Search Results (1,752)

Triple-negative breast cancer (TNBC) is an aggressive and clinically heterogeneous breast cancer subtype characterized by the absence of estrogen receptor, progesterone receptor, and HER2 overexpression, limited targeted treatment options, early relapse, and frequent development of therapy resistance. Although TNBC often shows initial sensitivity to chemotherapy, durable responses are commonly undermined by the emergence of adaptive resistant cell states rather than solely by fixed genetic mutations. This review synthesizes the role of epigenetic plasticity as a central mechanism that enables TNBC cells to dynamically reprogram transcriptional identity, survive therapeutic stress, and transition between epithelial, mesenchymal, stem-like, immune-evasive, and drug-tolerant persister phenotypes. Key epigenetic mechanisms include aberrant DNA methylation, histone acetylation and methylation, BET/BRD4-dependent transcriptional regulation, EZH2-mediated repression, SWI/SNF-dependent chromatin remodeling, non-coding RNA networks, and three-dimensional genome reorganization. These processes regulate tumor suppressor silencing, DNA-damage repair, epithelial–mesenchymal plasticity, cancer stem-cell maintenance, metabolic adaptation, immune-checkpoint regulation, and minimal residual disease. The review also highlights the translational relevance of epigenetic biomarkers, including DNA methylation signatures, circulating epigenetic markers, chromatin-accessibility profiles, and single-cell epigenomic approaches for diagnosis, prognosis, therapy prediction, and monitoring resistance evolution. Finally, therapeutic strategies targeting epigenetic plasticity are discussed, including DNMT, HDAC, BET, EZH2, KDM, and LSD1 inhibitors, with emphasis on rational combination approaches involving chemotherapy, PARP inhibitors, immunotherapy, and metabolic targeting. Overall, epigenetic plasticity represents both a major driver of TNBC resistance and a therapeutically exploitable vulnerability, provided those future strategies account for tumor heterogeneity, adaptive cell-state transitions, biomarker-guided patient selection, and combination-based treatment design. Full article

Background: Triple-negative breast cancer (TNBC) remains a major cause of cancer mortality due to its aggressive behavior, metabolic adaptability, and high therapeutic resistance. Extracellular vesicles (EVs) within the tumor microenvironment contribute to tumor progression and metastasis by transferring pro-tumorigenic cargo. While conventional Tumor Treating Fields use high-frequency alternating fields to disrupt mitosis, low-energy triple-frequency bioelectromagnetic approaches remain poorly characterized. Methods: We evaluated a device–drug strategy combining triple-frequency low-intensity electromagnetic stimulation (EMS2: 396 Hz, 285 Hz, 528 Hz) with the pleiotropic drug Fingolimod (FTY720). Treatments were tested in MDA-MB-231 and ARM-G breast cancer cells, with Paclitaxel as a positive control. Cell proliferation was assessed by MTS assay, and extracellular vesicles were isolated following individual and combination treatments. Quantitative LC-MS/MS proteomics was used to characterize treatment-induced changes in EVs cargo. Results: EMS2 reduced proliferation in both cell lines and produced morphological changes consistent with altered cell-cycle progression. EMS2 alone triggered adaptive metabolic responses, whereas combination with Fingolimod suppressed these compensatory signatures. EVs proteomics revealed combination-specific alterations associated with mitochondrial stress, ER stress, NF-κB suppression, and autophagy-associated pathways. The combination also reduced levels of metastasis- and stroma-associated proteins, including Mitogen-Activated Protein Kinase 12 (MAPK12) and collagen-associated ECM components (Collagen Type I Alpha 1 Chain (COL1A1), Collagen Type VI Alpha 1 Chain (COL6A1), Collagen Type VI Alpha 3 Chain (COL6A3), and Matrilin 3 (MATN3)) in EVs. Bliss independence analysis identified a subset of metastasis-associated proteins suppressed in EVs beyond the level predicted by an additive model, an exploratory finding that will require further validation with dose–response and functional assays. Conclusions: Combined triple-frequency EMS2 and Fingolimod treatment altered the extracellular vesicle proteome, inducing signatures consistent with mitochondrial and endoplasmic reticulum stress, metabolic disruption, and reduced levels of metastasis-associated and stromal/ECM remodeling proteins, along with reduced proliferation. These findings suggest a coordinated anti-cancer effect of this tunable device–drug strategy, warranting further functional and in vivo validation to confirm therapeutic potential. Full article

