The unfolded protein response (UPR) is a highly conserved adaptive mechanism that restores endoplasmic reticulum (ER) homeostasis under stress. Beyond its canonical roles in proteostasis, the UPR has emerged as a central regulator of immune responses across diverse contexts, including infection, inflammation, cancer, and autoimmunity. IRE1α, PERK, and ATF6 are three principal UPR sensors that coordinate complex signaling networks to regulate antigen presentation, cytokine production, and immune cell differentiation. This review highlights the molecular mechanisms by which small molecules target the UPR to modulate immune responses. In addition, we highlight stress granules (SGs) and the prevalence of protein-protein interactions mediated by intrinsically low-complexity domains (LCDs) in the UPR as potential new avenues for immune modulation. Finally, we discuss future directions for leveraging UPR modulation in immunotherapy, infectious disease, and chronic inflammatory disorders.

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, with resistance to 5-fluorouracil (5-FU) representing a major therapeutic challenge. Thymoquinone (TQ), a bioactive constituent of Nigella sativa, exhibits anticancer activity; however, the mechanisms underlying TQ-induced cell death appear to be highly context dependent. This study aimed to characterize cell line-specific death pathways triggered by TQ in colorectal cancer models with distinct molecular backgrounds and differential responsiveness to 5-FU. Human CRC cell lines RKO (5-FU-sensitive) and SW1116 (poorly responsive), along with normal colon epithelial cells (CCD-841CoN), were treated with TQ, 5-FU, or their combination for 24 h. Cell viability, DNA fragmentation, caspase-3/7, -8, and -9 activity, cell death phenotypes, and expression of apoptosis- and necroptosis-related genes were evaluated using MTT assays, ELISA, luminescent assays, flow cytometry, and RT-qPCR. TQ significantly reduced viability in both CRC cell lines while exerting minimal cytotoxicity toward normal cells. In RKO cells, characterized by microsatellite instability (MSI), TQ induced DNA fragmentation, caspase activation, and transcriptional upregulation of pro-apoptotic genes, consistent with apoptosis-associated signaling. In contrast, SW1116 cells, which exhibit chromosomal instability (CIN) and reduced responsiveness to 5-FU, displayed decreased viability accompanied by suppressed caspase activity and predominant features of caspase-independent necrotic cell death. This differential response may be attributed to the CIN phenotype, which has been associated with impaired apoptotic signaling and enhanced tolerance to cytotoxic stress. Combined TQ and 5-FU treatment did not produce synergistic cytotoxicity, as confirmed by Bliss independence analysis, but revealed distinct, cell line-dependent death programs. These findings demonstrate that TQ modulates cell death execution in a molecular context-dependent manner rather than enhancing 5-FU efficacy through pharmacological synergy.

Background:Malva sylvestris (the common mallow) is an herbaceous species widely used in ethnobotanical practices to treat gastrointestinal, hepatic and urinary inflammation. Objectives: Despite these beneficial effects on human health, the antineoplastic potential of this plant has not yet been fully explored. Thus, in the present study, two human colon cancer cell lines (i.e., HCT-116 and Caco-2) were treated with an extract obtained from M. sylvestris flowers (MFE), whose composition in terms of phytochemicals and microRNAs has been recently published by our research group, to explore its potential bioactivity. Methods/Results: MTT and Trypan blue assays demonstrated that MFE reduced tumour cell growth without causing significant cytotoxicity or apoptosis. Following the diphenylboric acid 2-aminoethyl ester-induced fluorescence of some plant metabolites, microscopy analysis proved that MFE components crossed the cell membranes, accumulating into nuclei. Wound assay and transwell tests documented that MFE was also able to reduce cell motility and invasiveness. In both cell lines qPCR experiments demonstrated that MFE caused the over-expression of factors, like VIMENTIN and E-CADHERIN, which negatively influence epithelial-mesenchymal transition in colon cancers. However, the effects of MFE appeared to be time-, dose- and cell type-dependent. In fact, the treatment induced senescence in P53-null Caco-2 cells (i.e., ROS, β-galactosidase and P21WAF1/Cip1) and a premise of differentiation (i.e., P27Kip1) in P53-wild-type HCT-116 cells, also via the CDK2/c-MYC/AKT axis, justifying its antiproliferative property. In parallel, the transfection of tumour cells with pure synthetic miR160b-5p-a microRNA identified in M. sylvestris flowers and predicted to target the human CDK2 transcript-resulted in gene silencing, thereby suggesting its central role in mediating the cross-kingdom effects of MFE on the investigated cancer models. Conclusions: Overall, these findings open new perspectives on the common mallow as a source of potential antimetastatic compounds and on the possible use of its plant microRNAs in the development of gene therapies.

