Epstein-Barr virus (EBV) is a widespread gamma-herpesvirus associated with a number of malignancies, including nasopharyngeal carcinoma and gastric cancer (GC/EBV+). The primary oncogenic protein of EBV is latent membrane protein 1 (LMP1). Genetic variability of the LMP1 protein affects its oncogenic activity and clinical manifestations.

PurposeThis study aimed to evaluate the diagnostic success of percutaneous biopsies performed for suspected musculoskeletal tumors and to identify clinical, anatomical, and procedural factors associated with the need for repeat biopsy (re-biopsy). The secondary objective was to assess the impact of multidisciplinary team (MDT) evaluation on reducing unnecessary re-biopsy procedures.MethodsA total of 565 patients (331 bone, 234 soft tissue lesions) who underwent percutaneous biopsy for suspected musculoskeletal tumors between 2020 and 2024 were retrospectively analyzed. Diagnostic success was defined as a biopsy yielding a definitive histopathological diagnosis sufficient to guide treatment planning without additional tissue sampling. Predictive factors including lesion size, depth, location, and performing specialty were evaluated using univariate and multivariate logistic regression analyses.ResultsThe initial diagnostic yield was 65.5%. Multivariate analysis revealed that lesion width <2 cm (OR = 2.01; 95% CI, 1.19-3.39; p = 0.009) and biopsy performed by an interventional radiologist (compared to an orthopedic surgeon) (OR = 5.25; 95% CI, 2.83-9.74; p < 0.001) were independent predictors of re-biopsy recommendation. Among 72 patients recommended for re-biopsy, MDT evaluation averted the need for a second procedure in 38 cases (52.8%).ConclusionSmaller lesion size (<2 cm) and the medical specialty performing the biopsy were independent predictors of re-biopsy recommendation, with the latter likely reflecting systematic differences in case complexity and procedural environment rather than operator skill. Structured MDT evaluation significantly reduced unnecessary repeat procedures.

Osteosarcoma (OS) is an aggressive primary bone tumour with high metastatic potential. Current treatments including surgery and chemotherapy are limited by side effects and chemoresistance, underscoring the need for novel therapies. This study aimed to investigate the antitumor potential of resveratrol (RSV), a natural polyphenol, as a novel treatment for OS. The effects of RSV were evaluated in two osteosarcoma cell lines (SAOS-2 and U2-OS). A viability assay established 100 μM as the effective concentration, and hyperspectral imaging confirmed cellular uptake. Apoptosis was measured via caspase-3/7 activity and Annexin V/PI staining, while qRT-PCR assessed pro-apoptotic gene expression. Flow cytometry evaluated cell-cycle progression, and a wound-healing assay measured migration. Gene expression analyses (qRT-PCR) examined markers of cell adhesion, tumour progression and epithelial-mesenchymal transition. Finally, RSV's impact on the Wnt/β-catenin pathway was determined by quantifying nuclear β-catenin accumulation and the expression of its downstream oncogenic targets. RSV inhibited cell proliferation and induced apoptosis, increasing caspase-3/7 activity and modulating apoptotic gene expression. RSV also caused cell cycle arrest in S-phase. It reduced the cells' migration and altered the expression of cell adhesion and tumour progression genes, promoting a less invasive phenotype. Notably, RSV decreased nuclear β-catenin accumulation, downregulated oncogenic targets like c-Myc and MMPs, and upregulated E-cadherin while reducing vimentin levels, suggesting a reversal of epithelial-mesenchymal transition. These results suggest that RSV may offer a promising therapeutic approach for osteosarcoma, modulating key pathways involved in tumour progression, metastasis and chemoresistance. Further studies are required to assess its clinical applicability.

We report a case of hereditary bilateral retinoblastoma due to a de novo germline inversion on chromosome 13, resulting in disruption of the RB1 gene. The patient is a 22-month-old female who initially presented to the emergency room at 11 months of age with an erythematous left eye and leukocoria of the right eye. Computed tomography (CT) of the brain and orbits showed solid internal calcifications arising from the posterior globes concerning for bilateral retinoblastoma. Magnetic resonance imaging (MRI) of the brain and orbits confirmed bilateral retinoblastoma without associated pineal region or suprasellar mass. On initial examination under anesthesia, the right eye showed one tumor in the macula and two tumors in the inferior mid-periphery. Sub-retinal seeding extended to the inferior periphery. The left eye was enucleated and pathology showed leptomeningeal extension along the optic nerve extending to the surgical margin. The patient was treated on a non-protocol treatment plan with five cycles of vincristine, carboplatin, etoposide, cyclophosphamide, and weekly intraventricular topotecan via Ommaya reservoir, followed by autologous stem cell rescue. Tumor analysis showed loss of pRB protein expression by immunohistochemistry and methylation copy number profiling showed several segmental gains and losses, including focal loss of RB1 on 13q. A 123-gene cancer predisposition germline panel using genome and exome sequencing initially did not identify any RB1 single nucleotide variants or insertion/deletions. Subsequent constitutional chromosome analysis for RB1 showed a paracentric inversion between bands 13q14.2 and 13q31. Optical genome mapping (OGM) showed that the proximal breakpoint of the balanced inversion at 13q14.2 was within intron 17 of RB1, while the distal breakpoint at 13q31.3 did not interrupt any known genes of clinical significance. We review the various molecular techniques that aided in diagnosis of this patient and provide a summary of similar RB1-disrupting structural variants reported in the literature.