Background/Objectives: Neoadjuvant therapy (NAT) may remodel the breast-cancer microenvironment. We evaluated paired changes in programmed death-ligand 1 (PD-L1), vascular endothelial growth factor (VEGF), and tumor-infiltrating lymphocytes (TILs) before and after NAT, and associations with the residual cancer burden (RCB) and survival. Methods: This retrospective study included 102 patients with residual invasive breast cancer after NAT: 34 with luminal B-like/HER2-negative, 34 with luminal B-like/HER2-positive, and 34 with triple-negative breast cancer (TNBC). PD-L1 was assessed using the 22C3 combined positive score, VEGF by the cytoplasmic staining intensity, and stromal TILs according to international recommendations. Results: The median tumor size decreased from 2.5 to 1.7 cm (p < 0.001), the PD-L1 CPS (combined positive score) from 6 to 5 (p = 0.039), and the TILs from 15% to 10% (p < 0.001), whereas VEGF shifted toward stronger staining (p = 0.03). In TNBC, the PD-L1 CPS decreased from 10 to 5 (p = 0.003) and the TILs from 20% to 8% (p < 0.001). Biomarkers were not associated with the RCB in the overall cohort. The initial tumor size predicted RCB II/III (OR: 2.60, p = 0.038). The overall survival differed by subtype (p < 0.001). Conclusions: NAT has induced subtype-dependent immune and angiogenic changes, supporting biomarker reassessment in residual disease. Full article

Objectives: Platinum-based neoadjuvant therapy can improve pathological response in triple-negative breast cancer (TNBC), but severe hematotoxicity may compromise treatment delivery. This study developed and validated a multitask learning framework to jointly predict pathological complete response (pCR) and severe hematotoxicity and to support benefit–hematotoxicity stratification. Methods: This multicenter retrospective study included 2060 consecutive patients with TNBC receiving platinum-based neoadjuvant therapy at three institutions. Patients were assigned to a training cohort (n = 1406), an internal validation cohort (n = 351), or an external validation cohort (n = 303). A multitask TabNet (MT-TabNet) model was developed using pretreatment clinical, pathological, imaging, laboratory, electrocardiographic, and planned treatment exposure variables to jointly estimate pCR and severe hematotoxicity. Model performance was evaluated using discrimination, calibration, decision curve analysis, and interpretability analyses. The predicted probabilities were further integrated into a utility-based framework for benefit–hematotoxicity stratification. Results: Overall, 639 patients (31.0%) achieved pCR, and 651 (31.6%) developed severe hematotoxicity. MT-TabNet achieved AUCs of 0.874, 0.842, and 0.819 for pCR prediction and 0.859, 0.846, and 0.818 for severe hematotoxicity prediction in the training, internal validation, and external validation cohorts, respectively. The model showed generally acceptable calibration and clinical net benefit. pCR prediction was predominantly associated with tumor-related characteristics, whereas severe hematotoxicity prediction was more strongly associated with planned treatment exposure and host-related laboratory indicators. Utility-based stratification showed progressively higher pCR rates and lower severe hematotoxicity rates from the low- to high-benefit groups across all three cohorts. Survival differed significantly among benefit groups for both progression-free survival (PFS) and overall survival (OS) in the training and internal validation cohorts; in the external validation cohort, OS differed significantly, whereas PFS showed a similar but nonsignificant trend. Conclusions: MT-TabNet enabled joint estimation of pCR and severe hematotoxicity in patients with TNBC receiving platinum-based neoadjuvant therapy. The utility-based framework integrated efficacy and toxicity predictions into clinically interpretable benefit–hematotoxicity stratification and warrants further prospective evaluation for individualized risk assessment and toxicity monitoring. 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: 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

