study we demonstrated for the first time that USP22 deubiquitinated β-catenin in a deubiquitinase activity-dependent manner to facilitate its nuclear translocation and enhance Wnt pathway activation, thereby promoting CRC cell proliferation. Furthermore, we revealed USP22 auto-deubiquitylated and stabilized itself. Additionally, we identified Berberine (BBR), a potent USP22 inhibitor, effectively suppressed Wnt pathway activation and CRC cell proliferation by targeting USP22. Notably, BBR reduced tumor stemness, exhibited substantial toxicity towards cancer stem cells and overcame the chemoresistance. Together, our finding elucidated USP22 promoted CRC progression via deubiquitinating β-catenin and promoting its subsequent nucleus translocation, and highlighted BBR was a potential therapeutic agent for CRC treatment as a novel USP22 inhibitor.

The transcriptional alterations underlying epithelial-to-mesenchymal transition (EMT) in chemoresistant ovarian cancer remain a matter of debate, with emerging evidence pointing to tumour cell plasticity and subclone reprogramming. In this study, we developed a cisplatin-tolerant ovarian cancer model by treating the cisplatin-sensitive OVCAR-3 cell line with a single dose of cisplatin, generating the OVCAR-3 CP variant. These cisplatin-tolerant cells exhibited distinct EMT-related changes at both transcriptomic and protein levels, potentially regulated by epigenetic mechanisms. EMT profiling revealed that OVCAR-3 CP cells did not display a pronounced mesenchymal phenotype but rather retained epithelial characteristics and showed elevated expression of ALDH3A1. In contrast, the parental chemosensitive OVCAR-3 cells expressed canonical mesenchymal markers (CDH2, VIM, ZEB1/2, SNAIL, SLUG) and lacked stemness marker expression. Upon xenografting, both OVCAR-3 and OVCAR-3 CP cells demonstrated phenotypic plasticity, with parental OVCAR-3 xenografts acquiring EMT-like features resembling to those observed in cisplatin-tolerant tumours. These findings suggest a decoupling of EMT from cisplatin-tolerance and instead underscore a stronger association between stemness traits and cisplatin tolerance. Our data further indicate that xenografting can induce significant cellular reprogramming. Comprehensive characterization of ovarian cancer cell-derived xenograft is therefore essential, as they represent a valuable translational platform for investigating therapy adapted ovarian cancer cells.

Chemotherapy resistance is the primary cause of clinical treatment failure and unfavorable prognosis among breast cancer patients. Consequently, the exploration of novel molecular targets for chemotherapy resistance is warranted. Here, we demonstrated that Zinc Finger Protein 184 (ZNF184) facilitates chemoresistance in breast cancer. Through integrated bioinformatics and experimental validation, we identified that ZNF184 was highly expressed in paclitaxel-resistant breast cancer cells. Knockdown of ZNF184 inhibited cell proliferation and re-sensitized resistant cells to paclitaxel in vitro and in patients-derived organoids (PDOs). Mechanistically, ZNF184 regulates the expression of stemness-related genes CD44, OCT4, Nanog, SOX2, and ALDH1A1, thereby promoting the proliferation of breast cancer cells and subsequent paclitaxel resistance. Pan-cancer analysis revealed the potential of ZNF184 as a prognostic and predictive biomarker for adverse clinical outcomes. Collectively, these findings reveal a previously unknown role of ZNF184 in breast cancer progression and paclitaxel resistance, providing new insights into ZNF184 as a potential therapeutic target for cancer patients.

Non-small cell lung cancer (NSCLC) is a metabolism associated disease which mainly depends on anaerobic glycolysis to produce the macromolecules needed for biosynthesis and rapid cell proliferation. Since the cancer cells depend on glycolysis for energy production, the development of drug targets that inhibit the glyco-metabolism will be a promising approach for the management of NSCLC. Plant derived phytocompounds have demonstrated anti-NSCLC activity by modulating the glycolytic pathway, thus curbing the energy requirement essential for the proliferation of cancer cells. In the current study, we explored the efficacy of farnesol in A549 lung adenocarcinoma cells using in vitro assays. Farnesol inhibited the viability of A549 cells to 50% at 21.5 µg/mL. Relative proteomic profiling via nano LC-MS/MS analysis identified 277 differentially expressed proteins in control and farnesol treated samples. Notably, PKM (FC = -3.911819), TKT (FC = -2.857373), ALDOA (FC = -4.8557) and LDH (FC = -2.624372) were downregulated exhibiting a strong interacting network in STRING analysis indicating suppression of anaerobic glycolysis. Furthermore, a decrease in the expression of GluIIβ, FBKP1A and apoptotic regulators such as LAP2 and ATP5F subunits suggest initiation of autophagy and apoptosis. AO/EtBr staining confirmed a late apoptotic shift while, DAPI staining revealed nuclear fragmentation at this concentration. Additionally, farnesol impaired mitochondrial ATP synthesis by reducing mitochondrial membrane potential (MMP) to 66% and elevated ROS levels to 54% creating a disturbance in mitochondrial stability. Overall, Farnesol significantly disrupts anaerobic glycolysis in A549 cells promoting cell death through mitochondrial dysfunction, oxidative stress, apoptosis and reducing cellular acidosis.

