Malignant and non-malignant cells within the tumor microenvironment (TME) actively secrete extracellular vesicles (EVs) that may mediate intercellular communication or enter the blood stream. Circulating EVs in Diffuse Large B-Cell Lymphoma (DLBCL) patients are a promising source of liquid biopsy biomarkers; however, whether different cellular components of the TME preferentially secrete small (S-) and/or large (L-) EVs is still unknown. With an established density-gradient separation protocol and tunable resistive pulse sensing analysis, we demonstrate that DLBCL cells in culture produce 100-1000-fold higher numbers of S-EVs (50-200 nm) compared with L-EVs (200-1000 nm) and very large EVs (>1000 nm). In contrast, the plasma from DLBCL patients contains comparable concentrations of S- and L-EVs, consistent with various cellular origins. Small RNA sequencing showed minor differences in miRNA content between plasma S- and L-EVs; however, messenger RNA sequencing revealed stark differences in cargo between EV-size subtypes and between healthy donors and patients. Deconvolution analysis with single-cell sequencing data from 17 DLBCL tumor tissues as reference using the Statescope algorithm indicated that circulating S-EVs from malignant cells outnumber the L-EVs. In contrast, TME macrophage-, T cell-, and natural killer-derived L-EVs outnumber S-EVs. Together, these findings suggest that circulating S- and L-EVs can originate from distinct cellular compartments within the DLBCL TME, representing complementary biological information. These observations have important implications for the development of EV-based liquid biopsy strategies.

Acute lymphoblastic leukemia (ALL) mainly affects children, though it also occurs in adults. About 25% of adults with ALL have the more aggressive Philadelphia chromosome-positive (Ph+) subtype. Standard treatment includes tyrosine kinase inhibitors (TKIs) in combination with chemotherapy or corticosteroids, which have greatly enhanced survival rates over the past 2 decades. Ponatinib, a third-generation TKI demonstrated greater efficacy compared with imatinib, a first-generation TKI, in the PhALLCON trial.

BackgroundLung cancer remains one of the leading causes of cancer-related mortality worldwide. Holocytochrome c synthase (HCCS), a mitochondrial enzyme involved in apoptosis and energy metabolism, has been implicated in tumorigenesis; however, its role in lung cancer is not well defined.AimThis study aimed to elucidate the prognostic and therapeutic potential of HCCS in lung cancer through integrative bioinformatics analyses.MethodsTranscriptomic, methylation, and clinical data from TCGA were analyzed using TNMplot, UALCAN, and TIMER2.0 to evaluate HCCS expression, promoter methylation, immune infiltration, and prognostic relevance in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). Meta-analysis of 18 independent cohorts from LUNG CANCER EXPLORER and four GEO datasets validated expression patterns, while Kaplan-Meier analysis assessed survival outcomes.ResultsHCCS was significantly upregulated in LUAD and LUSC and showed promoter hypermethylation in LUAD. Meta-analysis and external validation confirmed its overexpression. Patients with low HCCS expression exhibited poorer overall survival, suggesting a potential tumor-suppressive effect. HCCS expression positively correlated with immune cell infiltration and co-expressed genes enriched in mitochondrial and apoptotic pathways.ConclusionHCCS may serve as a prognostic and therapeutic biomarker in lung cancer, linking mitochondrial regulation, epigenetic modification, and immune interactions.

A Japanese phase 2 trial of encorafenib plus binimetinib met the primary endpoint of the centrally assessed objective response rate in patients with unresectable BRAF V600E-mutated thyroid cancer. Consequently, encorafenib plus binimetinib has been approved in Japan. We present the health-related quality of life (HR-QoL) outcomes from the trial.