Background: Solid tumors including glioblastoma (GBM), lung cancer (LC), and triple-negative breast cancer (TNBC) exhibit marked radioresistance driven by dysregulated cell-cycle control and genomic instability. Although CDK4/6 inhibitors suppress tumor proliferation, their radiosensitizing capacity is limited by persistent DNA repair. BUB1, a mitotic checkpoint kinase overexpressed in aggressive cancers, has emerged as a regulator of DNA damage signaling. We tested whether co-targeting BUB1 and CDK4/6 enhances radiosensitivity across solid tumors. Methods: GBM, LC, and TNBC cell lines were treated with BUB1 inhibitor BAY1816032, CDK4/6 inhibitors ribociclib and abemaciclib, and radiation. Proliferation, clonogenic survival, immunoblotting, and combination index analyses assessed cytotoxicity and synergy. CDK4/6-resistant models were generated to examine resistance. In vivo efficacy was evaluated using SUM159 xenografts. DNA damage and homologous recombination repair were measured by gH2AX, RAD51, RPA, BrdU foci and comet assay. The resection branchpoint was assessed by phospho-RPA, with ATR inhibition and BLM or EXO1 depletion testing resection dependence. Results: BUB1 inhibition increased cytotoxicity in vitro and improved therapeutic response in vivo. Combined BUB1 and CDK4/6 inhibition showed strong synergy (C.I. < 1) and enhanced radiosensitization in RB+ models. CDK4/6-resistant cells displayed increased BUB1 expression, and BUB1 inhibition partially restored sensitivity. Mechanistically, dual inhibition intensified homologous recombination defects, marked by persistent gH2AX and altered RAD51, RPA, and BrdU dynamics, consistent with sustained single stranded DNA and impaired HR repair. Persistent RPA32 Ser33 phosphorylation reflects ATR-dependent resection that requires BLM and EXO1 at later stages, supporting sustained resection and unresolved repair leading to increased cell death. Conclusions: Dual inhibition of BUB1 and CDK4/6 represents a promising therapeutic strategy for enhancing radiosensitivity in GBM, lung cancer, and TNBC, particularly in Rb-intact settings. Full article

Triple-negative breast cancer (TNBC) is an aggressive and heterogeneous subtype lacking effective targeted therapies. Its tumor microenvironment (TME) exhibits distinct features, including acidity, redox imbalance, elevated reactive oxygen species (ROS), and hypoxia, which provide exploitable triggers for targeted drug delivery. Stimuli-responsive polymeric nanocarriers have emerged as promising platforms that enable spatiotemporally controlled and site-specific therapeutic release in response to these endogenous cues, as well as exogenous stimuli such as temperature and light. These systems improve drug accumulation, penetration, and therapeutic efficacy while reducing systemic toxicity. Unlike previous reviews that broadly discuss nanocarriers in cancer therapy, this review focuses on the structure–function relationships of TME-responsive polymeric systems in TNBC and their translational limitations. We summarize recent advances in pH-, redox-, ROS-, hypoxia-, photo- and temperature-responsive polymers, highlighting their design strategies and therapeutic applications. Key challenges, including stimulus heterogeneity, limited in vivo validation, and clinical translation barriers, are also discussed. This review provides a concise framework for the rational design of programmable, multi-responsive polymeric nanomedicines for TNBC therapy. Full article