Oxaliplatin resistance significantly impairs therapeutic outcomes in colorectal cancer. However, reliable diagnostic markers for early detection of resistance remain limited. This study aimed to identify novel diagnostic signatures through integrative bioinformatics and machine learning approaches.

To investigate the effects of fungal volatile organic compounds (FVOCs) on the mycelial growth of Ganoderma lucidum, and to elucidate the molecular mechanisms underlying the growth-promoting effect of 3-octanone. G. lucidum was cultivated with 1-octen-3-ol, 3-octanol and 3-octanone for 7 days, after which colony diameter and mycelial dry weight were measured to assess their effects on mycelial growth. RNA-seq was used to investigate gene expression changes following 3-octanone exposure. While 1-octen-3-ol or 3-octanol inhibited mycelial growth in G. lucidum, 3-octanone promoted it. In total, 590 differentially expressed genes (DEGs), including 162 upregulated and 428 downregulated genes, were identified following 3-octanone exposure. Functional annotation revealed that among the DEGs, 23 genes were related to fungal cell wall biosynthesis and remodeling, whereas 21 genes were involved in plant-derived polysaccharide degradation. Furthermore, significant expression changes were observed in genes related to secondary metabolism. Our results indicate that G. lucidum can use 3-octanone as a signal to recognize other fungi, potentially facilitating the expansion of its own territory within wood in nature.

Neuroblastoma (NB) is the most common extracranial solid malignancy of children, and MYCN amplification defines a high-risk subtype with poor outcomes. Although widely used in preclinical drug discovery, NB cell lines are often selected based on availability rather than the molecular characteristics of patient-derived tumors, leading to a critical translational gap between experimental outcomes and clinical relevance. To address this, we developed a rank-based transcriptomic correlation framework to assess the concordance between patient-derived tumors (n = 642; combined from the SEQC/MAQC-III and TARGET cohorts) and publicly available NB cell lines (n = 39). This system-level analysis enabled the identification of cell model representatives (CMRs) that closely recapitulate the gene expression landscapes of clinical tumors. COG-N-557, SMS-KAN, and NB-SD emerged as the top CMRs for MYCN-amplified tumors, whereas COG-N-549, FELIX, and SK-N-SH were identified for MYCN-nonamplified tumors. Pathway enrichment analyses indicated that MYCN-amplified CMRs retain key transcriptional programs involved in neuronal development and tumor proliferation, supporting their biological relevance. Leveraging these models, we integrated pharmacogenomic connectivity mapping and drug-gene network analyses to uncover kinase inhibitors and epigenetic modulators as promising therapeutic candidates capable of targeting MYCN-driven transcriptional programs, despite MYCN being an undruggable oncogene. In conclusion, this study addresses a fundamental systems biology and translational research gap by establishing a data-driven framework for selecting NB cell lines that accurately reflect patient-derived tumor biology with direct implications for prioritizing therapeutically relevant drug candidates. Future studies should prioritize the top CMRs as in vitro models to enhance translational relevance and accelerate precision drug discovery in high-risk pediatric NB.

Acute pancreatitis (AP) remains a challenging clinical condition with limited therapeutic options and high mortality rates in severe cases. Traditional anti-inflammatory approaches have shown disappointing results in clinical trials, highlighting the urgent need for novel therapeutic strategies targeting the underlying pathophysiological mechanisms. The study by Jia et al presents compelling evidence for a previously unrecognized mechanism through which rutaecarpine, a bioactive alkaloid from traditional Chinese medicine, exerts protective effects against AP. This research demonstrates that rutaecarpine alleviates AP by targeting the epigenetic machinery, specifically through enhancer of EZH2-mediated suppression of FBXW11. The authors employed both in vitro cerulein-induced AR42J cell models and in vivo sodium taurocholate-induced rat models to establish the therapeutic efficacy of rutaecarpine and elucidate its molecular mechanisms. Their findings reveal that rutaecarpine upregulates EZH2 expression, leading to increased histone H3 methylation at the FBXW11 promoter region, thereby suppressing FBXW11 expression and consequently reducing inflammatory infiltration and oxidative stress. The significance of this work extends beyond demonstrating rutaecarpine's protective effects. It identifies FBXW11 as a novel therapeutic target in AP and provides the first evidence that traditional Chinese medicine compounds can modulate epigenetic reprogramming in pancreatic inflammation. Recent studies have confirmed FBXW11's role as an inflammatory biomarker in pancreatitis, supporting the clinical relevance of this pathway. The study's comprehensive approach, combining molecular docking, cellular thermal shift assays, and co-immunoprecipitation studies, strengthens the mechanistic insights. These findings open new avenues for AP treatment by targeting epigenetic regulators rather than relying solely on conventional anti-inflammatory strategies, potentially leading to more effective therapeutic interventions for this devastating condition.

