Search Results (3,302)

Introduction/Objectives: Implicated in the regionalization of the anterior central nervous system (CNS) and the progression of pallial astrogliogenesis, the transcription factor gene EMX2 has been reported to be specifically silenced in several tumors, both non-neural and neural. Based on that, its overexpression has been proposed as a tool for the treatment of a subset of these malignancies, including glioblastoma. Here we tested this proposal. Methods/Results: We found that Emx2 overexpression in human glioblastoma cells transplanted into the striatum of immunotolerant mice significantly prolonged animals’ survival, outperforming their treatment by temozolomide. However, this approach did not eradicate the tumor, because of in vivo silencing of the therapeutic Emx2 transgene. Notably, silencing of this transgene (or of a reporter designed to monitor its competence to be expressed) also occurred during long-term in vitro culture of GBM cells and was exacerbated by coculture with murine pallial glia under hypoxic conditions. Remarkably, partial insulation of the transgene and the use of a specific combination of epigenetic drugs substantially counteracted the progressive activatability decline undergone by the Emx2-transgene expression reporter, suggesting a promising strategy for overcoming a major limitation of Emx2-based therapy for GBM. Conclusions: An Emx2-encoding transgene, protected by insulation and appropriate epigenetic drugs, can provide a substantial benefit in experimental therapy for glioblastoma. Full article

Background/Objectives: Structured cytopathology reporting systems attach an evidence-based risk of malignancy (ROM) to every diagnostic category for the thyroid, salivary gland, urinary tract, lung and other sites. None exists for the oral cavity, and the ROM of an oral brush cytology category has not, to our knowledge, been reported. Methods: All oral liquid-based cytology specimens accessioned at one tertiary dental hospital between 2015 and 2024 were reviewed; those with a site-concordant histopathological diagnosis obtained within 90 days formed the analytic cohort (n = 137). Cytological reports were recovered from primary laboratory records and assigned to a five-tier ordinal scale. The target condition was malignancy or high-grade dysplasia, and diagnostic indices were calculated at every ordinal threshold with Wilson 95% confidence intervals. Results: ROM rose monotonically: 15.6% (95% CI 8.7–26.4) for negative, 53.3% (30.1–75.2) for atypical favoring reactive, 76.9% (61.7–87.4) for atypical cells of undetermined significance (ACUS), 87.5% (52.9–97.8) for atypical favoring neoplastic and 100% (74.1–100) for malignancy. The ACUS value far exceeded that of the similarly named tier in the thyroid (22%) and salivary gland (30.5%) systems and fell within the range spanned by their suspicious-for-malignancy tiers (74% and 83.8%). Malignancy was never reported in a histologically benign or low-grade lesions. Sensitivity at the ACUS threshold was 25.0% in keratotic against 79.3% in non-keratotic lesions (p = 0.004). Conclusions: These are, to our knowledge, the first category-specific ROM estimates for oral brush cytology, and they identify a nomenclature problem: a category named for the pathologist’s uncertainty carried a risk equivalent to suspicious for malignancy elsewhere; while a negative report was followed by disease in one lesion in six. Any oral reporting system should attach an explicit ROM to each category; our exploratory data further suggest that the surface keratinization of the sampled lesion should be recorded. Full article

Medulloblastoma is increasingly best understood not simply as a cell-autonomous embryonal tumour, but as a family of molecularly and anatomically distinct diseases shaped by the developing cerebellum and posterior fossa. The four principal molecular subgroups—WNT, sonic hedgehog, Group 3 and Group 4—differ not only in genetics and outcome, but also in barrier permeability, immune composition, stromal crosstalk and routes of dissemination. Histopathological variation likewise reflects differences in tumour architecture, neuronal maturation and cytological aggressiveness that intersect with, but are not reducible to, molecular subgroup. This review takes a microenvironment-centred view of medulloblastoma biology. It asks how non-malignant cells, fluid compartments and physical interfaces—including immune cells, the choroid plexus and cerebrospinal fluid, the blood–brain and blood–tumour barriers, astrocytes and experimental model systems—modify tumour initiation, progression, therapy response and relapse. The central argument is that the medulloblastoma tumour microenvironment is not merely sparse or passive; instead, pathological, spatial and molecular analyses of human tumours, complemented by models that preserve the relevant niche, reveal a developmentally patterned, subgroup-specific and therapeutically tractable ecosystem. Full article