Cancer remains a leading global cause of death, with conventional treatments often limited by toxicity and recurrence. Recent advances in gene therapy and nanodrug delivery offer new avenues for precision oncology. Human telomerase reverse transcriptase (hTERT) and glutathione peroxidase 4 (GPX4) are overexpressed in many cancers and linked to apoptosis and ferroptosis, respectively. Here, we developed a manganese dioxide nanosheet (MnO2-NS) probe co-loaded with antisense oligonucleotides targeting hTERT and GPX4 mRNA to synergistically down-regulate both genes and induce dual cell death pathways. The probe, assembled via adsorption of fluorescently labeled antisense strands, showed controllable release in the presence of glutathione (GSH). Cellular uptake and antisense release were confirmed in multiple cancer cell lines. The MnO2-NS probe significantly suppressed cell proliferation, outperforming single-target or carrier-only controls. Molecular analyses confirmed reduced hTERT and GPX4 expression, along with GSH depletion, ROS accumulation, and elevated lipid peroxidation-collectively promoting enhanced cancer cell death. In summary, this MnO2-NS-based co-delivery system enables synergistic gene silencing and GSH depletion, enhancing antitumor efficacy and providing a promising strategy for multifunctional nanotherapy.

We investigated the protective effects of skipjack tuna (Katsuwonus pelamis) bone-derived biocalcium (Bio) against dexamethasone-induced atrophy in C2C12 myotubes. Bio rescued atrophic morphology, increasing myotube diameter dose-dependently. It mitigated inflammation by suppressing nitric oxide production and the expression and concentration of proinflammatory cytokines (IL-6, TNF-α, IL-1β) significantly and dose-dependently. Bio restored protein turnover balance by downregulating MuRF1 and atrogin-1 while upregulating MTOR. At 5-20 µg/mL, Bio downregulated total NF-κB p65, p38 mitogen-activated protein kinase (MAPK), and FoxO3a and upregulated Akt expression. Crucially, Bio dose-dependently downregulated primary-, precursor-, and mature-microRNA-29b. In Bio-treated, dexamethasone-treated C2C12 myotubes, microRNA-29b inhibitor co-transfection significantly increased myogenin and MyoD expression, whereas microRNA-29b mimic co-transfection suppressed these myogenic markers, confirming the inhibitory role of microRNA-29b. Molecular docking simulations confirmed strong binding affinities between microRNA-29b and myogenin/MyoD. These results demonstrate that Bio exerts anti-atrophy effects by disrupting the microRNA-29b-mediated block on myogenesis and modulating inflammatory responses, protein turnover, and key signaling pathways. Collectively, skipjack tuna-derived Bio shows promise as a functional food supplement for sarcopenia prevention and management.

Flatfish metamorphosis is an abrupt post-embryonic transformation driven by thyroid hormones (THs), in which a bilaterally symmetric pelagic larvae becomes an asymmetric benthic juvenile. While the craniofacial changes associated with eye migration during metamorphosis are well documented, the role of THs in central nervous system (CNS) remodelling remains poorly understood. Here we investigated the role of THs on CNS remodelling during metamorphosis of the flatfish, Solea senegalensis, by integrating high-throughput transcriptomic analysis with experimental manipulation of TH availability using an inhibitor of hormone synthesis, methimazole (MMI) or exogenous T4. Transcriptome profiling revealed 567 differentially expressed gene transcripts associated with TH-levels involved in CNS development, neuronal and glial differentiation, migration, myelination and metabolism. Key CNS-related factors such as klf9, sox9, mbp, and plp were strongly down-regulated in MMI-treated larvae. Cell proliferation assays further demonstrated increased interocular neural proliferation under hypothyroidism, consistent with impaired differentiation. Region-specific analyses of the head and body uncovered distinct patterns of TH signalling involving dio2, dio3, thra, thrb, and mct8, underscoring the spatial complexity of endocrine regulation. These results highlight that THs are crucial for both morphological remodelling and CNS plasticity during flatfish metamorphosis, underscoring their conserved role in vertebrate brain development.