Purpose: Pembrolizumab combined with chemotherapy is the standard of care for early-stage triple-negative breast cancer (TNBC) following KEYNOTE-522. However, real-world data on immune-related adverse events (irAEs) remain limited. We evaluated the incidence, severity, and clinical consequences of irAEs in a real-world cohort and compared outcomes with KEYNOTE-522. Methods: We conducted a retrospective cohort study of patients with stage II–III TNBC treated according to the KEYNOTE-522 regimen at two Ontario cancer centres between June 2022 and May 2024. Data on irAEs, treatment discontinuation, and pathological complete response (pCR) were collected and contextualized against trial outcomes. Results: Among 79 patients, the pCR rate was higher than that reported in KEYNOTE-522 (77.2% vs. 64.8%). irAEs were documented in 51.9% of patients, compared with 33.5% in the trial. Permanent discontinuation due to irAEs occurred in 22.8% of patients, compared with 15.7% reported in KEYNOTE-522. Most discontinuations occurred during the neoadjuvant phase. Conclusions: Real-world patients experienced higher observed rates of low-grade irAEs and treatment discontinuation than reported in KEYNOTE-522; however, differences in study design limit direct comparison. These findings highlight the need for optimized toxicity management and further study of the impact of treatment duration on long-term outcomes. Full article

Adjuvant and perioperative immune checkpoint inhibitors (ICIs) have created a growing population of patients who relapse after prior programmed death-1 or programmed death-ligand 1 blockade, yet these patients were underrepresented in many trials that established metastatic standards. We performed a structured narrative review of PubMed/MEDLINE, ClinicalTrials.gov, reference lists, and international oncology guideline repositories through 15 August 2026. Eligible reports addressed recurrence patterns or treatment after curative-intent ICI in melanoma, non-small-cell lung cancer (NSCLC), renal cell carcinoma (RCC), urothelial carcinoma, or triple-negative breast cancer (TNBC); landmark metastatic studies were included only when direct evidence was unavailable and are labeled as extrapolation. Timing was standardized as on-treatment recurrence, early off-treatment recurrence (after the last ICI dose through 12 months), and late recurrence (>12 months). Shorter disease-free interval is consistently prognostic, but treatment-by-timing interactions are rarely available; timing should not be described as a validated pan-tumor predictive biomarker. Direct post-adjuvant evidence supports switching away from anti-PD-1 monotherapy for melanoma recurring on treatment, while selected late relapses may retain sensitivity. In RCC, retrospective post-adjuvant data support VEGF-targeted options, whereas CONTACT-03 and TiNivo-2 discourage routine ICI-TKI rechallenge specifically after prior ICI-treated metastatic RCC. Evidence in NSCLC, urothelial carcinoma, and TNBC is largely indirect. IMpassion132 was not an ICI-rechallenge trial, and only a small minority of ASCENT-04 participants had prior perioperative ICI. Treatment should integrate tumor-specific biology, actionable alterations, recurrence distribution, prior toxicity, comorbidity, access, and patient preference. Prospective trials dedicated to post-adjuvant ICI recurrence are needed. Full article

Background/Objectives: Triple-negative breast cancer (TNBC) exhibits substantial metabolic heterogeneity and plasticity, contributing to variable therapeutic responses. We investigated whether optical redox imaging (ORI) could characterize metabolic phenotypes, monitor responses to metabolic perturbation, and relate these responses to functional outcomes in TNBC cells. Methods: Four TNBC cell lines were treated with the lactate dehydrogenase A inhibitor FX11 or the glutaminase inhibitor CB-839, alone or in combination with paclitaxel. Intensity-based label-free ORI was performed and followed by imaging of fluorescent probes to assess mitochondrial membrane potential (MMP), re-active oxygen species (ROS), and cell number in the same dishes. Seahorse assays were used to evaluate mitochondrial respiration and glycolytic flux. Results: TNBC cell lines exhibited distinct basal redox phenotypes and differential responses to acute glycolytic and glutaminolytic perturbation. HCC1806 cells showed the strongest acute ORI responses to both FX11 and CB-839. Acute FX11 treatment induced a rapid reductive shift accompanied by ROS accumulation and loss of MMP, whereas CB-839 produced a more modest early reductive response without detectable ROS accumulation or MMP loss. Prolonged treatment revealed distinct temporal redox trajectories in HCC1806 cells: FX11-treated cells evolved from an acute reductive response toward a more oxidized state, while CB-839-treated cells transitioned from an early reductive shift to a sustained oxidized redox state accompanied by marked reductions in OCR and ECAR. Functionally, in two representative models (HCC1806 and MDA-MB-231), CB-839 reduced cell numbers and enhanced the anti-proliferative effect of paclitaxel, whereas FX11 had no significant effect on cell number despite inducing pronounced acute redox perturbations. Conclusions: Integrating ORI with metabolic flux assays and imaging-based functional measurements enables characterization of multiple dimensions of metabolic behavior, including basal phenotype, pathway-specific responsiveness, temporal redox responses, and treatment-associated outcomes. These findings support intensity-based wide-field ORI as a practical and accessible tool for probing metabolic heterogeneity and characterizing context-dependent metabolic responses in TNBC cells. Full article