Titanium dioxide (TiO₂) is a widely studied semiconductor whose interfacial redox properties strongly influence its photocatalytic and biological performance. In this work, TiO₂ nanomaterials were surface-functionalized with green tea waste extract (GT) and subsequently impregnated with silver to obtain redox-active nanocomposites with tunable optical and biological properties. HPLC, FTIR, diffuse reflectance spectroscopy (DRS), and density functional theory (DFT) analyses demonstrated the formation of an organic–inorganic interface through adsorption of green tea-derived ligands. DFT calculations revealed complementary interfacial roles of the adsorbed constituents, with epigallocatechin gallate (EGCG) inducing interfacial charge-transfer (ICT) states that enable visible-light absorption, as reflected by the decrease in the apparent optical bandgap from ~3.48 eV for pristine TiO₂ to ~1.83 eV for TiO₂/EGCG. ICP-OES analysis further quantified the Ag loading in TiO₂/GT/Ag at 5.1 wt%. Electron paramagnetic resonance (EPR) experiments demonstrated that GT functionalization shifts the interfacial redox balance toward radical scavenging by suppressing excessive radical generation, whereas silver incorporation partially restores oxidative pathways, particularly under irradiation. These differences in interfacial redox behavior directly translate into distinct biological responses. TiO₂/GT/Ag showed the strongest antimicrobial activity, with visible-light enhancement observed predominantly against Staphylococcus aureus, while TiO₂/GT/Ag reduced H₂O₂-induced oxidative stress in non-malignant cells and promoted intracellular reactive oxygen species generation in cancer cells. These findings demonstrate that engineering the organic–inorganic interface through plant-derived ligands and silver incorporation provides an effective strategy for tuning the interfacial redox properties and light-responsive biological performance of TiO₂-based nanomaterials, thereby expanding their potential for antimicrobial and biomedical applications. Full article

5-Methylcytosine (m⁵C) is a widespread mRNA modification that regulates gene expression and is frequently dysregulated in cancer. Here, we used phage display to identify peptides that bind an NSUN2 consensus RNA sequence in either its unmodified (UN2-RNA) or m⁵C-modified (MN2-RNA) form. A single peptide, un2p1 (TDYSTHRLSHSL), was selectively enriched against both targets. Biophysical analyses demonstrated that un2p1 binds this RNA sequence with high affinity and that m⁵C incorporation reduces binding by approximately ninefold, indicating that cytidine methylation acts as a biochemical switch for peptide–RNA recognition. Sequence alignments mapped un2p1 to a conserved structural loop and catalytic domain region within NSUN2, suggesting that the peptide functions as a molecular mimic of the endogenous RNA-recognition interface. In A549 lung adenocarcinoma cells, un2p1 treatment reduced global m⁵C levels and downregulated the oncogenic chaperonin CCT5, consistent with disruption of NSUN2-dependent Wnt/β-catenin signaling. These effects were attenuated in non-malignant HEK293 cells, which exhibited a compensatory increase in m⁵C levels and preserved viability. Together, these findings identify un2p1 as a sequence-specific, methylation-sensitive RNA-binding peptide that modulates the m⁵C epitranscriptome and selectively impairs lung cancer cell viability, highlighting a substrate-centric strategy for targeting NSUN2-mediated oncogenic pathways. Full article

