Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy that has a poor survival rate of ∼13 % with limited options for effective therapies. DDX3 is a member of the DEAD-box RNA helicase enzyme family. It acts as an adapter protein that interacts with several transcription factors, enhancing their binding ability to the promoters of genes involved in cancer progression. Previously, we demonstrated that PAF1, a component of the RNA polymerase II-associated factor 1 complex, interacts with DDX3 to promote PDAC stemness. Here, we investigated the therapeutic efficacy of RK-33, a small-molecule inhibitor targeting DDX3, in combination with gemcitabine (GEM) and 5-fluorouracil (5FU), which enhanced therapeutic efficacy in KRAS-driven PDAC.-DDX3 and PAF1 exhibit progressively increased expression in various stages and correlate well with poor survival of PDAC. Targeting DDX3/PAF1 significantly mitigated clonogenic, EMT, and stemness phenotypes in PDAC cells. It also reduced tumor growth, proliferation, and increased apoptosis in xenograft and PDAC organoid models. Finally, MXRA5, EDIL3, COL13A1, and SLC16A2 were identified as top downstream response genes upon RK-33 treatment and potential new targets to mitigate extracellular matrix remodeling, angiogenesis, cell migration, and cell cycle progression, thereby enhancing the therapeutic efficacy of GEM and 5FU Overall, our data indicate that RK-33 enhances the therapeutic efficacy of GEM and 5FU in mitigating PDAC aggressiveness. Consequently, these findings open new avenues for developing efficacious therapeutic adjuvants to treat advanced pancreatic cancer.

Biliverdin reductase A (BLVRA) is a key enzyme in bilirubin metabolism, where it reduces biliverdin to bilirubin. Bilirubin is a potent antioxidant that protects cells from oxidative stress. Therefore, reduced or deregulated BLVRA activity may contribute to increased oxidative DNA damage, which is one of the factors leading to the neoplastic transformation of cells.

G-quadruplex structures within the promoter regions of some oncogenes diminish transcriptional activity. The suppression and downregulation of Cyclooxygenase-2 (COX-2), which is encoded by the prostaglandin-endoperoxide synthase 2 (PTGS2) gene, could control the size of colorectal cancer tumors. This study aimed to investigate the impact of a mononuclear octahedral cobalt(III) Schiff base complex [CoL3] (L = 2-((allylimino)methyl)-6-methoxyphenol) on the G-quadruplex structures in the PTGS2 promoter region and to assess its downregulation effects in the human colorectal cancer cell line HT-29. At first, molecular docking was used to evaluate the binding of [CoL3] to the PTGS2 promoter region. Then, the potential and stabilization of G-quadruplex formation in the PTGS2 promoter motifs were investigated using circular dichroism (CD) spectroscopy, polyacrylamide gel electrophoresis, and polymerase chain reaction (PCR) stop assays. COX-2 expression was also assessed by real-time PCR and western blot. Molecular docking analysis indicates that [CoL3] has a potent interaction with the PTGS2 promoter region. Additionally, [CoL3] has an intense inhibitory effect on the PTGS2-PCR stop assay. The CD measurements confirmed the conformational property of G-quadruplex DNA structures induced by [CoL3]. Treatment of colorectal cancer cells with the [CoL3] resulted in a modest increase in PTGS2 expression by a mean factor of 1.69 (p < 0.05). Conversely, the expression of COX-2 proteins was downregulated significantly, with a mean protein intensity of 0.47 (p < 0.001). Therefore, [CoL3] induces G-quadruplex formation in the promoter region of PTGS2, which ultimately inhibits protein expression of COX-2. This means that [CoL3] can be an effective agent for colorectal cancer treatment.

Timosaponin AIII (Tim-AIII), a steroidal saponin derived from Anemarrhena asphodeloides, has emerged as a promising antitumor agent, yet its precise molecular targets and mechanisms in breast cancer remain poorly defined. Here, we identify fibroblast growth factor 2 (FGF2) as a direct binding target of Tim-AIII using a combination of network pharmacology, CETSA, and surface plasmon resonance assays. Mechanistically, Tim-AIII exhibits a dual therapeutic mode of action. First, it induces reactive oxygen species (ROS)-mediated endoplasmic reticulum (ER) stress, activating the eIF2α-ATF4-CHOP axis and initiating apoptosis. Second, it dampens the FGF2-FGFR1-PI3K/AKT signaling cascade, thereby inhibiting epithelial-mesenchymal transition (EMT) and suppressing cell migration and invasion. RNA sequencing and enrichment analyses confirm that Tim-AIII regulates critical oncogenic pathways, including ER stress, calcium signaling, and PI3K/AKT. In vivo evaluations demonstrate that Tim-AIII significantly reduces tumor growth without detectable systemic toxicity in breast cancer-bearing mice. This study not only elucidates the molecular basis of Tim-AIII's antitumor efficacy but also positions it as a potential targeted therapeutic for breast cancer, with dual action on ERS-induced apoptosis and EMT suppression.