Background/Objectives: Breast cancer (BC) is the most frequently diagnosed malignancy and a leading cause of cancer-related mortality in women worldwide. This disease is highly heterogeneous and dynamic, and chromosomal instability (CIN) plays a key role in the acquisition of these traits by generating genetic diversity that promotes tumor adaptation and influences therapeutic response and prognosis. Although several methods have been developed to quantify CIN, they are not readily applicable to human tumors and are limited in resolution, hindering a comprehensive understanding of intratumoral heterogeneity. In this study, we aimed to quantify CIN levels and clonal heterogeneity (CH) in HER2-positive (HER2+) and triple-negative (TNBC) breast cancer using single-cell RNA sequencing (scRNA-seq) data. Methods: We analyzed publicly available scRNA-seq data from HER2+ and TNBC tumors and non-malignant controls. CIN was scored at single-cell resolution using transcriptomic signatures, clonal heterogeneity was estimated from single-cell diversity metrics, and copy number alterations were inferred computationally. Differential expression and functional enrichment analyses were performed between cells with very low and extreme CIN levels, with key comparisons confirmed at the patient level. Results: Our analyses revealed pronounced intra- and intertumoral heterogeneity, with higher CIN levels in TNBC than in HER2+ and control samples. Genes differentially expressed in cells with extreme CIN values were mainly involved in cell division and related processes, and included candidate biomarkers not previously reported in this context. Our findings suggest a positive but statistically non-significant trend was observed between CIN and CH. Conclusions: Single-cell approaches such as scRNA-seq provide a powerful framework to elucidate CIN-related mechanisms and to identify potential biomarkers of BC aggressiveness and prognosis, supporting their further application in the study of intratumoral heterogeneity. Full article

Functional studies have shown that long noncoding RNAs (lncRNAs) play different roles in gene expression regulation, acting as epigenetic factors. Accumulating evidence indicates that lncRNAs modulate diverse biological processes, and their altered expression is associated with the development and progression of cancer, including BC. Here, aiming to identify lncRNAs associated with breast cancer, we performed a re-analysis of the expression data from 1071 tumors of eligible patients in the LACRN-MPBC Study. Using normalized data, we compared the expression profiles of triple-negative breast cancer (TNBC; 170 cases) and a heterogeneous group of non-TNBC tumors (782 cases). A total of 59 differentially expressed lncRNAs (DELncRNAs) could be identified based on the criteria of |log2FC| ≥ 1 and FDR < 0.05, using the DESeq2 R package. In the Kaplan–Meier survival analysis, we identified DELncRNAs that affect disease-free survival and/or overall survival, with eight of them in TNBC and 27 in non-TNBC patients. Multivariate Cox proportional-hazards models were used to evaluate independent predictors of survival. Two DELncRNAs, VLDLR-AS1 and PART1, were identified as independent prognostic markers for TNBC patients, while seven (LINC00239, FAM30A, LINC00842, LINC01315, EGOT, LY6E-DT, and SLC25A21-AS1) were independent prognostic markers for non-TNBC patients. Our computational study identifies several candidate lncRNAs associated with clinical outcomes in breast cancer. However, the DELncRNAs identified here should be interpreted as preliminary candidates, which require future validation and functional studies to determine their biological roles and evaluate their potential as prognostic biomarkers. Full article