ObjectiveTo evaluate the efficacy and safety of radiotherapy plus immune checkpoint inhibitors versus radiotherapy plus chemotherapy in driver gene-negative patients with non-small cell lung cancer and brain metastases.MethodsThis single-center retrospective cohort study (Strengthening the Reporting of Observational Studies in Epidemiology-compliant) enrolled 60 consecutive driver gene-negative patients with non-small cell lung cancer and brain metastases (29 radiotherapy plus immune checkpoint inhibitors, 31 radiotherapy plus chemotherapy) treated between June 2018 and December 2023, with follow-up until July 2025. Survival, tumor response, and immune-related adverse events were analyzed using Kaplan-Meier methods, Cox models, and chi-square tests. The study was approved by the Institutional Review Board and used deidentified data.ResultsRadiotherapy plus immune checkpoint inhibitors significantly prolonged median overall survival (586 vs. 509 days, p = 0.0208) and progression-free survival (494 vs. 383 days, p = 0.0127) as well as improved objective response rate (34.48% vs. 19.35%, p = 0.0394) and disease control rate (75.86% vs. 51.61%, p = 0.0265) compared with radiotherapy plus chemotherapy. Favorable prognostic factors included age <60 years, Eastern Cooperative Oncology Group Performance Status <2, programmed death-ligand 1 tumor proportion score ≥50%, and absence of extracranial metastasis. Radiotherapy plus immune checkpoint inhibitors-related immune-related adverse events (24.14%) were mostly grades 1-2, with no grade ≥4 events.ConclusionsRadiotherapy plus immune checkpoint inhibitors may confer survival benefits and favorable safety in driver gene-negative patients with non-small cell lung cancer and brain metastases. However, caution is warranted in interpreting these findings, which require validation in large-scale prospective studies.

Bisphenol A (BPA), an endocrine-disrupting chemical with estrogenic activity, has been implicated in cancer development, although its role remains controversial. This study investigated the effects of BPA on ovarian cancer and its underlying mechanisms. BPA treatment dose-dependently (0-10 μM) increased cell viability and invasion. Kyoto Encyclopedia of Genes and Genomes analysis revealed the enrichment of the central carbon metabolism pathway following BPA exposure. Consistent with this, BPA upregulated glycolytic enzymes HK2 and LDHA. In addition, BPA activated ERα, which enhanced HK2 transcription and promoted glycolysis. The resulting lactate accumulation increased histone H3 lysine 18 lactylation (H3K18la), enriched at the IGF2BP3 promoter, to upregulate its expression. IGF2BP3 then stabilized HK2 mRNA via m6A recognition, amplifying the glycolysis. Our findings suggest that BPA promotes ovarian cancer progression through the HK2/H3K18la/IGF2BP3 sequential regulatory axis, providing insights for epigenetic-targeted therapies.

Precise profiling of multiple circulating tumor DNA (ctDNA) is hindered by the challenge of resolving single-nucleotide variants (SNVs) and their trace abundance. Herein, we present an integrated SERS platform that combines rationally molecularly engineered nanotags with a novel microturbulent electrofluidic interface for multiplexed SNV-level profiling of lung cancer ctDNAs. In particular, the SERS nanotags incorporate (1) a de novo isomeric Raman reporter library that systematically tailors vibrational signatures through molecular symmetry and electronic effects, (2) hollow hyperbranched Cu2O/CuO@Ag heterostructures engineered to suppress carrier recombination for maximized electromagnetic-chemical signal enhancement, and (3) rationally designed dCas9-sgRNA units that confer single-nucleotide discrimination. To overcome diffusion-limited mass transport and further boost the signal, we fabricated nanopebble-structured asymmetric microelectrodes on aluminum foil. Unlike a conventional smooth interface, this unique topography induces powerful microturbulence-enhanced electrohydrodynamics (mt-EHD) that disrupts the interfacial depletion layer to accelerate target delivery while simultaneously offering high-density "hot spots" within interpebble gaps for secondary Raman signal amplification. This integrated system achieved the simultaneous quantification of six ctDNA mutations in 131 clinical samples. By leveraging machine learning algorithms, the platform enables accurate diagnosis, prognostic stratification, and dynamic treatment monitoring, establishing an integrated chemical-electrofluidic strategy for precision liquid biopsy.