Hepatocellular carcinoma (HCC) is a malignant tumor worldwide with a high mortality rate and recurrence rate. Numerous miRNAs are being applied to the healing and prognosis of HCC. A prior investigation predicted that miR-5003-3p was linked to HCC, but the relevant molecular mechanisms were not clear.

NOL1/NOP2/SUN domain family member 2 (NSUN2), a member of the RNA methyltransferase family responsible for catalysing 5-methylcytosine (m5 C) modifications, has been increasingly recognized as a key regulatory factor in the initiation and progression of cancer. A growing body of evidence indicates that NSUN2 is aberrantly expressed in multiple malignancies, such as hepatocellular carcinoma, breast cancer, and gastric cancer, where its overexpression is frequently associated with unfavourable clinical outcomes, increased tumour aggressiveness, and resistance to therapy. Through its m5C modification activity, NSUN2 influences RNA stability, translational efficiency, and molecular interaction networks, thereby modulating critical oncogenic signalling pathways, including Wnt/β-catenin, PI3K/AKT, and epithelial-mesenchymal transition (EMT). Moreover, NSUN2 has been shown to interact with non-coding RNAs and epigenetic regulatory factors, contributing to the remodelling of the tumour microenvironment and facilitating immune evasion. Although NSUN2 is predominantly characterized by its tumour-promoting functions, emerging studies also suggest context-specific tumour-suppressive roles, highlighting its functional complexity in cancer biology. This review aims to summarize recent advances in understanding the molecular mechanisms underlying NSUN2 function, its clinical significance, and its potential as a biomarker or therapeutic target, while also discussing the challenges in translating these findings into clinical practice. A deeper understanding of NSUN2's diverse roles in carcinogenesis may provide novel insights into RNA epigenetics and inform the development of innovative strategies for cancer diagnosis and treatment.

When tumor cells colonize distant organs during metastasis, they interact extensively with surrounding cells. These interactions often change the behavior of surrounding cell populations which collectively induce a protumor microenvironment that permits tumor cell outgrowth into overt, clinically detectable metastatic disease. The lung is one of the most common sites of breast cancer metastasis. A chronic wound repair-related phenotype developed within the lung microenvironment during metastatic outgrowth in immunocompetent preclinical mouse models of breast cancer. This phenotype was characterized by an increased number and activation of lung type II alveolar epithelial (AT2) cells surrounding growing metastases. Metastatic outgrowth significantly changed AT2 gene expression, resulting in a modified secretome. AT2-derived secreted factors also promoted triple-negative breast cancer growth. AT2-secreted factors are regulated by the cyclic adenosine monophosphate response element-binding protein (CREB). Targeting CREB signaling with the phosphodiesterase 4 (PDE4) inhibitor roflumilast reduced AT2 breast cancer reciprocal interactions in vitro and metastatic outgrowth in vivo. Finally, AT2 cells adjacent to metastases in lungs from patients with metastatic breast cancer expressed higher PDE4B compared with AT2 cells in normal lungs.

Folate-conjugated PLGA-PEG nanocarriers have emerged as a promising strategy for targeted delivery of anticancer agents such as 9-bromo-noscapine (9-Br-Nos) for targeting of folate receptors, which are overexpressed in breast cancer cells. They were synthesized and characterized for their size, morphology, encapsulation efficiency, and drug release profile. In vitro cytotoxicity studies have revealed an enhancement in anticancer efficacy of nano-formulated drugs compared to free drugs. Enhanced cellular uptake, improved mitochondrial membrane potential, and increased reactive oxygen species (ROS) production were observed in treated cells. Additionally, the spheroid disintegration assay was performed to evaluate the effect of the nano-formulation on the three-dimensional (3D) tumor model. Western blot analysis revealed changes in the expression of apoptosis-related proteins, further supporting the enhanced therapeutic potential of the nanocarrier system. In vivo pharmacokinetic studies demonstrated improvement in bioavailability and prolonged circulation of the nano-encapsulated drug. This study underscores the potential of nanocarrier-based drug delivery systems in improving the therapeutic efficacy of anticancer agents.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies because of its typically late diagnosis and limited treatment options, with surgical resection being the primary intervention. Emerging studies have consistently reported associations between PDAC and metabolic dysfunctions, including obesity, chronic inflammation, and diabetes. In this study, we investigated the molecular interplay between PDAC-associated genes and metabolic disorder pathways.