Acute myeloid leukemia (AML) with mutated nucleophosmin 1 (NPM1) presents therapeutic challenge and exhibits a striking female predominance in clinical. However, the contribution of gender-specific factors, such as sex hormones to the pathogenesis and therapeutic implications in NPM1-mutated AML remains elusive. In the present study, we found that NPM1-mutated AML showed female bias, particularly among the young, pre-menopausal age (20-49 years) group, and that female exhibited inferior overall survival compared to males. Subsequently, 17β-estradiol (E2), the primary circulating estrogen in pre-menopausal females, was shown to facilitate the proliferation of NPM1-mutated leukemia cells. Importantly, ovariectomy (OVX) alleviated the leukemia burden, while reintroduction of exogenous E2 accelerated disease progression in leukemia mouse models. Mechanistically, leukemia cells exhibited an estrogen receptor α (ERα)-dominant/ERβ-low expression signature, which was maintained through NPM1 mutant-induced ubiquitination-dependent degradation of ERβ. The diminished ERβ level allowed ERα to act as the primary responder of E2, evidenced by the nuclear translocation of ERα following E2 treatment. Transcriptomic analysis identified hepatocyte growth factor (HGF) as the potential target of E2 signaling, and E2-mediated ERα activation transcriptionally upregulated HGF, thus promoting leukemia cell proliferation. Therapeutically, blockade of the E2/ER pathway using selective ER degrader fulvestrant inhibited the growth of NPM1-mutated AML cells, and its combination with chemotherapy drug cytarabine induced synergistic anti-leukemia effects in vitro and in vivo. Collectively, these data demonstrated the oncogenic role of E2/ERα signaling in NPM1-mutated leukemia and suggested that targeting E2/ERα signaling combined with standard chemotherapy may represent a promising regimen for female patients.

TetR family transcriptional regulators (TFRs) represent a major class of regulatory proteins that govern diverse cellular processes, including antibiotic biosynthesis, primary and secondary metabolism, and stress responses. In this study, the role of a specific TFR, TetR1, in organic solvent tolerance of Rhodococcus ruber SD3 (R. ruber SD3) was investigated through functional characterization. The interaction between TetR1 and its target DNA was confirmed via molecular docking and electrophoretic mobility shift assay. Under various organic solvent stresses, the relative expression level of the tetr1 gene was downregulated in R. ruber SD3, indicating that it acted as a negative regulator of organic solvent tolerance. Comparative growth analysis showed that the tetr1 knockdown strain exhibited enhanced growth compared to the wild-type under organic solvent stress, whereas the overexpression strain displayed impaired growth within 72 h. Transcriptomic profiling identified 804 differentially expressed genes between the tetr1 knockdown and wild-type strains, including 328 upregulated and 476 downregulated genes. Transcriptomic analysis further revealed that tetR1-KD triggered global transcriptional reprogramming. This included: upregulating ABC transporters while suppressing MCE-family importers to limit solvent influx; remodeling membrane lipid composition by altering fatty acid metabolism gene expression to enhance stability; and coordinately boosting xenobiotic biodegradation pathways to improve detoxification. These results provided new insights into the biological functions of TetR1 in R. ruber.