Agmatine is a naturally occurring polyamine with context-dependent effects on cellular redox regulation, survival, and apoptosis; however, its effects in endometrial cancer cells remain poorly characterized. This exploratory in vitro study investigated cellular responses associated with agmatine sulfate exposure in Ishikawa endometrial adenocarcinoma cells, using human dermal fibroblasts (HDFs) as a non-cancerous, non-tissue-matched comparator. Metabolic activity was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay following exposure to 0.5–15 mM agmatine sulfate for 24 and 48 h. Based on the 48-h concentration-response analysis in Ishikawa cells, 4.1 and 6.5 mM were selected for subsequent analyses of intracellular reactive oxygen species (ROS), malondialdehyde (MDA), cleaved caspase-3, and p-AKT/p-JNK immunoreactivity. Agmatine sulfate produced concentration- and time-dependent reductions in MTT-derived metabolic activity in both cell lines. In Ishikawa cells, 4.1 and 6.5 mM agmatine sulfate significantly increased ROS, MDA, and cleaved caspase-3 levels, accompanied by decreased p-AKT and increased p-JNK immunoreactivity. In HDF cells, ROS levels increased at both concentrations, whereas significant increases in MDA, cleaved caspase-3, and p-JNK were observed only at 6.5 mM; p-AKT immunoreactivity remained unchanged. These findings indicate that agmatine sulfate exposure is associated with reduced metabolic activity, increased oxidative stress and apoptosis-associated responses, and changes in AKT/JNK-related signalling in Ishikawa cells. However, the responses observed in HDF cells do not support a conclusion of tumour-selective activity. Further studies using additional endometrial cancer cell lines, tissue-matched non-malignant controls, and targeted experimental approaches are warranted to clarify the biological relationships among these responses. Full article

Background: Disseminated invasive aspergillosis (IA) is an uncommon but devastating manifestation of disease, particularly when the central nervous system (CNS) is involved. The clinical features and outcomes of CNS dissemination, compared with isolated invasive pulmonary aspergillosis (IPA) and other disseminated forms, remain poorly characterized. Methods: Using an institutional database of 1157 cancer patients diagnosed with IA between 1993 and 2024, we identified 43 patients with disseminated IA, defined as infection involving ≥2 non-contiguous organ systems, including 8 with CNS involvement. Patients with disseminated IA were matched 1:3 to patients with IPA based on year of diagnosis. We compared clinical features, antifungal treatment, and outcomes between: (1) CNS-disseminated IA and IPA, and (2) CNS-disseminated IA and other forms of disseminated IA. Results: Baseline characteristics, including hematologic malignancy subtype, hematopoietic stem cell transplantation status, neutropenia, and antifungal prophylaxis, were similar across groups. Aspergillus fumigatus was the predominant species in all cohorts. No patients with CNS-disseminated IA achieved clinical response at end of therapy, versus 35% with IPA (p = 0.05) and 30% with other disseminated IA (p = 0.17). Twelve-week IA-associated mortality was significantly higher in CNS-disseminated IA than in IPA (88% vs. 45%, p = 0.028) and was higher than in other disseminated IA (88% vs. 46%, p = 0.05). Combination antifungal therapy was more frequently used in CNS-disseminated IA than in IPA (63% vs. 31%) and other disseminated IA (63% vs. 43%), while rates of ICU admission and mechanical ventilation were similar across groups. Conclusions: CNS involvement in disseminated IA defines a distinct, high-risk phenotype, with profoundly reduced treatment response and survival despite comparable baseline characteristics. Given the small number of CNS cases and the predominance of cases diagnosed during earlier study periods, these findings should be interpreted with caution. Although combination antifungal therapy was used more frequently in CNS-disseminated IA, no significant outcome benefit was observed, underscoring the need for improved therapeutic approaches for CNS aspergillosis in immunocompromised hosts. Full article