Despite initial success, cancer therapies often fail due to the emergence of drug-resistant cells. In this study, we use a mathematical model to investigate how cancer evolves over time, specifically focusing on the state of the tumor when it recurs after treatment. We use a two-type birth-death process to capture the dynamics of both drug-sensitive and drug-resistant cells. Assuming resistant cells have equal fitness, we analyze the clonal diversity of drug-resistant cells at the time of cancer recurrence, which is defined as the first time the population size of drug-resistant cells exceeds a specified proportion of the initial population size of drug-sensitive cells. We examine two clonal diversity indices: the number of clones and the Simpson's Index. We calculate the expected values of these indices at the time of cancer recurrence. Additionally, we examine these two indices conditioned on early recurrence in the special case of a deterministically decaying sensitive population, with the aim of addressing the question of whether early recurrence is driven by a single mutation that generates an unusually large family of drug-resistant cells (corresponding to a low clonal diversity), or if it is due to the presence of an unusually large number of mutations causing drug resistance (corresponding to a high clonal diversity). Our findings, based on both indices, support the latter possibility. Furthermore, we demonstrate that the time of cancer recurrence can serve as a valuable indicator of clonal diversity, providing new insights into the evolutionary dynamics of recurrent cancers.

Giant Cell Lesions exhibit variable aggressive clinical behavior. Understanding the molecular mechanisms of these lesions can facilitate a more personalized and effective therapeutic approach.

Smoothelin, a cytoskeletal protein expressed in smooth muscle cells (SMCs), colocalizes with α-smooth muscle actin and is typically restricted to the cytoplasm. Interestingly, in malignant smooth muscle tumors (SMTs), particularly uterine leiomyosarcoma (LMS), aberrant nuclear localization of smoothelin has been observed, though the underlying mechanism remains unclear. To investigate, 99 uterine SMT cases were analyzed, comprising 26 LMS and 73 non-LMS (typical leiomyoma, cellular leiomyoma, mitotically active leiomyoma, and tumors of uncertain malignant potential). Histopathological and immunohistochemical evaluations included mitotic activity, Ki-67 index, and smoothelin expression. In vitro, two humans uterine LMS cell lines (SK-LMS-1, SK-UT-1) and a primary uterine SMC line under proliferative conditions were examined. Nuclear smoothelin expression was significantly more frequent in LMS compared with non-LMS. Within LMS tissues, both cytoplasmic and nuclear positivity were noted. Smoothelin-positive regions exhibited significantly higher mitotic activity, whereas Ki-67 labeling and nuclear size variation showed no association with localization. In cultured LMS cells, proliferation correlated with increased nuclear smoothelin positivity. These findings suggest that smoothelin is linked to proliferative capacity in SMTs, and its altered subcellular distribution may have potential utility in the pathological evaluation of SMTs.

Oncocytic adrenal neoplasms are rare adrenocortical tumors characterized by abundant eosinophilic cytoplasm due to massive mitochondrial accumulation. Their biological behavior is often difficult to predict, and various diagnostic systems-including the Lin-Weiss-Bisceglia system, the Helsinki score, and the reticulin algorithm-are used to assess their malignant potential. We report a case of an oncocytic adrenocortical carcinoma associated with Cushing's syndrome and hyperandrogenemia in a 34-year-old woman. Histologically, the tumor showed diffuse growth of eosinophilic cells with marked pleomorphism and focal capsular invasion. Immunohistochemistry confirmed adrenocortical origin and cortisol production, with a Ki-67 labeling index of 25%. Ultrastructural examination revealed densely packed mitochondria with lamellar and tubulovesicular cristae, accompanied by numerous whorled smooth endoplasmic reticulum formations, suggesting active remodeling of the endoplasmic reticulum. Whole-exome sequencing identified a pathogenic GNAS R201S mutation, together with copy number losses of ARID1A, CDKN2A, and ZNRF3 with widespread copy number alteration. Despite multiple adverse prognostic indicators, the patient has remained disease-free for over 5 years following adrenalectomy and adjuvant low-dose mitotane therapy. These findings suggest that GNAS activation may drive steroidogenesis while attenuating malignant progression, as demonstrated by integrated morphologic and genomic assessment in this case.