Parkinson's disease (PD) is characterized by mitochondrial dysfunction and impaired protein homeostasis, with the mitochondrial unfolded protein response (mtUPR) emerging as a key regulatory pathway in mitigating mitochondrial stress. This study aimed to explore the impact of shRNAs targeting CHCHD2 or FBXO7 on the mitochondrial unfolded protein response (mtUPR) in a Parkinson's disease (PD) cell model, clarify the mitochondrial-nuclear signaling pathways involving CHCHD2 and FBXO7, elucidate the mechanisms underlying mitochondrial dysfunction induced by these genes, and identify new therapeutic targets for early stage PD. An in vitro PD model was established by treating SH-SY5Y cells with MPP+; mitochondrial morphology was evaluated using transmission electron microscopy, and qRT-PCR and Western blot were employed to determine the expression levels of mRNAs and proteins associated with mtUPR, autophagy, CHCHD2, and FBXO7 under oxidative stress. In the MPP+-induced PD cell model, we knocked down CHCHD2 and FBXO7 via shRNA and treated the cells with JNK and AKT agonists to observe their effects on mtUPR protein expression. The results showed that mtUPR was activated in MPP+-exposed SH-SY5Y cells, and the expression of CHCHD2 and FBXO7 genes was significantly upregulated after MPP+ intervention; knockdown of CHCHD2 via shRNA resulted in a marked decrease in the expression of mtUPR-related proteins such as HSPA9, HSPD1, YME1L1, and CLPP, while shRNA targeting FBXO7 exerted only a minimal effect on these mtUPR proteins. Furthermore, the administration of JNK or AKT agonists significantly enhanced the expression of MPP+-induced mtUPR proteins, including HSPA9, HSPD1, YME1L1, and CLPP. Collectively, these findings indicate that CHCHD2, rather than FBXO7, plays an essential role in modulating the MPP+-induced mtUPR and suggest that CHCHD2 may regulate mitochondrial protein homeostasis by activating the mtUPR through the JNK/c-Jun and AKT/ERα pathways.

Polygonum cognatum (Madımak) is a plant traditionally consumed for medicinal purposes in Turkey. Unlike previous studies examining samples from different regions and seasons, this research presents the first comprehensive characterization of P. cognatum collected from the Central Black Sea Region (Tokat, 40°01'02″N, 36°28'15″E; 1210 m altitude) during the vegetative growth phase (June 2024), where geographical origin and collection time significantly influence secondary metabolite profiles. This study evaluates the phytochemical profile and multitarget biological activities of P. cognatum extracts obtained using solvents of different polarities (hexane, ethanol, and water). Advanced analytical techniques (liquid chromatography-tandem mass spectrometry, high-performance liquid chromatography-diode array detector, and gas chromatography-mass spectrometry) identified 28 phenolic compounds, with the ethanol extract showing the highest diversity (24 compounds) and total phenolic content (78.6 ± 2.3 mg GAE/g). Compounds identified for the first time in P. cognatum include isoquercetin-3-O-rhamnoside, apigenin-7-O-glucoside, and luteolin-4'-O-glucoside. The ethanol extract demonstrated superior multitarget bioactivity: potent antioxidant activity ( 2,2-diphenyl-1-picrylhydrazyl (DPPH) IC50: 76.4 ± 2.1 μg/mL), moderate but selective anti-inflammatory effects (COX-2 IC50: 145.3 ± 5.2 μg/mL; selectivity index: 2.06, indicating preferential COX-2 inhibition over COX-1) and significant antidiabetic potential (α-amylase IC50: 89.3 ± 3.1 μg/mL; α-glucosidase IC50: 76.8 ± 2.9 μg/mL), and antimicrobial activity (MIC: 62.5 μg/mL against S. aureus). Notably, this study demonstrates for the first time the histone deacetylase (HDAC) inhibitory activity of P. cognatum (IC50: 92.4 ± 3.8 μg/mL), revealing novel epigenetic modulation properties. Molecular docking studies showed strong correlations between binding affinities and experimental IC50 values (r = -0.87 to -0.91; p < 0.01). Cytotoxicity evaluation showed favorable safety profiles (CC50 > 500 μg/mL). Docking, IC50, and compositional data consistently indicate that quercetin, rutin, chlorogenic acid, and kaempferol are key contributors to the observed antioxidant, antidiabetic, anti-inflammatory, and HDAC inhibitory effects. These findings establish P. cognatum as a promising multitarget therapeutic agent with novel epigenetic regulatory mechanisms, supporting its potential development for inflammatory, metabolic, and epigenetic-related disorders.