Prenatal arsenic exposure has been linked to a myriad of negative health effects. There is relatively little insight into the mechanisms and signaling alterations across different fetal organs that drive long-term immune-related issues following prenatal arsenic exposure. Therefore, the effects of this exposure window on gene expression in the liver, placenta, heart, and lung of gestation day (GD) 18 C57BL/6 mouse fetuses were investigated. From two weeks prior to mating until tissue collection at GD18, mice were exposed to 0 or 100 ppb sodium (meta) arsenite in drinking water, ad libitum. Genes of interest were analyzed by RT-qPCR, complemented with untargeted Agilent 44 K microarray analysis. Data cleanup and analysis was performed in RStudio. Differentially expressed mRNAs were queried in the String Database and using Cytoscape to create interaction networks and identify significantly enriched biological pathways. A total of 251, 165, 158, and 41 genes were significantly altered in the liver, placenta, heart, and lung, respectively, when treated samples were compared to controls. Many altered pathways were immune-related, supporting prior research. Most notably, gene expression of Gbp3, a key player in the cellular response to interferon gamma, was found to be reduced in placentas of male fetuses exposed to arsenic compared to controls (p = 0.0237). IMPACT: This is the first study comparing alterations in gene expression across multiple organs following prenatal exposure to environmentally relevant levels of arsenic. These findings, elucidating the multi-organ impact of prenatal arsenic exposure on predominantly immune-related pathways, further our mechanistic understanding of the long-term health effects observed in early-life arsenic-exposed populations.

Sarcomas are a heterogeneous group of cancers with few shared therapeutic targets. We show that PI3K signaling is frequently activated in sarcomas due to PTEN loss (in 30%-60%), representing a common therapeutic target. The PI3K pathway has lacked a downstream oncogenic transcription factor. We show TAZ and YAP are transcriptional coactivators regulated by PI3K and drive a transcriptome necessary for tumor growth in a PI3K-driven sarcoma mouse model. This PI3K/TAZ/YAP axis exists in parallel to the known PI3K/AKT/mTORC1 axis, providing a rationale for combination therapy targeting the TAZ/YAP-TEAD interaction and mTORC1. Combination therapy using IK-930 (TEAD inhibitor) and everolimus (mTORC1 inhibitor) synergistically diminished proliferation and anchorage-independent growth of PI3K-activated sarcoma cell lines at low, physiologically achievable doses. Furthermore, this combination therapy showed a synergistic effect in vivo, suggesting that an integrated view of PI3K and Hippo signaling can be leveraged therapeutically in PI3K-activated sarcomas.

The paradigm of targeted therapy has been revolutionized by the development of proximity-inducing chimeras, particularly proteolysis-targeting chimeras (PROTACs), which offer a therapeutic strategy for previously "undruggable" targets. However, the degradation of certain essential proteins can lead to off-target toxicity, prompting the development of nondegradative proximity-inducing chimeras. This review categorizes current nondegradative chimeras into several functional classes, including targeted inhibition, targeted stabilization, post-translational modification (PTM), and cellular switches; elaborates on the mechanisms of action for each category; and introduces representative technologies, including molecular glues (MGs). Furthermore, the review highlights cutting-edge research and recent advancements in the field. Unlike previous reviews focusing primarily on degradative chimera technologies, this article systematically categorizes nondegradative modalities into inhibition, stabilization, PTM, and cellular switches, providing a unified mechanistic framework and highlighting emerging therapeutic applications beyond oncology.

Irinotecan, a standard therapeutic agent for metastatic colorectal cancer (mCRC), often faces significant limitations due to drug resistance, with treatment failure observed in approximately 30%-50% of patients, leading to poor clinical outcomes. This study aims to systematically elucidate the molecular mechanisms underlying irinotecan resistance in colorectal cancer (CRC) by constructing patient-derived organoid (PDO) models combined with single-cell transcriptomics technology.

Parental exposure to toxicants can affect progeny health. However, laboratory studies often employ exposures that result in loading of pollutants to gametes or toxic effects to parents, which could indirectly affect germ cell or gamete health. Here, we took advantage of the biology of Caenorhabditis elegans to carry out a study in which we minimized the potential for maternal loading of toxicants, and used an exposure paradigm that either did (high concentration) or did not (low concentration) significantly impact the health of the P0 generation. We hypothesized that parental exposure to mitochondrial toxicants during germ cell and gamete development, at levels not causing P0 toxicity, would result in altered mitochondria and organismal health in offspring. In the P0 generation, a high rotenone concentration altered growth, mitochondrial respiration, gene expression, induction of the mitochondrial unfolded protein response, and susceptibility to dopaminergic neurodegeneration induced by a chemical rechallenge later in life. However, we observed minor or no effects in P0 at a low concentration. In high-exposure F1 offspring, we observed altered embryo size, larval developmental stage distribution, spare respiratory capacity, heat shock protein expression, and dopaminergic neurodegeneration after a secondary rotenone challenge. The only effects observed in the F1 offspring of the low exposure were a 1.7% decrease in egg size (size later in development was normal), and moderate evidence of a slightly increased sensitivity to heat shock protein expression and dopaminergic neurodegeneration caused by a secondary later-in-life rotenone exposure. We recommend that parental toxicity be carefully assessed to contextualize offspring outcomes.