Intraoperative microwave ablation (MWA) can be used simultaneously with liver resection in patients with colorectal liver metastases (CLM) with curative intent. It is uncertain whether this treatment concept is limited by the number of intraoperative MWAs or the number of CLMs. This study was performed to investigate whether the number of CLMs and intraoperative MWAs is associated with overall survival (OS) and recurrence-free survival (RFS).

This review summarizes the latest advancements in electrochemical biosensors for the detection of circulating tumor cells (CTCs). As a core component of liquid biopsy, CTCs are highly valuable for tumor diagnosis and treatment. However, conventional detection methods often fail to meet clinical requirements due to limitations such as low sensitivity and heavy reliance on biomarkers. Electrochemical sensors address these challenges by immobilizing biorecognition elements on electrode interfaces, converting CTCs binding events into quantifiable electrical signals. This approach enables high-sensitivity detection (at the single-cell level), rapid response, and portability. The review systematically explores two key optimization strategies: (1) surface modification for antifouling (e.g., zwitterionic materials, polyethylene glycol layers) and (2) signal transduction strategies (e.g., aptamer-mediated nucleic acid amplification, nanomaterial-based signal amplification), which significantly enhance detection specificity and sensitivity. Furthermore, the review evaluates the clinical utility of this technology in early diagnosis and treatment monitoring, while also discussing challenges such as standardization and clinical translation. This review provides theoretical and technical insights for the development of systematically exploring next-generation high-performance liquid biopsy platforms for cancer.

Retinoblastoma (RB) is typically associated with highly penetrant pathogenic sequence variants (PSVs) in the RB1 gene; however, some families exhibit low penetrance RB (LPRB). We aimed to determine the penetrance rate and identify genetic and clinical characteristics of LPRB. To that end two cohorts were analyzed: 250 genetically confirmed LPRB cases identified through systematic literature review and 78 classical germline RB (CGRB) from three international centers- Thailand, Korea, and Israel. Penetrance rate was estimated as the proportion of affected individuals among RB1 PSV carriers. Multivariate models assessed parent-of-origin effects and predictors of penetrance. PSVs were annotated with Combined Annotation Dependent Depletion (CADD) scores and mapped to pRB structural domains. LPRB penetrance ranged from 50% (125/250, non-age-adjusted, CI [43.8%-56.2%]) to 64% (125/196, age-adjusted, CI [56.8%-70.2%]). Paternal inheritance of RB1 PSV was associated with a significantly increased risk of LPRB in offspring (OR = 6.24; P < 0.0001). Clinically, LPRB were significantly more likely than CGRB to present with unilateral disease (OR = 9.3, P < 0.0001), diagnosed at an older age (13 Vs 6.5 months, P = 0.01), and affect males (OR = 2.4, P = 0.03). LPRB-associated PSVs showed lower CADD scores (OR = 1.5; P = 0.0008), indicating lower predicted pathogenicity, and were enriched in pRB's N- or C-terminal domains (OR = 3.2; P = 0.007), consistent with hypomorphic effects. In conclusion, LPRB shows a 50-64% penetrance rate, more likely to be paternally inherited, have unilateral presentation, and associated with hypomorphic RB1 PSVs in the terminal pRB regions. These findings support retitling 'low penetrance RB' to 'medium penetrance RB'.

Cancer biology is a constantly evolving field of study due to the dynamic and complex nature of biological systems. The unique role of in vitro assays in cell biology has led to numerous transformations in our understanding of the functional and structural details of the cells and tissues that comprise these systems. However, the traditional monolayer assays have been reported to fall short of the in vivo physiology of cells. This has led to the development of 3D cell models, such as spheroids and organoids, that aim to recapitulate the intricate structural and functional behaviour of in vivo tumours. This review describes passive methods of spheroid formation (scaffold-free and scaffold-based) and the limitations that have driven the development of engineered active design methods to increase the physiological relevance of the model. Traditional assays need to be modified to evaluate these models, accounting for their architecture, density, and microenvironment gradients. Current developments in performing 3D cell assays include increased reagent concentration and incubation periods; however, many protocols still require single-cell analysis. We present a review of assay developments that maintain the spatial and contextual information that makes the 3D models physiologically relevant. Additionally, we introduce the advances in microscopy techniques that provide deeper visualisation of these models.

