Circulating tumor DNA (ctDNA) has emerged as a clinically valuable biomarker for cancer detection, treatment monitoring, and minimal residual disease assessment. Despite its growing clinical utility, the biological mechanisms governing ctDNA release remain incompletely understood. The objective of this study was to investigate how chemotherapy-induced cytotoxicity and chemoresistance influence the kinetics and fragmentation patterns of cfDNA released by cancer cells in vitro.

Human lung adenocarcinoma (A549) and esophageal adenocarcinoma (FLO-1, OE19) cell lines were used to investigate cell-free DNA (cfDNA) release and fragmentation in vitro. Cancer cells were treated with chemotherapy (cisplatin and 5-fluorouracil), and cfDNA released into the culture medium was quantified as total cfDNA by Qubit fluorometry and mutation-specific cfDNA by droplet digital PCR (ddPCR). A cisplatin-resistant OE19 model was generated to directly compare cfDNA release kinetics between chemosensitive and chemoresistant cells. Fragment size distributions were determined and evaluated alongside cell death mechanisms assessed by flow cytometry.

cfDNA release correlated positively with viable tumor cell number across all cell lines. Chemotherapy increased per-cell cfDNA release in all cell lines. Chemosensitive OE19 cells exhibited significantly higher cfDNA release compared to chemoresistant OE19 cells following cisplatin exposure, with distinct temporal release kinetics. Additionally, chemotherapy treatment induced a shift toward the release of larger DNA fragments in both chemosensitive and resistant cells. This was accompanied by changes in PI-positive and Annexin V/PI double-positive cell populations, suggesting altered cell death processes following cisplatin treatment.

Chemotherapy-induced cytotoxicity significantly influences cfDNA release kinetics and fragmentation patterns, with distinct effects in chemosensitive and chemoresistant cancer cells. These findings provide mechanistic insight into tumor-derived DNA release biology and may have important implications for the interpretation, timing, and standardization of liquid biopsy testing during treatment. Furthermore, they establish in vitro cancer models as a valuable platform for studying cfDNA dynamics and informing preclinical therapeutic development.

Artificial intelligence (AI) and machine learning (ML) now reach into every stage of AML care. Deep learning models read therapy-relevant mutations directly from bone marrow smears; automated flow cytometry gating reproduces expert calls in under a minute; and the first AI pathology devices for hematology have cleared regulatory review and entered clinical use. Beyond diagnosis, ML captures the age-dependent weight of individual mutations that categorical ELN scoring misses, drug response prediction for venetoclax-azacitidine has been validated across multiple external cohorts, and large language models are being tested for tumor board support and trial matching. The next wave, from clonal architecture modeling and single-cell foundation models to digital twins and reinforcement learning for adaptive dosing, could move AML management from reactive toward predictive, evolution-aware care. This review departs from existing AI-in-hematology surveys in three ways: we (i) restrict the scope to AML and organize the field around clinical decision points rather than technology categories, (ii) grade every tool on a five-tier author-defined clinical readiness level (CRL-AML 1-5), which exposes hundreds of models clustered at CRL-AML 1-2 and none yet in prospective clinical evaluation, and (iii) close with a numbered three-year agenda naming the consortia, datasets, and pragmatic trials needed to carry the field from publication to practice.