Background: Axillary lymph node (ALN) fine-needle aspirations (FNAs) can guide the next steps in the management of patients with invasive breast carcinoma (BC). The objective is to evaluate the diagnostic performance of ALN FNA in the detection of metastatic breast carcinoma. Methods: We evaluated ALN FNAs from 2020 to 2024 to determine the risk of malignancy (ROM) of different cytologic diagnostic categories using a general WHO cytology reporting system framework (insufficient, benign, atypical, suspicious, and malignant). Results: A total of 290 axillary lymph node FNAs from patients with newly diagnosed invasive breast carcinoma were identified, of which 226 (78%) had histologic and/or adequate clinical follow-up. Cytologic diagnoses included six (2.1%) non-diagnostic, 99 (34.1%) benign, two (0.7%) atypical, seven (2.4%) suspicious for malignancy, and 176 (60.7%) malignant cases. The risk of malignancy (ROM) was 50.0% for non-diagnostic, 17.0% for benign, 100% for atypical, 85.7% for suspicious, and 99.2% for malignant diagnoses. Most false-negative benign cases were attributed to sampling limitations rather than interpretive error, with five cases showing only a single sentinel lymph node metastasis measuring <5 mm. Indeterminate diagnoses were most commonly associated with scant cellularity, acellular cell blocks, obscuring blood or fibrin, and equivocal immunohistochemistry. Excluding cases without histologic follow-up, ALN FNA demonstrated a sensitivity of 86.5%, specificity of 97.4%, positive predictive value of 98.5%, negative predictive value of 79.2%, and overall accuracy of 90.3% for the detection of metastatic breast carcinoma (OR = 243.2, Fisher’s exact test, p < 0.0001). Conclusion: ALN FNA is a highly reliable method for confirming axillary lymph node metastasis in invasive breast carcinoma when malignant cells are identified, with a ROM of 99% for the malignant category. The suspicious category was also strongly associated with malignancy (ROM 86%). However, the 17% ROM observed in the benign category indicates that a negative FNA does not reliably exclude metastatic disease, and surgical nodal staging may still be warranted in clinically or radiologically suspicious cases. These findings support the clinical applicability of the WHO System for Reporting Lymph Node Cytopathology and provide breast carcinoma-specific ROM estimates for this clinical setting. Full article

Background: Pleural effusion is a common clinical presentation in both benign and malignant conditions. The advent of liquid biopsy allowed new possibilities for non-invasive molecular profiling using body fluids. Pleural fluid, by virtue of its proximity to thoracic malignancies and its rich tumour-derived content, represents a particularly compelling matrix for liquid biopsy analysis. This review synthesises current evidence regarding the diagnostic, predictive, and prognostic utility of pleural liquid biopsy in clinical medicine. Methods: A narrative review was conducted using PubMed, MEDLINE, and EMBASE databases. Search terms included combinations of “pleural effusion”, “liquid biopsy”, “circulating tumour DNA”, “circulating tumour cells”, “exosomes”, “next-generation sequencing”, and “biomarkers”. Priority was given to original research articles, systematic reviews, and meta-analyses published between 2013 and 2026. Results: Pleural fluid contains a diverse repertoire of tumour-derived analytes, including cell-free and circulating tumour DNA (ctDNA), circulating tumour cells (CTCs), exosomes, and soluble proteins. These biomarkers enable molecular characterisation of underlying malignancies with sensitivity that often exceeds that of plasma-based liquid biopsy and complements histological tissue biopsy. Detection of actionable mutations, including EGFR, ALK, KRAS, and BRAF alterations, directly informs targeted therapy selection. Furthermore, serial sampling facilitates real-time monitoring of therapeutic resistance, disease progression, and clonal evolution. Conclusions: Pleural liquid biopsy offers a minimally invasive, reproducible, and clinically informative approach to molecular profiling in patients with pleural disease, particularly those with thoracic malignancies. Despite existing challenges in standardisation and analytical sensitivity, its integration into routine clinical pathways holds significant promise for advancing personalised oncological care. Multi-omics integration and artificial intelligence may further consolidate its role in therapeutic decision-making. Full article