Background and objectives Diffuse large B-cell lymphoma presents a significant challenge due to its high rate of treatment failure in 40% of patients. In this study we screened microRNAs as biomarkers in chemotherapy non-responding patients, to allow their early prognostication. Methods In the exploratory phase, whole transcriptome microRNA profiling was conducted on 10 diffuse large B-cell lymphoma cases. Three patients achieved complete remission, while seven had refractory or relapsed disease. The differentially expressed miRNAs were validated in 41 retrospective, treatment-naive diffuse large B-cell lymphoma biopsies, including the original 10 cases. Additionally, 33 cases with paired biopsy and plasma samples were prospectively evaluated using qRT-PCR to correlate miRNA expression with clinical outcomes. Functional validation to identify downstream pathways was done by knocking down identified miRNAs in JM-1 cells by semi-quantitative proteomics. Results miR-193b-5p, miR-1307-5p, and miR-671-5p expression were downregulated in refractory/relapsed diffuse large B-cell lymphoma biopsies. Plasma miRNA levels did not reflect prognosis. In vitro proteomics showed their impact on key oncogenic pathways, revealing significant enrichment of replication and transcription-related proteins. Interpretation and conclusions The expression of miR-193b-5p, miR-1307-5p, and miR-671-5p miRNAs in diffuse large B-cell lymphoma tissues may serve as predictive biomarkers.

Background and objectives Ovarian carcinoma is one of the most lethal carcinomas among females. Its high prevalence and shorter 5-year survival rate is due to the fact that most of the cases are diagnosed at later stages. This highlights the importance of early diagnosis through reliable biomarkers. We studied the diagnostic role of SOX9 protein in ovarian carcinoma and its diagnostic ability. The primary objective was to compare the level and clinical relevance of SOX9 protein in the tissues of patients with ovarian carcinoma with non-malignant ovarian tissues. Methods Tissue levels of SOX9 protein were estimated in the study and control groups (60 each group). SOX9 levels were compared between the study vs. control groups and also between high grade and low-grade ovarian cancer. SK-OV3 ovarian adenocarcinoma cell line was used as supportive evidence to prove the presence of SOX9 in malignant ovarian cells. Results Levels of SOX 9 protein (3.9±2.7 ng/mL) were high in tissue of ovarian cancer patients when compared to non-malignant (1.5 ±1.1 ng/mL) ovarian tissues. Higher levels of SOX 9 protein were found in tissues of ovarian cancer patients when compared to non-malignant ovarian tissues. The mean of SOX 9 levels in tissues of high-grade serous carcinoma was 3.5±2.5 ng/mL as compared to 1.0±0.9 ng/mL in low-grade serous carcinoma. Interpretation and conclusions SOX9 appears to be an important player in the molecular tumourigenesis of ovarian cancer, particularly in high grade tumours.

Background and objectives Radiotherapy for advanced cervical cancer (CaCx) often results in unintended genitourinary toxicities, notably bladder damage. Predicting such radiation-induced toxicity remains challenging. γ-H2AX, a marker of DNA double-strand breaks (DSBs), offers promise as a predictive biomarker for radiosensitivity. This study aimed to evaluate γ-H2AX foci kinetics in peripheral blood lymphocytes (PBLs) as a surrogate for DNA damage response and a predictor of bladder toxicity in CaCx patients undergoing pelvic radiotherapy. Methods In this prospective study, 43 FIGO stage IIIB CaCx patients were enrolled. Stage I (n=31) assessed γ-H2AX induction post-CT simulation (2-6 mGy); Stage II (n=34) evaluated γ-H2AX kinetics across three radiotherapy fractions (FR1, FR13, FR25) during external beam radiotherapy (50 Gy in 25 fractions ± cisplatin). Blood samples were collected at baseline, 1-, 4-, and 24-h post-irradiation. γ-H2AX foci were quantified via flow cytometry. Bladder toxicity was graded usingRadiation Therapy Oncology Group (RTOG) criteria. Results CT and radiotherapy both induced significant γ-H2AX foci, peaking at 1 h. Patients without bladder toxicity showed higher foci induction and faster decay (1→4h: 48.9% vs. 39.4%; 1→24h: 43.6% vs. 12.8%) across all fractions. Persistent foci at 24 h correlated with increased toxicity risk, indicating deficient DNA repair capacity. Interpretation and conclusions γ-H2AX foci kinetics effectively reflect in vivo DNA repair efficiency and predict radiation-induced bladder toxicity. This minimally invasive biomarker may guide personalized radiotherapy, enabling early identification of high-risk patients and potential use of radioprotectors or treatment modifications.