Bovine mastitis is a prevalent and economically devastating disease in the global dairy industry. Antibiotic overuse leads to increased antimicrobial resistance and reduced milk quality, becoming major bottlenecks in clinical treatment. Transfer factor (TF), a safe, low-cost, and readily available immunomodulator that enhances cell-mediated immunity, has emerged as a promising antibiotic alternative. This study aimed to investigate TF's alleviative effect on bovine mastitis and its underlying molecular mechanisms. We found that clinical mastitis cows had significantly higher mRNA levels of interleukin-1β (IL-1β) and interleukin-6 (IL-6) and markedly lower expression of tight junction (TJ) genes (nectin cell adhesion molecule 4 [NECTIN4], tight junction protein 1 [ZO-1], occludin) in mammary tissue and milk somatic cells compared to healthy controls. In vitro experiments, TF pretreatment inhibited the secretion of pro-inflammatory cytokines (IL-1β and IL-6) and the expression of TJ genes and proteins (NECTIN4, occludin, ZO-1) in a graded manner across the tested concentrations (up to 180 µg/mL) in lipopolysaccharide (LPS)-induced bovine mammary epithelial cells (MAC-T). Mechanistically, TF alleviated TJ protein inhibition by regulating myosin light chain kinase (MLCK) activity, mimicking the effect of MLCK inhibitor ML-7. It also mitigated LPS-induced changes via inhibiting the nuclear factor κB (NF-κB) pathway, similar to NF-κB inhibitor Bay 11-7082, and blocked NF-κB activation by inhibiting the transforming growth factor-β-activated kinase 1 (TAK1) pathway, comparable to TAK1 inhibitor Takinib. In vivo, intramammary infusion of TF via the teat canal into the infected quarter of cows with subclinical mastitis downregulated IL-1β/IL-6 mRNA, upregulated TJ genes, and reduced both MLCK expression and the phosphorylation of myosin light chain (MLC), p65, and TAK1. By day 5, TF group's (n = 17) California mastitis test (CMT) negative conversion rate reached 64.7% (vs. 13.3% in the normal saline (NS) group, n = 15) with a significantly lower somatic cell count (SCC). Our findings demonstrate that TF, by concurrently eliciting anti-inflammatory and barrier-repair effects via inhibition of the TAK1/NF-κB/MLCK axis, effectively alleviates bovine mastitis. This study furnishes a robust molecular framework for deploying TF as a non-antibiotic tool in the sustainable control of mastitis.

Interest in the utilization of novel plant-based extracts for supporting gut health and nutrition in companion animals is growing. The effect of betaine on canine intestinal barrier integrity and immune-related gene and protein expression by epithelial and macrophage-like cells was evaluated under conditions of inflammatory challenge from lipopolysaccharide (LPS). We evaluated: (1) the effect of betaine pre-treatment (0 to 1,000 µg/mL for 24 h) on the viability of canine intestinal epithelial MCA-B1 and macrophage-like DH82 cells; (2) epithelial barrier function of LPS-challenged MCA-B1 cells pre-treated with betaine, by measurement of 4-kDa fluorescein isothiocyanate (FITC-dextran [FD4] flux); (3) gene and protein expression of tight junction proteins and adhesion molecules using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and proteomics; (4) anti- and pro-inflammatory and regulatory gene expression in LPS-challenged, betaine pre-treated, MCA-B1, DH82 and peripheral blood mononuclear cells and; (5) the phagocytic activity of DH82 cells pre-treated with betaine. No cytotoxicity of betaine against MCA-B1 or DH82 cells were observed. Betaine pre-treatment of MCA-B1 reduced FD4 permeability compared with untreated cells (P < 0.05), accompanied by increased (mRNA) expression of claudin-1 (P < 0.001) and E-cadherin (P < 0.05) and increased protein expression of junctional adhesion molecules (P < 0.05), vinculin (P < 0.05) and aquaporin-3 (AQP3; P < 0.01). Betaine pre-treatment of LPS-challenged cells also increased the expression of regulatory cytokines TGF-β1 (P < 0.01), TGF-β2 (P < 0.01), and IL-33 (P < 0.05) in MCA-B1 cells and reduced the expression of IL-6 (P < 0.05) and MyD88 (P < 0.01) in DH82 cells compared with untreated cells. In addition, betaine enhanced the phagocytic activity of DH82 cells (P < 0.01) compared with untreated cells. Confirmatory studies in vivo are needed but these results suggest that plant-derived betaine has potential as a nutritional supplement for companion animals to support gut health and immunity.