Osteoporosis is a common skeletal metabolic disorder. Cardamonin (CAR) is a natural chalcone compound with multiple activities. However, the role and mechanism of CAR in osteoporosis progression remain largely unknown.

Although immunotherapy has reshaped the treatment landscape for gastrointestinal (GI) malignancies, durable clinical benefit is achieved in only a subset of patients, largely due to the emergence of immune resistance. Accumulating evidence highlights histone lactylation, an epigenetic modification driven by excessive lactate accumulation, as a pivotal mediator linking tumor metabolism to immune suppression. By reprogramming transcriptional networks within the tumor microenvironment, histone lactylation skews macrophages toward an immunosuppressive phenotype and compromises T-cell effector function, thereby facilitating tumor immune escape. In metabolically active GI tumors with pronounced lactate production, aberrant histone lactylation contributes to the establishment of a profoundly immunosuppressive niche that undermines responses to immune checkpoint inhibitors. Targeting lactylation-related pathways has therefore gained attention as a novel therapeutic avenue, with the potential to restore antitumor immune activity, enhance cytotoxic lymphocyte function, and sensitize tumors to immunotherapy. This article summarizes current understanding of the crosstalk between lactate metabolism, histone lactylation, and immune regulation, and highlights therapeutic approaches targeting this epigenetic axis to enhance immunotherapy efficacy in GI cancers.

Triple-negative breast cancer (TNBC) is distinguished by high invasiveness and a tendency for recurrence. Recent studies have suggested that E3 ubiquitin ligases play a crucial role in the initiation and progression of various tumors. However, there is still an absence of systematic understanding regarding the specific function and molecular mechanisms of its member gene RNF130 in TNBC.

Alcohol is a major cause of hepatocellular carcinoma (HCC), accounting for 30% of cases worldwide. Sorafenib, a tyrosine kinase inhibitor (TKI), was the standard first-line treatment for advanced HCC until 2021, but sorafenib resistance is common. We explored the impact of chronic alcohol exposure (CAE) on sorafenib response and sought to identify associated resistance mechanisms. Huh-7 HCC cells were chronically exposed to alcohol for 6 months. Sorafenib resistance was assessed by measuring cell viability (IC50) and by evaluating the protein expression of signaling pathways involved in resistance using immunoblotting. RNA sequencing was performed to identify mechanisms of resistance. Sorafenib response was assessed using the RECIST 1.1 criteria in HCC patients. A retrospective study of 86 HCC patients from the CHIEF cohort (alcohol-related vs. non-alcohol-related etiologies) evaluated overall survival (OS) and progression-free survival (PFS) using the log-rank test. CAE significantly decreased cell sensitivity to sorafenib (p = 0.006), indicating increased resistance. The ERK pathway was involved. RNA sequencing of our cells identified a total of 80 differentially expressed genes associated with drug resistance and aggressiveness. Clinically, alcohol-related HCC patients were less responsive to sorafenib (35% responders vs. 65%, p = 0.014) and had significantly different OS (p = 0.0234). Median OS was 10 months (95% CI = [6.1, 15.7]) for alcohol-related HCC and 12.1 months (95% CI = [7.7, 64.9]) for other etiologies. PFS was lower in the alcohol group (5.72 months (95% CI = [4.63, 12.8]) vs. 9.66 months (95% CI = [4.40, 39.9], p = 0.0298). Sorafenib resistance due to chronic alcohol consumption is consistent in both in vitro models and clinical settings. KEY MESSAGES: Chronic alcohol exposure reduces the effectiveness of sorafenib in hepatocellular carcinoma (HCC), as demonstrated in both in vitro and clinical settings. In vitro, alcohol-exposed HCC cells showed increased sorafenib resistance, associated with activation of the ERK signaling pathway and differential expression of 80 genes linked to drug resistance and tumor aggressiveness. Clinically, patients with alcohol-related HCC had poorer responses to sorafenib and shorter overall and progression-free survival compared to patients with non-alcohol-related HCC. These findings suggest that alcohol-related HCC may require alternative or personalized therapeutic strategies beyond standard TKI treatments.