Epigenetic memory enables the propagation of gene expression patterns following transient stimuli. Although three-dimensional chromatin organization is emerging as a key regulator of genome function, it is unknown whether it contributes to cellular memory. Here we establish that acute perturbation of the epigenome can induce cellular memory of gene expression in mouse embryonic stem cells. We uncover how a pulse of histone deacetylase inhibition translates to changes in transcription, histone modifications and genome folding. While most epigenomic and transcriptional changes are initially reversed once the perturbation is removed, some loci remain transcriptionally deregulated and genome architecture partially maintains its perturbed conformation. Consequently, a second pulse of transient hyperacetylation induces stronger memory of transcriptional deregulation. Using ultradeep Micro-C, we associate memory of gene expression with repressive Polycomb-mediated chromatin topology. These results demonstrate how cells can record transient stresses in their genome architecture, thereby enabling an enhanced response to subsequent perturbations.

This study investigates the role of arachidonate 5-lipoxygenase (ALOX5) in upregulating programmed death-ligand 1 (PD-L1) expression via its metabolite 5-hydroxyeicosatetraenoic acid (5-HETE) in ovarian cancer cells, and explores the effects on lipid metabolism and ferroptosis. Additionally, the study examines the role of Tectorigenin (TEC) in this process. Using network pharmacology, bioinformatics analysis, molecular docking, and drug affinity reaction target stability (DARTS) experiments, we identified ALOX5 as a potential therapeutic target for TEC in ovarian cancer treatment, possibly through lipid metabolism and ferroptosis pathways. In vitro experiments showed that TEC inhibits proliferation and invasion and promotes apoptosis in ovarian cancer cells without significant cytotoxicity in normal ovarian epithelial cells. TEC also inhibits lipid metabolism and promotes ferroptosis, reducing ovarian cancer cell viability. Inhibition of ALOX5 similarly suppresses lipid metabolism and enhances ferroptosis, effects that can be reversed by exogenous 5-HETE. In vivo studies using a nude mouse model demonstrated that TEC inhibits tumor growth and downregulates ALOX5, 5-HETE, and PD-L1 expression in tumor tissues. The findings suggest that the ALOX5/5-HETE signaling pathway is crucial for regulating lipid metabolism and ferroptosis in ovarian cancer, and TEC may exert its anti-tumor effects, at least in part, by modulating this pathway (The graphical abstract was shown in Fig. 1).

Cancer signaling encompasses a wide array of entangled molecular cascades that promote oncogenic progression and counteract the effect of tumor suppressors. Transforming growth factor β (TGFβ) induces complex and stage-dependent effects throughout tumor progression. During pre-malignant hyperplastic growth, TGFβ restricts cell proliferation and inflammation, while on the other hand, TGFβ promotes migration and distal metastasis of cancer cells. To dissect the temporal chromatin-based transcriptional response to TGFβ, we employed 3D culture models of isogenic human breast epithelial cells, exemplified by non-oncogenic MCF-10A (MI) and their HRAS-transformed counterpart (MII). Genome-wide chromatin accessibility profiling revealed an extensive chromatin opening induced by TGFβ at transcription start sites and enhancer elements in both models, with a marked enrichment of SOX4 binding motifs in oncogenic cells. Transcriptomic analyses unexpectedly revealed the upregulation of DNA replication and DNA damage response pathways, following TGFβ stimulation of oncogenic MII 3D cultures. Canonical TGFβ-driven programs, including epithelial-mesenchymal transition and metabolic reprogramming, were activated in both models. Notably, single-cell RNA-seq of primary breast tumors confirmed co-expression of SOX4 and cell cycle regulators. Mechanistically, we show that TGFβ induces the interaction between the MH2 domain of SMAD3 and the intrinsically disordered regions of SOX4, co-activating downstream gene targets. Validating the genome-wide analyses, we found that resistance of breast cancer cells to the CDK4/6 inhibitor palbociclib conferred by TGFβ stimulation was functionally dependent on SOX4. Collectively, our findings reveal an apparent oncogenic function of TGFβ in promoting cell cycle progression and drug resistance through SOX4, highlighting the pro-tumorigenic role of TGFβ signaling in breast cancer progression.