Non-small-cell lung cancer (NSCLC) treatment is hampered by its complex pathogenesis and high heterogeneity. N6-methyladenosine (m⁶A) represents the most common post-transcriptional modification regulating RNA stability and function in eukaryotes. This methylation is catalyzed by methyltransferase complexes, with METTL14 being the core catalytic subunit. Abnormal expression of lncRNA MSTRG.292666.16 is related to poor prognosis of NSCLC. However, the mechanism by which it regulates NSCLC progression through m⁶A modification remains unclear. We employed cell function experiments, molecular mechanism analysis, RNA interaction experiments, and a nude mouse tumor model to explore the roles of METTL14-mediated MSTRG.292666.16 m⁶A modification in NSCLC and the potential MAPK signaling pathway involved. METTL14 was significantly upregulated in NSCLC cell lines and promoted m⁶A modification of MSTRG.292666.16 by forming a stable association with it. METTL14 knockdown significantly inhibited the viability, migration and invasion of A549 cells and promoted apoptosis, whereas MSTRG.292666.16 overexpression reversed these effects. Mechanistically, METTL14 upregulated the expression of MSTRG.292666.16 through m⁶A modification, thereby activating the MAPK pathway (manifested as elevated levels of MAPK8IP3 and p-ERK1/2). The use of a selective p38 MAPK inhibitor SB203580 stimulated the tumor-suppressive effect of METTL14 knockdown, whereas the activator U-46619 reversed it. In vivo experiments confirmed that METTL14 knockdown significantly inhibited tumor growth, whereas MSTRG.292666.16 overexpression partially restored the malignant phenotype of the tumor, which was associated with MAPK pathway activation. This study revealed that METTL14-dependent m⁶A modification of MSTRG.292666.16 may act as an upstream driver to activate the MAPK cascade and facilitate NSCLC progression. These findings clarify a key epitranscriptomic regulatory mechanism driving NSCLC development and offer preliminary molecular clues for exploring potential therapeutic targets in subsequent clinical NSCLC research. Full article

Background: Oral leukoplakia is a clinically important oral potentially malignant disorder with variable risk of progression to oral squamous cell carcinoma. Histopathological grading of oral epithelial dysplasia remains the primary method for risk assessment; however, its predictive accuracy is limited by sampling variability, lesion heterogeneity, and interobserver variation. DNA aneuploidy and abnormal DNA ploidy status have therefore been investigated as objective biomarkers of genomic instability and potential predictors of malignant transformation. Aim: This narrative review evaluates whether DNA aneuploidy or abnormal DNA ploidy status predicts malignant transformation of oral leukoplakia to oral squamous cell carcinoma compared with diploid DNA status, and whether DNA ploidy should be interpreted as a stand-alone prognostic marker or as part of combined risk-assessment models. Materials and Methods: A structured PubMed/MEDLINE search (last performed on 12 July 2026) identified 162 records using terms related to oral leukoplakia, oral potentially malignant disorders, DNA ploidy, DNA aneuploidy, cytometry, malignant transformation, and oral squamous cell carcinoma. After screening and full-text eligibility assessment, 45 studies were included and organised according to their evidential role as core prognostic studies, PVL-specific subgroup evidence, and supporting/background evidence addressing diagnostic relevance, dysplasia correlation, treatment monitoring, genomic instability, and methodological development. Results: Most core prognostic studies showed that DNA aneuploidy, abnormal DNA content, or chromosomal instability was associated with increased malignant transformation risk compared with diploid or non-aneuploid status. Several studies reported higher transformation rates, hazard ratios, or improved prediction when DNA ploidy was combined with dysplasia grading, lesion site, clinical heterogeneity, or other biomarkers. However, predictive performance varied across studies, and DNA ploidy alone often showed modest sensitivity, specificity, or positive predictive value. Diploid or non-aneuploid status appeared more useful for identifying lower-risk lesions, although it did not completely exclude malignant transformation. PVL evidence suggested frequent aneuploidy and aggressive genomic behaviour, but DNA ploidy appeared less reliable for discriminating risk within PVL. Conclusions: DNA aneuploidy and abnormal DNA ploidy status are meaningful markers of genomic instability and are associated with increased malignant transformation risk in oral leukoplakia. However, when used alone, DNA ploidy often shows only modest predictive performance, and prediction improves consistently when it is combined with dysplasia grading and other clinicopathological factors. Their strongest clinical value is therefore as adjunctive biomarkers within combined clinicopathological and molecular risk models, rather than as stand-alone replacements for histopathological assessment; routine clinical implementation is not yet supported by the current evidence. Full article