Uveal melanoma (UM) is among the most prevalent intraocular malignant tumors worldwide. Celastrol exhibits broad-spectrum anticancer properties; however, its underlying therapeutic mechanism in UM is yet to be elucidated. In this study, a network pharmacology approach was employed to identify potential common targets of celastrol and UM. These targets were further analyzed in conjunction with transcriptomic data and machine learning algorithms, which led to the identification of CTNNB1 and STAT3 as key molecular targets. The functional roles of these targets were investigated through immune infiltration analysis and single-cell RNA sequencing (scRNA-seq), while the binding stability between celastrol and CTNNB1/STAT3 was assessed using molecular docking (MD) and molecular dynamics simulation (MDS). Subsequently, celastrol was administered to B16-F10 and C918 cell lines, demonstrating that it significantly suppresses cell proliferation and migration by downregulating CTNNB1 and STAT3 expression, while simultaneously inducing apoptosis and cell cycle arrest. Moreover, real-time quantitative PCR (qPCR) and western blot (WB) analyses corroborated the modulation of target expression levels. Therefore, celastrol exerts potent anti-tumor effects in UM by inhibiting the CTNNB1 and STAT3 signaling pathways, thereby suppressing tumor cell proliferation and metastasis, as well as promoting cell cycle arrest and apoptosis.

Bufei Formula (BFF), a traditional Chinese medicine, has been widely utilized in the clinical treatment of chronic obstructive pulmonary disease (COPD), though its underlying mechanisms remain unclear and warrant further investigation.

In melanoma, SOX9 and SOX10 are markers of the mesenchymal and melanocytic state, respectively. Using a panel of BRAFV600E positive YUMM lines, we find that, following chronic vemurafenib treatment, SOX10 is lost whereas SOX9 is induced. Overexpression or knock-down of either SOX9 or SOX10 had no impact on vemurafenib sensitivity. However, we find that SOX9 is necessary to program a vemurafenib-resistance memory state following a drug holiday in vitro. RNA-Seq studies show that the loss of Sox10 represents an intermediate state that is accompanied by the loss of Sox6 and the induction of Sox7, Sox9 and other phenotype switching markers. However, SOX7 expression is not sufficient to induce vemurafenib resistance. Upon acquired drug resistance, we observed differential chromatin accessibility in the Sox9 and Sox10 upstream regions, supporting their activation and repression, respectively. Overall, our data show that the loss of SOX10 and SOX9 induction are critical to program drug resistance. Furthermore, we show that the YUMM cell lines represent a good murine model to investigate transitions to an acquired drug resistant state.

Faecalibacterium species are keystone commensals of the human gut, contributing to intestinal homeostasis, immune modulation, and epithelial health. However, their extreme sensitivity to oxygen and reactive oxygen species renders them highly vulnerable during inflammatory conditions, severely limiting their therapeutic application. Understanding the molecular mechanisms underlying their oxidative stress responses is therefore critical for harnessing these bacteria as next-generation probiotics to restore gut health. In this study, we investigated oxidative stress responses in Faecalibacterium duncaniae A2-165 using comprehensive proteomic and membrane fatty acid profiling. We demonstrated that increasing hydrogen peroxide (H₂O₂) concentrations extend the lag phase of growth and affect survival during the first hour of exposure, notably altering the redox potential. Exposure to H₂O₂ triggered a remodeling of the proteome, including detoxification systems, metal transporters, DNA repair systems, transcriptional regulators, and enzymes involved in cobalamin biosynthesis. Complementary RT-qPCR analyses revealed coordinated and time-dependent transcriptional activation of genes involved in oxidative stress response. Remarkably, cobalamin supplementation enhanced bacterial growth, mitigated H₂O₂-induced stress, and lowered superoxide levels in F. duncaniae, highlighting its direct antioxidant activity. By analyzing membrane fatty acid profiles, we showed that cobalamin preserves membrane fluidity, counteracting oxidative stress induced by H₂O₂ in F. duncaniae. These findings reveal the multifaceted strategies employed by F. duncaniae to withstand oxidative stress and provide a foundation for future efforts to optimize its production at industrial scales and its therapeutic potential as a next-generation probiotic.