Heterozygous mutations in isocitrate dehydrogenase (IDH) 1 and 2 are hallmarks of astrocytoma, IDH-mutated, and oligodendroglioma, IDH-mutated, as defined by the World Health Organization Classification of Tumors of the Central Nervous System, 5th Edition. Mutant IDH confers a neomorphic enzymatic activity that converts α-ketoglutarate (α-KG) into the oncometabolite D-2-hydroxyglutarate (D-2-HG), which inhibits α-KG-dependent dioxygenases and induces a global DNA hypermethylation phenotype, also known as Glioma CpG Island Methylator Phenotype (G-CIMP). To elucidate mechanisms underlying the antitumor effects of DS-1001b-a novel, brain-penetrant, orally available inhibitor of mutant IDH1 R132H and R132C-we performed preclinical analyses using IDH1 R132H-mutant glioma cells in vitro and orthotopic mouse xenograft models (MGG152, BT142, and A1074). DS-1001b treatment reduced 2-HG levels in vitro and in vivo and significantly prolonged survival in A1074 and BT142 intracranial xenograft models (p = 0.0064 and 0.0004, respectively), confirming effective target inhibition in the brain. In vitro, prolonged DS-1001b exposure partly reversed genome-wide DNA hypermethylation and revealed that H3K4me3 modulation was mostly associated with differential gene expression, affecting pathways related to apoptosis, necrosis, cell cycle arrest, and migration in MGG152. Metabolomic analyses further demonstrated a significant reduction in asparagine in A1074, consistent with the activation of L-asparaginase-mediated pathways. Collectively, these findings indicate that sustained DS-1001b administration exerts antitumor effects in IDH1-mutant glioma mouse models and induces transcriptomic, epigenetic, and metabolic reprogramming.

Glioblastoma (GB) is one of the most aggressive malignant brain tumors. Due to the high invasiveness of this cancer, surgical removal is often not possible, and relapses after surgery are very common, making current treatments ineffective. Developing new therapies or treatment combinations remains a major challenge in managing GB. Metformin (MET), an anti-diabetic medication, has recently gained attention for its potential anticancer effects. To better understand how MET inhibits GB growth at the molecular level, we studied its impact on survivin, a member of the inhibitor of apoptosis (IAP) family that is essential for GB cell survival, resistance to radio- and chemotherapy, and tumor recurrence. Using T98G and U87-MG cell lines, we performed cell viability, migration, and invasion assays, along with Western blot analysis, ChIP assays, and gene silencing experiments to examine key signaling pathways. We found that MET effectively inhibits the growth, viability, and invasiveness of GB cell lines through a molecular mechanism involving activation of the AMPK/FoxO3a/survivin pathway. In vivo studies support these findings, showing increased FoxO3a and decreased survivin in brain tissue sections from metformin-treated mice compared with untreated controls. These results suggest new possibilities for repurposing MET as an adjuvant treatment for GB.