Breast cancer relapse in bone is a significant clinical problem that is experienced in ~70-80% of patients with late-stage breast cancer. This condition commonly occurs 5-10+ years following surgical removal of the primary tumour. The long latency seen prior to relapse in bone is a result of tumour cell dormancy. Once disseminated to the bone, tumour cell interaction with the bone metastatic niche (endosteal niche and endovascular cells) maintains cells in a dormant state until changes to the local environment activate the niche to support outgrowth. Amassing evidence suggests that cytokines are key regulators of the bone metastatic niche, controlling bone homing and metastatic outgrowth. Pro-inflammatory cytokines, including IL-1β, IL-6, IL-8, TGFβ and RANKL, play crucial roles in attracting tumour cells to bone. Furthermore, these cytokines act in conjunction with IFN, VEGF, TGF, PTHrP, FGF, OPG and various chemokines to regulate expansion of the niche, facilitating tumour cell escape from dormancy and promoting the "vicious cycle of bone metastasis". Here, we review the current literature to provide an up-to-date understanding of how interactions between cytokine signalling cascades regulate the bone metastatic niches to promote homing, dormancy or metastatic outgrowth of breast cancers. Because breast cancers are predominantly osteolytic, this review focuses on dormancy and metastatic outgrowth associated with lytic disease in addition to current advances in novel therapeutics aimed at preventing this condition through targeting dormant cells.

Established oral squamous cell carcinoma (OSCC) remains constrained by local recurrence, inadequate lesion exposure, systemic toxicity, and therapy resistance. This narrative review evaluates a proposed microneedle-small extracellular vesicle (sEV) system for localized adjunctive treatment in three preclinical contexts: postoperative residual-disease control, local immunomodulation/checkpoint combination, and chemoradiotherapy sensitization. Direct evidence for the combined microneedle-sEV system in OSCC is not yet available; the analysis therefore integrates OSCC-specific sEV mechanisms with oral-mucosal microneedle and cross-disease engineering studies. We focus on whether the proposed microneedle-sEV system can preserve sEV potency, achieve reproducible local delivery, and meet oncologic-safety, manufacturing, and repeated-dose oral-safety requirements. The review provides a preclinical decision framework and comparator-based development criteria rather than a claim of clinical readiness.

The Antigen Processing and Presentation Machinery (APM) is essential for immune surveillance by enabling the presentation of antigenic peptides to T lymphocytes and facilitating the elimination of infected or transformed cells. In cancer, the integrity of this process influences cancer immune responsiveness (CIR), defined as a tumour's capacity to be recognised by the immune system and respond to immunotherapy. Tumours with intact antigen presentation pathways are more likely to generate effective antitumour responses, whereas APM defects promote immune escape and therapeutic resistance. Cancer cells frequently evade immune detection through altered antigen processing or reduced expression of major histocompatibility complex (MHC) class I molecules, limiting tumour antigen presentation to cytotoxic T lymphocytes. These alterations are increasingly recognised as determinants of response to immune checkpoint inhibitors and potential predictive biomarkers. APM defects may be reversible or irreversible. Interferon-mediated signalling can restore MHC class I expression and T-cell cytotoxicity in some tumours, whereas permanent genomic alterations affecting human leukocyte antigen (HLA) class I genes, β2-microglobulin (β2-m), or interferon-γ (IFN-γ) pathway components can severely impair antigen presentation. Emerging evidence highlights four mechanistic levels of APM perturbation: peptide generation, peptide loading, MHC class I integrity, and epigenetic regulation. Each contributes to distinct patterns of immune evasion. This review examines how MHC class I alterations influence CIR and contribute to immune evasion and immunotherapy resistance in gastrointestinal malignancies, while discussing therapeutic strategies to restore or bypass APM deficiencies.

Advances in anticancer therapies have substantially improved cancer survival but have also highlighted the growing challenge of cancer therapy-related cardiac dysfunction. Ferroptosis, an iron-dependent form of regulated cell death characterized by iron dysregulation, lipid peroxidation, and impaired antioxidant defense, has emerged as a promising strategy for eliminating therapy-resistant tumors. However, the lack of tissue specificity in ferroptosis regulation raises concerns regarding its potential contribution to cardiovascular injury during anticancer treatment. This review summarizes the dual roles of ferroptosis in cancer biology and cardio-oncology. We first discuss the molecular mechanisms governing ferroptosis, including iron metabolism, lipid peroxidation, and antioxidant defense systems. We then highlight the context-dependent roles of ferroptosis in tumor progression, immune regulation, and metabolic adaptation. Furthermore, we systematically review how chemotherapy, targeted therapy, immunotherapy, and radiotherapy contribute to ferroptosis-associated cardiovascular toxicity. Finally, we discuss emerging approaches to minimize cardiac injury, including tissue-specific ferroptosis-targeting and cardioprotective strategies. Understanding tissue-specific ferroptosis regulation may facilitate the development of safer and more precise therapeutic approaches in cardio-oncology.