Asialoglycoprotein receptor 1 regulates the inflammatory response in granulosa cells associated with polycystic ovary syndrome. In the granulosa cells of individuals with polycystic ovary syndrome, elevated levels of asialoglycoprotein receptor 1 may activate the extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway, which in turn enhances the expression of prostaglandin-endoperoxide synthase 2. Abstract This study evaluated whether asialoglycoprotein receptor 1 (ASGR1) can regulate the expression of prostaglandin-endoperoxide synthase 2 (PTGS2) in granulosa cells (GCs) in polycystic ovary syndrome (PCOS). The expression levels of ASGR1 in GCs and PCOS mice ovaries were assessed; to determine the in vivo role of ASGR1, it was overexpressed in the ovaries of female mice via adeno-associated virus injection, and RNA-sequencing analysis revealed that ASGR1 upregulated prostaglandin-endoperoxide synthase 2 (PTGS2). Western blot confirmed the levels of activated ERK1/2 after ASGR1 overexpression, and ERK1/2 agonists and inhibitors were used to validate the downstream effects on PTGS2. Results showed that the expression level of ASGR1 in GCs and ovaries of women and mice with PCOS was significantly elevated. In vitro cell culture results demonstrated that ASGR1 significantly upregulated the ERK1/2 pathway and increased PTGS2 expression in KGN cells. Moreover, through the use of agonists, small interfering RNA, inhibitors, and overexpression strategies, it was identified that the ERK1/2 signaling pathway mediates the ASGR1-induced elevated expression of PTGS2. It was concluded that the agonist effect of ASGR1 on PTGS2 in GCs in PCOS is mediated by the ERK1/2 pathway, and these findings provide new insights into the pathogenesis of PCOS-associated inflammation.

The Protein Phosphatase Methylesterase 1 (PPME1) is a methylesterase specific to phosphatase 2A, a tumor suppressor, and plays a key role in tumor development. Its impact on pan-cancer diagnosis, prognosis, and immune regulation is still uncertain.

Pancreatic ductal adenocarcinoma (PDAC) continues to pose a significant clinical challenge due to its high mortality rate and limited treatment efficacy. The role of disulfidptosis, a recently discovered mode of regulatory cell death, in pancreatic cancer progression and tumor immunity remains poorly understood.

Originally reported as an oncogene and currently known to be a major "genome guardian", the p53 protein remains one of the most explored transcription factors, exhibiting variety of functions both within transcription regulation and beyond. Given that p53 dysfunction contributes to the majority of human cancers, understanding its regulatory mechanisms and therapeutic potential remains a primary research focus. This review addresses the key aspects of p53 regulation and functionality, analyses its role in tumor evolution, and provides a comprehensive analysis of current and emerging therapeutic strategies targeting the p53, with particular emphasis on immunotherapy approaches.

Breast cancer is one of the leading causes of mortality worldwide. The tumor microenvironment plays a critical role in cancer progression. This microenvironment is composed of various cells embedded in the extracellular matrix (ECM). Laminin-111, a major ECM glycoprotein, produces bioactive peptides that influence tumor biology. We have shown that the laminin-derived peptide C16 (KAFDITYVRLKF), located in the short arm of the γ1 chain, regulates migration, invasion, and invadopodia formation in different cancer cells. Our findings suggest that the regulatory mechanisms underlying the effects of C16 are associated with β1 integrin. This prompted us to investigate the interaction between the C16 peptide and β1 integrin in breast cancer cells. We found that breast cancer cells bind to C16 peptide, and this attachment is inhibited by β1 integrin depletion via siRNA. Cellular localization of the C16 peptide was analyzed using transmission electron microscopy (TEM) and time-lapse fluorescence microscopy. TEM revealed that nanogold-conjugated C16 decorated the cell membrane and was localized in intracellular vesicles, indicating peptide endocytosis. Time-lapse confocal microscopy showed that C16 was internalized by breast cancer cells within 2 h of incubation, with this process increasing over time. Based on these observations, we hypothesized that the peptide is endocytosed and directed to the endosome-lysosome pathway for degradation. Time-lapse imaging demonstrated that part of the internalized peptide colocalized with lysosomes in breast cancer cells. This suggests that C16 may be involved in integrin recycling. Furthermore, rhodamine-labeled C16 colocalized with activated β1 integrins. Flow cytometry analysis showed that C16 increased β1 integrin activation starting at 1 h of treatment. In summary, our results suggest that after interacting with the cell membrane and activating β1 integrins, breast cancer cells internalize peptide C16, which plays a role in β1 integrin turnover.