Bexarotene is a retinoid X receptor (RXR) agonist that plays a crucial role in cell growth and differentiation. It has shown potential in treating both early- and late-stage cutaneous T-cell lymphoma (CTCL). However, the impact of Bexarotene on the neurodevelopment of aquatic organisms, particularly aquatic vertebrates, remains poorly understood. This study aimed to investigate the effects of various concentrations of bexarotene (3 μg/L, 6 μg/L, and 9 μg/L) on the development of the zebrafish embryonic nervous system, using zebrafish as a model organism. The underlying molecular mechanisms were explored through a combination of pharmacological interventions, molecular biology, histopathology, and transcriptomics. Studies have shown that zebrafish embryos exposed to Bex show significant changes in development, including morphological abnormalities, head malformations, significantly shortened head length and width, reduced fluorescent area, cell apoptosis, shortened spinal motor neuron axon length, abnormal myelin development, decreased oligodendrocytes, cerebellar developmental damage, and abnormal behavior. Transcriptomics and qPCR results showed abnormal expression of neurodevelopmental genes (olig2, mbpa, atoh1a, gfap, ngn1, gap43, etc.). In addition, exposure to medium and high concentrations of bexarotene significantly increased acetylcholinesterase (AChE) activity. Bexarotene activates the Wnt signaling pathway, and treatment with the Wnt inhibitor IWR-1 can partially rescue the neurodevelopmental impairments in embryos. In summary, bexarotene offers new insights into the potential neurodevelopmental risks in zebrafish embryos, emphasizing the importance of preventing drug side effects and ensuring the safe and rational use of medications to protect the health of living organisms.

Nearly half of patients with recurrent pregnancy loss (RPL) link to disrupted maternal immune tolerance. IL-10-production regulatory B cells (Bregs) are functionally impaired in RPL, but the underlying mechanisms remain unclear. This study aimed to identify these mechanisms and test suberoylanilide hydroxamic acid (SAHA) as a potential therapy.

Cystatin B (CSTB) is highly expressed in hepatocellular carcinoma (HCC) tissues and serum, indicating its potential as an early diagnostic biomarker. Given the known tumour-suppressive effects of all-trans retinoic acid (ATRA) in solid tumours, this study investigated whether ATRA inhibits HCC progression by modulating CSTB expression. Bioinformatics analyses of databases revealed the elevated CSTB expression in HCC, correlating with poor patient prognosis. These findings were validated in human HCC tissues and HepG2 cells. Through in vitro and in vivo functional assays, ATRA treatment was shown to significantly inhibit HCC cell proliferation, migration, and invasion, concomitantly with reduced CSTB expression. Proteomic sequencing identified cytochrome b (CYTB), a core component of mitochondrial respiratory chain complex III, as a downstream target of CSTB. Further experiments demonstrated that ATRA decreases mitochondrial membrane potential, complex III activity, and cellular ATP levels, these above effects were partially reversed upon CSTB overexpression. In vivo, ATRA administration effectively suppressed subcutaneous tumour growth. Collectively, these results indicated that ATRA exerts anti-tumour activity in HCC by targeting the CSTB/CYTB axis, thereby impairing mitochondrial function and inhibiting tumour progression.

OCCC has extremely poor prognosis. ARID1A mutation-related chromatin remodeling errors are key molecular features of OCCC. Dysregulation of receptor tyrosine kinases-related signaling pathways is common in OCCC. Here we show that combination of niraparib and lenvatinib exhibits significant synergistic inhibitory effects against platinum-resistant OCCC cell lines, xenografts, patient-derived tumoroid (PDT) models, and prolonged survival in the platinum-refractory patient-derived xenograft (PDX) model. RNA-sequencing revealed the most differently expressed genes in PDX treated with a combination of niraparib and lenvatinib versus control were in structural ribosomal components. Selected differently expressed ribosomal proteins (RPs: RPS2, RPS5, RPS9, and RPL3) was validated using quantitative polymerase chain reaction. Nucleophosmin (NPM1) expression, which is involved in ribosome biogenesis, was inhibited by niraparib and lenvatinib. Our findings imply that combined niraparib and lenvatinib reduces platinum-resistant OCCC progression by attenuating Src phosphorylation, NPM1 expression and ribosome biogenesis. These results highlight the necessity for continued exploration of this promising treatment strategy, particularly in future clinical trials for platinum-resistant or platinum-refractory OCCC. KEY MESSAGES: The combination of niraparib and lenvatinib synergistically inhibits platinum-resistant OCCC. This combination prolongs survival in a platinum-refractory PDX model. RNA sequencing revealed that the most differentially expressed genes in PDX models treated with this combination were associated with structural ribosomal components. This combination suppresses Src phosphorylation, NPM1 expression, and ribosomal protein levels in OCCC.