Triple-negative breast cancer (TNBC) remains a major therapeutic challenge due to pronounced molecular heterogeneity, transcriptional plasticity, and frequent treatment resistance. MicroRNA (miRNA)-based strategies have emerged as potential approaches for modulating dysregulated gene expression networks in TNBC; however, the contribution of understudied miRNA families to TNBC-associated regulatory programs remains incompletely understood. This study aimed to investigate the tumor-suppressive role of miR-548f-3p in TNBC and to identify candidate downstream effectors, with particular focus on ANP32E. An integrative analysis combining public transcriptomic datasets, clinical expression profiling, computational target prediction, network-based prioritization, pathway analysis, and single-cell transcriptomic assessment identified miR-548f-3p as consistently downregulated in TNBC. Among candidate downstream targets, ANP32E, a chromatin-associated regulator involved in H2A.Z histone variant dynamics, was identified as a potential effector exhibiting increased expression in TNBC and enrichment within malignant epithelial cell populations. An inverse association between miR-548f-3p and ANP32E expression was observed in patient-derived samples. In breast cancer cell models, miR-548f-3p mimic restoration increased apoptosis, promoted G₀/G₁ accumulation, and reduced migration- and invasion-associated readouts, although measurable effects were also observed in non-tumorigenic MCF-10A cells. These phenotypic changes were accompanied by reduced ANP32E expression at the protein level, indicating that ANP32E expression is responsive to miR-548f-3p restoration. This study supports miR-548f-3p as a candidate tumor-suppressive miRNA in TNBC. The reduction in ANP32E protein expression following miR-548f-3p restoration, together with computational, single-cell, and clinical expression evidence, supports ANP32E as an expression-responsive candidate downstream effector of miR-548f-3p. Further reporter-based and rescue experiments are required to confirm direct 3′UTR-mediated targeting and to define the mechanistic contribution of ANP32E within the broader miR-548f-3p regulatory network. Full article

Background: Triple-negative breast cancer (TNBC) lacks effective targeted therapies, and its dependence on glycolysis represents a potential metabolic vulnerability. EIF3H, the largest subunit of the eukaryotic translation initiation factor 3 complex and a putative deubiquitinase, has been implicated in tumor progression, but its role in TNBC metabolism remains unclear. Methods: EIF3H expression and prognostic value were evaluated in public datasets, clinical TNBC specimens, and cell lines. Functional roles were examined using proliferation, colony formation, migration, and xenograft assays. Mechanisms were investigated by mass spectrometry, co-immunoprecipitation, ubiquitination and glycolytic rate assays, macrophage co-culture, and single-cell transcriptomic analysis. Results: EIF3H was significantly upregulated in TNBC and associated with poor survival. Transcriptionally activated by TRPS1, EIF3H bound to lactate dehydrogenase A (LDHA), reduced its ubiquitination, and prevented its proteasomal degradation. LDHA stabilization enhanced glycolysis and lactate production, thereby promoting TNBC cell proliferation and migration in vitro and tumor growth in vivo; these effects were abolished by LDHA knockdown and restored by LDHA re-expression. In addition, tumor-derived lactate induced M2 macrophage polarization via GPR65, which in turn reinforced malignant progression. Conclusions: Our findings define a TRPS1–EIF3H–LDHA axis that drives glycolysis-dependent TNBC progression and reveal lactate–GPR65 signaling as a mediator of tumor–macrophage crosstalk, supporting EIF3H as a potential prognostic biomarker and therapeutic target in TNBC. Full article