Aptamer-functionalized liposomes are a promising strategy to improve the selectivity of anticancer therapies. AT11-L2 is a G-quadruplex (G4)-forming aptamer with high affinity for nucleolin (NCL), which is overexpressed at the surface of non-small cell lung cancer (NSCLC) cells. Here, we developed AT11-L2-functionalized liposomes loaded with doxorubicin (DOX) or BRACO-19 for targeted delivery to NSCLC cells. The effects of both compounds on AT11-L2 stability and the ability of the aptamer to retain G4 folding after liposome conjugation were evaluated. Liposomes were characterized for size, polydispersity, surface charge, stability, encapsulation efficiency, and release profile. Biological activity was assessed in A549 and MRC-5 cells. Liposomes had an average size of approximately 110 nm, with a slight size increase and reduced surface charge after AT11-L2 functionalization. The aptamer retained G4 folding in 100 mM KCl after conjugation. Encapsulation efficiency was approximately 90%. DOX showed substantial release within 72 h, whereas BRACO-19 exhibited a more sustained profile. AT11-L2-DOX liposomes showed preferential effects in A549 cells, while BRACO-19 formulations displayed lower selectivity and cytotoxicity. Additionally, the NCL-dependent internalization of AT11-L2-functionalized liposomes was supported by a protein-blocking assay using an anti-NCL antibody. These results support AT11-L2-functionalized liposomes as a versatile NCL-targeted delivery system for NSCLC.

Colorectal cancer (CRC) has shown significant heterogeneity regarding its response to immunotherapy. Long-lasting, beneficial effects have been observed in mismatch repair-deficient, microsatellite instability-high (dMMR/MSI-H) tumours, while mismatch repair-proficient, microsatellite stable (pMMR/MSS) tumours have remained resistant. Such differences in results have been studied in this review through the "hot" and "cold" tumour concept. It explains how various biological and microenvironmental factors play a role in immune resistance and T-cell priming and infiltration. Key factors include a low neoantigen load and defects in antigen presentation, which reduce the overall immune recognition of tumour cells. The review also studies certain processes such as Wnt/β-catenin and mitogen-activated protein kinase (MAPK) signalling and what input they have in the prevention of effective antitumour immune responses. Conventional treatments like chemotherapy and radiotherapy have been considered alongside more targeted treatments such as the inhibition of vascular endothelial growth factor (VEGF) signalling and the suppression of myeloid-mediated immune evasion, to convert "cold" MSS tumours into immune-responsive lesions. Methods which aim to modify the tumour microenvironment such as metabolic reprogramming and microbiome modulation have also been covered in this review. Artificial intelligence and nanomedicine are new technologies that could provide improved patient stratification and therapeutic precision, although their clinical application in pMMR/MSS CRC remains under investigation. A systematic literature search of PubMed and PubMed Central (PMC) was conducted from 10 June 2026 to 19 August 2026 using predefined eligibility criteria, with study selection reported according to PRISMA 2020. Because of substantial heterogeneity in study design, therapeutic approach and reported outcomes, the included evidence was synthesized narratively rather than by meta-analysis. A total of 158 studies were included.