Irinotecan resistance remains a significant challenge in metastatic colorectal cancer (mCRC) therapy. To address this, we identified USP1 as a synthetic lethal partner of Irinotecan through genome-wide CRISPR/Cas9 screening in HCT-116 cells. Combining the USP1 inhibitor I-138 with Irinotecan in HCT-116, HT-29, and SW620 cell lines significantly reduced IC50, suppressed proliferation, and diminished colony formation compared to monotherapy, demonstrating a synergistic effect (combination index CI < 1). The synergistic therapeutic efficacy was further validated in the xenograft mouse model. Mechanistic studies revealed that I-138 significantly upregulated pCREB (Ser133), concurrently dynamically regulating the activity of USP1, FANCD2/FANCI, and PCNA upon DNA damage response and repair. RNA sequencing further highlighted the enrichment of cAMP, PI3K-AKT, and Wnt pathways, which are all linked to CREB activity in the combination group. These findings establish USP1 inhibition as a promising strategy to overcome Irinotecan resistance through the combination strategy, providing a novel therapeutic avenue for CRC.

Recurrence and metastasis significantly impact the survival outcomes of gastric cancer (GC) patients. Weifuchun (WFC), a well-established traditional Chinese medicine formulation, has been widely used in clinical practice for treating gastric disorders and as an adjunctive therapy following GC surgery.

Thymic stromal lymphopoietin (TSLP) is an epithelial-derived cytokine that induces type 2 immune responses through dendritic cell activation, and its aberrant regulation is implicated in TSLP-associated inflammatory disorders including atopic dermatitis. We previously demonstrated that hypoxia-inducible factor (HIF) suppresses TSLP expression in human keratinocyte cells; however, the underlying mechanism remained unclear. In this study, we aimed to explore the suppressive mechanism of enarodustat, an HIF-prolyl hydroxylase inhibitor. Enarodustat selectively suppressed TSLP expression induced by the fibroblast-stimulating lipopeptide (FSL-1), a toll like receptor 2/6 agonist in HaCaT, a human keratinocyte cell line. Although both the nuclear factor-κB (NF-κB) and activator protein (AP)-1 contributed to FSL-1-induced TSLP induction, enarodustat preferentially attenuated AP-1 signaling by reducing c-Jun N-terminal kinase (JNK) phosphorylation. This JNK dephosphorylation required both HIF1α and HIF2α and was accompanied by increased expression of dual-specificity phosphatases (DUSPs), which target JNK for dephosphorylation. Collectively, our findings identify a previously uncharacterized HIF-DUSP-JNK axis that negatively regulates TSLP expression. This study provides mechanistic insight into how HIF activation shapes epithelial cytokine responses, offering a basis for understanding the pathogenesis of TSLP-associated diseases such as atopic dermatitis.

Noise-induced hearing loss (NIHL) can be either temporary or permanent, depending on the intensity and duration of noise exposure. Excessive noise exposure activates various cellular mechanisms in the cochlea, including oxidative stress, immune responses, and apoptosis. Still, the mechanisms underlying hearing recovery after transient threshold shift (TTS) and lack of recovery after permanent threshold shift (PTS), as well as their therapeutic implications for NIHL, remain unknown. In this study, we performed a comparative analysis of longitudinal changes in the cochlear transcriptome of TTS and PTS mouse models. Our analysis revealed that noise-induced ER stress activates the unfolded protein response (UPR). Notably, the protein kinase R-like endoplasmic reticulum kinase (PERK) branch of the UPR returned to baseline levels following TTS but showed sustained activation following PTS. In addition, the proapoptotic factor C/EBP homologous protein (CHOP) was selectively induced in hair cells following PTS. Administering a PERK inhibitor prior to and following noise exposure hindered hearing restoration after TTS, indicating a requirement for PERK activation in hearing recovery. Inhibition of sustained PERK activation via a PERK inhibitor or reduction of CHOP expression via pharmacological chaperones facilitated partial hearing recovery following PTS. Together, these findings provide insight into the mechanisms underlying NIHL and its prevention, highlighting UPR modulation as a promising therapeutic strategy.