Mycotoxins are toxic secondary metabolites produced predominantly by fungal genera, such as Aspergillus, Fusarium, and Penicillium, and represent major foodborne contaminants responsible for chronic human exposure worldwide. While aflatoxin B1 (AFB1) is a well-established hepatocarcinogen, increasing evidence indicates that multiple mycotoxins contribute to tumorigenesis across diverse organ systems through shared biochemical and molecular mechanisms. At the molecular level, mycotoxins undergo cytochrome P450-mediated bioactivation, generating reactive intermediates that induce DNA adduct formation, oxidative stress, genomic instability, and disruption of redox homeostasis. These events converge on dysregulation of key signaling pathways governing cell-cycle control, apoptosis, immune surveillance, and epigenetic regulation, including aberrant DNA methylation, histone modification, and non-coding RNA expression. Importantly, emerging data support a "dual-hit" paradigm in which mycotoxin exposure synergizes with oncogenic viral infections, such as hepatitis B virus (HBV), human papillomavirus (HPV), and Epstein-Barr virus (EBV), amplifying genotoxic stress, immune evasion, and epigenetic instability. This review synthesizes current mechanistic insights into mycotoxin-induced carcinogenesis, emphasizing molecular toxicological endpoints that link exposure to cancer risk. In addition, advances in biosensing, detoxification, and preventive strategies are discussed, highlighting the need for mechanism-driven interventions to mitigate mycotoxin-associated carcinogenicity and its public health burden.

M2-like macrophages and CD8+T cells are key immune components that influence tumor behavior and treatment response. Ubiquitin-specific protease 32 (USP32) is established as a key oncogenic factor in gastric cancer (GC). This study aimed to investigate the role of USP32 in regulating M2 macrophage polarization and CD8+T cell dysfunction in GC. Macrophages derived from THP1 cells (THP1-M0) or CD8+T cells were co-cultured with transfected AGS and HGC-27 GC cells. The proportion of CD206+ M2 macrophages and the apoptosis of CD8+T cells were assessed by flow cytometry. Cell invasion was analyzed by transwell assay. The interaction between USP32 and death-associated protein kinase 1 (DAPK1) was verified by GST pull down and Co-immunoprecipitation (Co-IP) experiments. The effect on tumor growth was tested by subcutaneous xenograft studies. USP32 and DAPK1 were overexpressed in GC tissues and cell lines. Mechanistically, USP32 stabilized DAPK1 protein through deubiquitination. DAPK1 downregulation reversed USP32-mediated enhancement in GC cell invasion, macrophage M2 polarization, and CD8+T cell apoptosis in vitro. USP32 depletion exhibited an in vivo anti-growth effect on AGS subcutaneous xenografts. This study identifies the USP32/DAPK1 cascade as a crucial regulator of M2 macrophage polarization and CD8+T cell apoptosis in GC, providing a novel mechanistic link between post-translational regulation and tumor immune evasion.