Hydrogen is employed as a therapeutic gas in the treatment of stroke, tissue repair, and cancer, owing to its excellent biocompatibility, antioxidant properties, anti-inflammatory effects, and regulation of cellular metabolism. Moreover, nanometallic materials generate hydrogen through pH-responsive, light-responsive, ultrasound-responsive, and electrical stimulation and play a pivotal role in cancer therapy by activating anti-tumor immune responses, reversing immunosuppressive microenvironments, inducing immunogenic cell death, and sensitising radiotherapy and chemotherapy. Consequently, hydrogen therapy based on nanometallic materials has emerged as a novel research focus in cancer treatment. This review systematically elucidates the unique anti-cancer immunobiological effects of hydrogen therapy based on novel nanometallic materials. It meticulously analyses the reversal effects of different metals (Ca, Mg, Fe, Cu, Yb, etc.) in overcoming obstacles within the cancer immune cycle (including antigen presentation, T-cell activation, and resistance mechanisms). It highlights the structure-activity relationships between 'metal type-specific activity-immune effects' in the latest hydrogen therapies, elucidates the primary signaling pathways involved in hydrogen-mediated immune regulation, and systematically summarises breakthrough advances in how hydrogen therapy modulates immune responses against tumors. Building upon current cancer treatment trends, this review will synthesise key factors from clinical translation and immunological research perspectives to propose future directions for the field, addressing prevailing challenges.

Inula helenium L. (elecampane) is a traditional medicinal plant whose roots contain both hydrophilic polyphenols and lipophilic sesquiterpene lactones. However, the influence of the raw material origin and the extraction solvent on its combined bioactivity profile remains unclear. Moreover, although isolated sesquiterpene lactones from I. helenium have been extensively investigated for their anticancer properties, the bioactivity of whole root extracts against human melanoma cells has remained largely underexplored. This study aimed to systematically compare the phytochemical characteristics and biological activity of I. helenium root extracts obtained from three distinct raw material sources and three solvents of increasing polarity, and to identify the main factors associated with the observed bioactivity through multivariate integration. In this study, I. helenium roots from three distinct sources-commercially available dried roots, wild-collected roots, and cultivated roots collected from the Botanical Garden of Vilnius University-were extracted with three solvents of increasing polarity (n-hexane, ethyl acetate, and 70% ethanol). Each extract was characterized by total phenolic content, four antioxidant assays (DPPH, ABTS, FRAP, CUPRAC), UV-Vis fingerprinting, antimicrobial testing against Staphylococcus aureus, Klebsiella pneumoniae, and Candida albicans, and a CCK-8 assay on SK-MEL-28 human melanoma and HEK293 non-cancerous cells. Multivariate integration (PCA, HCA, Spearman rank correlation) identified solvent polarity as the dominant driver of both the phytochemical and the biological profile (PC1 and PC2 together explained 87.9% of the total variance). Ethanolic extracts combined the highest phenolic content (up to 38.6 mg GAE/g dry extract) and the strongest antioxidant capacity across all four assays, with maximum values of 177.3 (DPPH), 328.9 (ABTS), 174.1 (FRAP), and 541.6 (CUPRAC) µmol TE/g DE (p ≤ 0.004). In the preliminary antimicrobial screening, ethanolic extracts showed near-complete to complete inhibition of S. aureus and C. albicans at 25 mg/mL, with MIC₉₀ values as low as 12.5 mg/mL against S. aureus and 6.25 mg/mL against C. albicans, whereas K. pneumoniae showed only marginal growth inhibition (median values below 20% across the entire tested concentration range). Strikingly, n-hexane and ethyl acetate extracts-despite negligible antioxidant activity-produced concentration-dependent cytotoxic effects. Six of the nine extracts reduced SK-MEL-28 viability while sparing HEK293 cells, with the cultivated Botanical Garden n-hexane extract showing the greatest difference across the tested cell models (SI > 1.50, p = 0.0003). Targeted solvent selection thus enables a single plant material to yield two functionally distinct bioactive fractions: more polar extracts with pronounced antioxidant and antimicrobial activities and less polar extracts with preferential activity against SK-MEL-28 within the cell models tested.