Clear cell renal cell carcinoma (ccRCC) is a highly immune-evasive malignancy that exhibits limited and often transient responses to immune checkpoint inhibitors (ICIs) and remains a major challenge for emerging cellular immunotherapies, including chimeric antigen receptor (CAR)-T cells. A defining feature of ccRCC, largely driven by von Hippel–Lindau (VHL) deficiency, is persistent activation of the transforming growth factor-β1 (TGF-β1) and hypoxia-inducible factor (HIF) signaling network. Acting as a central immunometabolic regulatory axis, TGF-β1/HIF promotes angiogenesis, metabolic reprogramming, epigenetic dysregulation, and immune escape through coordinated induction of immunosuppressive mediators, including PD-L1, VEGF, and CTLA-4, resulting in impaired T-cell infiltration, functional exhaustion, and resistance to immunotherapy. Preclinical studies have demonstrated that pharmacologic-dose Seleno-L-methionine (SLM) and its active metabolite, methylseleninic acid (MSA), suppress TGF-β1 and both HIF-1α and HIF-2α, leading to downregulation of multiple downstream immunosuppressive pathways at pharmacologically achievable, non-toxic concentrations. In addition to enhancing the antitumor activity of chemotherapy and VEGF-targeted agents, accumulating evidence suggests that SLM exerts broad immunologic, metabolic, and epigenetic effects that may overcome key mechanisms of therapeutic resistance. This review synthesizes current mechanistic and translational evidence supporting the repurposing of SLM as a first-in-class pleiotropic immunomodulatory agent. Using ccRCC as a model of TGF-β1/HIF-driven immune resistance, we discuss how simultaneous targeting of this central regulatory axis may restore immune surveillance, improve T-cell fitness and trafficking, enhance responses to ICIs and CAR-T cell therapy, and provide a mechanistically rational strategy for the treatment of advanced solid tumors. Full article