Deltex E3 ubiquitin ligase 3L (DTX3L) is a well-established ubiquitin ligase implicated in various cancers, but its role in nasopharyngeal carcinoma (NPC) progression remains elusive. In this study, we confirmed for the first time that DTX3L was highly expressed in C666-1 and NPC/HK1 NPC cells. DTX3L overexpression promoted NPC cell proliferation, invasion, and migration. Conversely, DTX3L knockdown suppressed these malignant phenotypes. Notably, DTX3L activated the β-catenin pathway, as evidenced by increased β-catenin nuclear translocation, increased transcriptional activity, and elevated expression of its downstream target c-Myc. Mechanistically, we identified an upstream regulatory axis in which the oncogenic long noncoding RNA H19 acted as a molecular sponge for miR-423-5p, thereby alleviating the miR-423-5p-mediated repression of DTX3L. Dual-luciferase reporter assays confirmed that miR-423-5p directly targeted both DTX3L and H19, supporting the presence of a competitive endogenous RNA (ceRNA) network. Furthermore, rescue experiments demonstrated that DTX3L knockdown largely abolished the proliferative advantage conferred by H19 overexpression. Collectively, the results of our study revealed that the H19/miR-423-5p/DTX3L axis is a novel ceRNA-driven mechanism that promotes NPC progression via activation of β-catenin signaling.

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality globally; however, the molecular drivers remain unclear. Dysregulated cholesterol metabolism is a hallmark of HCC and contributes to tumor progression. The Niemann-Pick type C1 protein (NPC1), a lysosomal cholesterol transporter, is overexpressed in cancers; however, its oncogenic mechanisms in HCC remain unclear. In this study, we identified NPC1 as a critical regulator of HCC progression through dual mechanisms involving p53 destabilization and modulation of cholesterol metabolism. Analysis of the clinical data revealed that NPC1 was significantly upregulated in HCC tissues and correlated with poor prognosis. Functional studies have demonstrated that NPC1 silencing suppresses HCC cell proliferation, both in vitro and in vivo. Mechanistically, NPC1 interacts with deubiquitinase ubiquitin-specific protease 7 (USP7), disrupting its binding to p53 and enhancing p53 ubiquitination and proteasomal degradation. Concurrently, NPC1 modulates cholesterol synthesis and distribution via the p53-SREBP2 axis, and p53 knockdown reverses the cholesterol reduction caused by NPC1 silencing. The pharmacological activation of p53 reversed the decrease in cholesterol levels mediated by the overexpression of NPC1. These findings reveal that NPC1 is a multifaceted oncoprotein in HCC, linking cholesterol metabolism to p53 regulation and highlighting its potential as a therapeutic target for HCC intervention.

S-palmitoylation, a reversible lipid-based post-translational modification, is notably elevated in colorectal cancer (CRC) due to common lipid metabolism disorders. It has been reported to play crucial roles in regulating membrane composition, cell proliferation, and metastasis in various malignancies such as pancreatic and breast cancers. However, its role in the progression of CRC remains poorly understood. ZDHHC9, a member of the palmitoyl transferase family, is significantly upregulated in CRC patients and correlates with poor prognosis. Knockdown of ZDHHC9 impairs CRC cell proliferation and migration both in vitro and in vivo. RNA sequencing revealed that ZDHHC9 depletion markedly downregulates the cAMP signaling pathway. Mechanistically, ZDHHC9 knockdown impairs ADCY4 activity by reducing S-palmitoylation of KLF5 at cysteine 438, thereby modulating the ZDHHC9/KLF5/ADCY4 axis and downstream cAMP/PKA/CREB signaling to influence CRC cell proliferation and migration. Our findings demonstrate that ZDHHC9 promotes CRC progression by regulating intracellular cAMP levels through KLF5 palmitoylation, providing a novel therapeutic perspective targeting palmitoylation in CRC. The mechanism diagram of this study. ZDHHC9 mediates palmitoylation of KLF5 at cysteine 438, thereby enhancing ADCY4 activity and increasing intracellular cAMP levels. This elevation in cAMP promotes PKA and phosphorylation of CREB, ultimately activating the cAMP/PKA/CREB signaling pathway, which contributes to the regulation of CRC cell proliferation, migration, and resistance to 5-FU. (Created with BioRender.com).