N6-methyladenosine (m6A) has emerged as a pivotal regulator of post-transcriptional gene control, yet its contribution to chemotherapy resistance remains insufficiently defined. Here, we describe a previously unrecognized METTL3-ADAM23 epitranscriptomic regulatory relationship associated with platinum (Pt) resistance in ovarian cancer (OC). We show that cisplatin treatment increases global m6A levels and METTL3 expression, linking Pt exposure to activation of the m6A machinery. Functional perturbation studies demonstrate that METTL3 overexpression enhances cisplatin resistance, whereas METTL3 knockdown or pharmacologic inhibition with the selective METTL3 inhibitor STM2457 sensitizes OC cells to Pt treatment in vitro and improves Pt response in vivo. Transcriptomic profiling identifies ADAM23, a cell-adhesion-related tumor suppressor, as a METTL3-dependent, m6A-associated transcript, with altered mRNA expression observed across multiple experimental systems and several high-confidence predicted m6A sites within its transcript. Cisplatin-associated METTL3 upregulation correlates with reduced ADAM23 expression, suggesting a potential regulatory relationship that may contribute to chemoresistance. Together, these findings support a model in which METTL3-associated increases in m6A methylation are linked to Pt resistance, in part through modulation of ADAM23 expression, and highlight METTL3 as a potential therapeutic target in OC.

SATB2 (special AT-rich binding protein 2) functions as a chromatin-associated epigenetic regulator that modulates gene expression, in part by serving as a transcriptional cofactor. This study assessed whether SATB2 overexpression is sufficient to promote in vitro transformation of human mesothelial cells and whether SATB2 suppression in mesothelioma cancer stem cell (CSC)-enriched populations is associated with altered chemoresistance. SATB2 expression was high in human malignant pleural mesothelioma (MPM) cell lines but absent in Met5A mesothelial cells. Ectopic SATB2 expression in Met5A cells was associated with acquisition of malignant and stem cell-like phenotypes, including increased expression of stem cell markers and pluripotency-associated factors, as well as anchorage-independent growth in soft agar and spheroid formation in suspension culture. In contrast, Met5A cells transduced with an empty vector did not form colonies or mesospheres. SATB2 overexpression in Met5A cells was also associated with increased motility, migration, and invasion, accompanied by induction of epithelial-mesenchymal transition (EMT)-related transcription factors relative to empty vector controls. Conversely, shRNA-mediated SATB2 knockdown in an MPM cell line attenuated proliferation, EMT-associated features, and CSC-like characteristics. Chromatin immunoprecipitation assays identified SATB2 occupancy at promoter regions of Bcl2, XIAP, KLF4, c-Myc, NANOG, and SOX2, consistent with a role in transcriptional regulation of genes linked to transformation, pluripotency, cell survival, proliferation, and EMT. In CSC-enriched cells, SATB2 inhibition was associated with increased sensitivity to cisplatin and pemetrexed, concomitant with reduced OCT4 and SOX2 expression. Collectively, these findings support SATB2 as a candidate therapeutic target in MPM and suggest that SATB2 suppression may enhance chemotherapy response when combined with standard agents.

Ovarian cancer remains one of the leading causes of cancer-related mortality among women, underscoring the need for novel combination strategies that effectively inhibit tumor cell growth while limiting adverse effects. N-acetylcysteine (NAC) and alpha-ketoglutarate (AKG) are biologically active compounds with reported anticancer properties; however, their combined effects in ovarian cancer are not well characterized. In this study, we applied an integrative approach combining network pharmacology analysis with in vitro experiments to investigate the effects of NAC and AKG on OVCAR3 ovarian cancer cells. Common molecular targets of NAC and AKG were identified by intersecting predicted compound targets with ovarian cancer-associated genes, followed by protein-protein interaction network construction and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses. Experimental validation assessed the effects of NAC and AKG, alone and in combination, on cell viability, apoptosis, migration, and clonogenic capacity. Network analysis identified 70 shared target genes enriched in pathways related to apoptosis, cellular stress responses, and cell migration. In vitro experiments demonstrated that combined treatment with NAC (10 mM) and AKG (100 µM) significantly reduced cell viability, increased apoptotic cell death, and markedly suppressed cell migration and colony formation compared with single-agent treatments. Overall, these findings indicate that the combination of NAC and AKG exerts enhanced inhibitory effects on ovarian cancer cell growth and motility in vitro.