Background/Objectives: Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited targeted therapeutic options. Although the anticancer and pro-apoptotic properties of boswellic acids have previously been reported, the interaction profile of chemically defined β-boswellic acid (BA) with doxorubicin (DOX) remains insufficiently characterised in the murine 4T1 TNBC-like model. This study quantitatively evaluated BA–DOX drug interactions using different reference models and characterised the cytotoxic and apoptosis-associated cellular phenotype accompanying combined exposure. Methods: Cytotoxicity was assessed using the MTT assay and BA–DOX interactions were evaluated using the Chou–Talalay combination index (CI), highest single-agent (HSA) and Bliss independence models. Apoptosis and cell-cycle distribution were analysed by flow cytometry and mitochondrial membrane potential was assessed by JC-1 staining. Caspase-3/7 activity, RT-qPCR, live/dead Calcein-AM/PI staining, 4′,6-Diamidino-2-phenylindole (DAPI) nuclear staining, and cytokine measurements were also performed. Gene Ontology (GO), Kyoto Encyclopaedia of Genes and Genomes (KEGG), and STRING-based protein–protein interaction (PPI) analyses were used to explore putative molecular pathways associated with experimental findings. Results: The 48 h selectivity index of BA was 1.28, indicating only modest differential cytotoxicity between 4T1 cells and HaCaT keratinocytes under the experimental conditions rather than definitive cancer-cell selectivity. Drug-interaction analyses revealed concentration- and model-dependent effects, with the Chou–Talalay analysis indicating synergism in selected intermediate and higher concentration pairs. Under the selected phenotypic-characterisation condition, BA + DOX produced a greater apoptotic response than either single treatment, accompanied by increased G2/M and Sub-G1 fractions, mitochondrial membrane depolarisation, and increased caspase-3/7 activity. This treatment condition was not included in the drug-interaction analysis and was therefore not interpreted as a pharmacologically validated synergistic combination. RT-qPCR demonstrated increased mRNA expression of Bax, Casp3, and Casp9, decreased mRNA expression of Bcl2, and a marked increase in the Bax/Bcl2 mRNA ratio. Calcein-AM/PI and DAPI analyses further demonstrated increased cell death and apoptotic nuclear alterations. The measured concentrations of TNF-α and IL-6 in culture supernatants were lower after BA + DOX treatment, whereas IL-10 remained unchanged; however, these cytokine measurements were not normalised to viable cell number and therefore require cautious interpretation. Exploratory bioinformatic analyses identified predicted associations with apoptosis-, mitochondrial-, and cell-cycle-related processes and pathways; however, these database-derived findings were considered hypothesis-generating and not evidence of BA-dependent target engagement or pathway activation. Conclusions: Combined BA and DOX exposure produced greater cytotoxic and apoptosis-associated responses than either single treatment in 4T1 cells, whereas formal drug-interaction classifications varied according to concentration and analytical model. The accompanying changes in mitochondrial membrane potential, caspase-3/7 activity, and apoptosis-related gene expression describe a treatment-associated cellular phenotype but do not identify a direct molecular target of BA or establish a causal molecular mechanism. The findings also do not demonstrate TNBC-specific selectivity. Further studies using additional breast cancer and tissue-matched non-malignant models, together with direct target-engagement and functional pathway-validation approaches, are required. Full article

Background/Objectives: Peritoneal dissemination of malignancies leads to poor prognoses, and no effective treatment currently exists. The difficulty of treating such malignancies is likely because systemically administered drugs cannot easily target malignant cells in the abdominal cavity. High intraperitoneal drug retention, non-toxicity towards normal tissues, and successful targeting of malignant cells are important for an effective therapy. The aim of this study was to evaluate an intraperitoneally administered astatine-labeled, integrin-targeted nanodrug, mPEG(Mn:350)-S-AuNP[²¹¹At]-c[RGDfK(C)] ([²¹¹At]AuNP@PEG/RGD), with respect to its kinetics, therapeutic efficacy, and safety. Methods: C6 rat glioma cells (10⁷), and BxPC3 (10⁷) and PANC-1 (10⁷) human pancreatic cancer cells were seeded intraperitoneally into nude mice, and [²¹¹At]AuNP@PEG/RGD (0.979 ± 0.194 MBq for C6 models (n = 3), 1.139 ± 0.035 MBq for BxPC3 models (n = 10), and 1.308 ± 0.039 MBq for PANC-1 models (n = 10) per mouse) or saline were intraperitoneally administered 4–7 days later. Cytotoxicity against malignant cells, pharmacokinetics after administration, therapeutic efficacy, and safety in abdominal organs were evaluated. Results: Intraperitoneally administered [²¹¹At]AuNP@PEG/RGD accumulated exclusively in the peritoneal cavity for a long period of time and showed minimal systemic diffusion through the blood. In the C6 model, the intraperitoneal tumor mass was significantly lower in the treated group compared with that of the controls (p = 0.05). For the BxPC3 (median survival time: control/treated = 41/65 days, p < 0.001) and PANC-1 (median survival time: control/treated = 19/35 days, p < 0.001) peritoneal dissemination models, survival analysis revealed that [²¹¹At]AuNP@PEG/RGD significantly prolonged overall survival. Although transient weight loss, leukopenia, and thrombocytopenia were observed at one week post-administration, a short recovery trend was evident thereafter. One month after administration, no abnormalities were found in hematological tests or histological analyses of intra-abdominal organs. Conclusions: The intraperitoneal administration of astatine-labeled integrin-targeted [²¹¹At]AuNP@PEG/RGD nanoparticles showed promising findings in terms of safety and efficacy for treating peritoneally disseminated malignant tumors. Full article