Malignant cancers exhibit distinct lipid metabolic features that support tumor initiation and progression. Glioblastoma (GBM) is an aggressive brain tumor driven by GBM stem cells (GSCs), which are responsible for tumor development and therapy resistance. However, effective treatments targeting vulnerable metabolic pathways in GSCs have not yet been developed. Here, we demonstrate that the ATP-binding cassette transporter A3 (ABCA3) maintains lipid metabolic balance in GSCs. ABCA3 is highly expressed in GSCs, where lipid biosynthesis is particularly active. Knocking down ABCA3 significantly reduces cell growth, self-renewal, viability, and tumor growth after intracranial implantation. These changes are caused by a profound disruption of lipid metabolic balance, as demonstrated by RNA sequencing and liquid chromatography-time-of-flight mass spectrometry, which revealed widespread alterations in lipid metabolism genes and lipid composition. Mechanistically, ABCA3 knockdown inhibits sterol regulatory element-binding protein 1 (SREBP1) signaling by accumulating acylcarnitines (ACs) caused by phospholipid breakdown. The increased ACs induce the production of mitochondrial reactive oxygen species, which activate adenosine monophosphate-activated protein kinase (AMPK), resulting in the inhibition of SREBP1 signaling and reduced GSC fitness. Overall, these findings suggest that ABCA3 maintains lipid metabolic balance in GSCs, and disrupting this function triggers AMPK-dependent suppression of SREBP1 signaling.

Acute myeloid leukemia (AML) remains challenging to treat due to extensive genetic heterogeneity, high relapse rates, and treatment-related toxicity. Although drug combinations offer therapeutic promise, their selection is often empirical. Here, we introduce Combinatorial Proteome Integral Solubility/Stability Alteration analysis (CoPISA), a high-throughput proteomics workflow that captures protein solubility/stability alterations uniquely induced by drug combinations. We applied CoPISA to two rationally designed AML drug pairs, LY3009120-sapanisertib (LS) and ruxolitinib-ulixertinib (RU), previously identified as the most effective and least toxic combinations among many candidates and validated in AML cell lines, patient-derived samples and zebrafish xenograft models. We uncovered an emergent mechanism termed "conjunctional targeting", in which combinatorial drug action induces combination-exclusive protein targets consistent with an AND-gate logic model. LS-specific converged on SUMOylation, chromatin condensation, and VEGF-linked adhesion, while RU-specific targets disrupted DNA-damage checkpoints, mitochondrial bioenergetics, and RNA-splicing. Post-translational modification analysis revealed combination-induced acetylation, methylation, and phosphorylation of key AML proteins, including NPM1. Network analysis demonstrated that a substantial fraction of AML-associated proteins targeted by CoPISA are unique to combinations, including DNMT3A, NPM1, and TP53. By uncovering a mechanistic layer beyond classical synergy, CoPISA provides a robust framework for the precision-guided design of combinatorial therapies in heterogeneous cancers.

High-grade serous ovarian cancer (HGSOC) is characterized by a complex, immunosuppressive tumor microenvironment (TME) that contributes to poor clinical outcomes and resistance to therapy. To replicate the human TME in vitro, we develop a 3D pentaculture model incorporating five human cell types: malignant HGSOC cells and primary fibroblasts, mesothelial cells, adipocytes, monocytes. Monocytes differentiate into macrophages without exogenous cytokines in the pentacultures. Bulk RNA sequencing and deconvolution with single-cell RNA data from patient biopsies reveal that macrophage clusters within the pentacultures replicate those found in human HGSOC metastases, and proportions of individual clusters vary according to the malignant cell line. The pentacultures enable detailed analysis of malignant cell-macrophage interactions and highlight the influence of malignant cell genomic, transcriptomic and proteomic heterogeneity on the TME. Furthermore, targeting of "do-not-eat-me" signals with anti-CD47 and anti-CD24 monoclonal antibodies demonstrate differential effects on macrophage activity and cancer cell viability, again depending on the individual malignant cell line. Real-time microscopic monitoring of the pentacultures confirms dynamic modulation of macrophage behavior. We conclude that this pentaculture model offers a platform to study malignant cell control of TME elements and in particular their interactions with myeloid cells.