Background/Objectives: Rosmarinic acid (RA) is a naturally occurring polyphenolic compound with promising anticancer activity; however, its potential to enhance the therapeutic efficacy of doxorubicin (DOX) against triple-negative breast cancer (TNBC) has not been comprehensively characterised. This study investigated the pharmacological interaction and associated cellular responses of RA combined with DOX in MDA-MB-231 breast cancer cells, while including HaCaT human keratinocytes as a non-malignant, non-mammary reference model for comparative cytotoxicity assessment. Methods: Cell viability was assessed using the MTT assay, and pharmacological interactions were evaluated using Chou–Talalay combination index (CI) and dose reduction index (DRI) analyses. Intracellular oxidative activity was evaluated using DCFH-DA fluorescence analyses together with N-acetyl-L-cysteine (NAC) modulation experiments. Apoptosis, cell-cycle distribution, mitochondrial membrane potential (JC-1), Caspase-9 immunocytochemistry, nuclear morphology (NucBlue staining), apoptosis- and proliferation-related gene expression (RT-qPCR; BAX, BCL2, CASP3, CASP9, TP53, CDKN1A, MKI67, and PCNA), and bioinformatic pathway analyses were performed to characterise cellular and molecular responses associated with the combined treatment. Results: After 48 h, RA exhibited IC₅₀ values of 188.4 ± 4.2 µM in MDA-MB-231 cells and 146.6 ± 6.8 µM in HaCaT cells, while DOX showed IC₅₀ values of 1.2 ± 0.08 µM and 2.6 ± 0.15 µM, respectively. The Chou–Talalay analysis demonstrated synergistic interactions in MDA-MB-231 cells, with CI values of 0.86, 0.72, and 0.64 at the effect levels of IC₂₅, IC₅₀, and IC₇₅, respectively; the model-derived DOX DRI value at Fa = 0.50 was 1.8. Combination treatment markedly increased intracellular DCFH-DA fluorescence, whereas NAC pretreatment attenuated this signal and partially restored cell viability, supporting a contributory role of intracellular oxidative stress in treatment-associated cytotoxicity. Increased intracellular oxidative activity coincided with mitochondrial membrane depolarisation, increased apoptotic cell death, accumulation of cells in the sub-G₁ phase, enhanced Caspase-9 immunoreactivity, and pronounced apoptotic nuclear alterations. RT-qPCR analysis demonstrated significant upregulation of BAX, CASP3, CASP9, and TP53, together with downregulation of BCL2, MKI67, and PCNA, resulting in a marked reduction in the BCL2/BAX mRNA expression ratio. A modest but non-significant increase in CDKN1A expression was also observed. Bioinformatic analyses further identified predicted associations with mitochondrial apoptosis and p53-associated signalling pathways. Conclusions: The RA + DOX combination showed synergistic cytotoxic activity in MDA-MB-231 cells, accompanied by increased intracellular oxidative activity, mitochondrial membrane depolarisation, apoptosis, cell-cycle perturbation, and changes in proliferation-related gene expression. These findings indicate associated cellular responses but do not establish a ROS-dependent mitochondrial apoptotic mechanism. The findings provide an exploratory in vitro basis for further investigation of the RA + DOX combination in additional TNBC models, non-malignant mammary epithelial cells, and appropriate in vivo systems. Full article