The aim of this study was to clarify the association between programmed death-ligand 1 (PD-L1) expression and the treatment efficacy of anaplastic lymphoma kinase (ALK)-tyrosine kinase inhibitors (TKIs), particularly alectinib, in patients with advanced or recurrent ALK-positive non-small cell lung cancer (NSCLC). This multicenter retrospective cohort study involved 129 patients with advanced or recurrent ALK-positive NSCLC treated between July 2014 and December 2022 at 13 Japanese institutions. All patients had available PD-L1 tumor proportion score (TPS) data evaluated using the 22C3 antibody and received at least one ALK-TKI. The primary endpoint was progression-free survival (PFS) with the first ALK-TKI. The secondary endpoints were overall survival (OS), objective response rate (ORR), and incidence of grade ≥ 3 adverse events (AEs). The low- (< 50%, n = 74) and high-PD-L1 groups (≥ 50%, n = 55) showed comparable ORRs (86.5%, 95% confidence interval [CI]: 76.5%-93.3% vs. 85.5%, 95% CI: 73.3%-93.5%; p = 1.00), median PFS (29.7, 95% CI: 18.8-51.2 months vs. 35.0, 95% CI: 16.6-54.2 months; hazard ratio [HR]: 1.01, 95% CI: 0.63-1.63; p = 0.961), and median OS (119.9, 95% CI: 60.1 months to NA vs. 76.6, 95% CI: 43.0 months to NA; HR: 1.74, 95% CI: 0.91-3.32; p = 0.095). PD-L1 expression was not significantly associated with survival. However, severe (grade ≥ 3) AEs were more common in the high-PD-L1 group than in the low-PD-L1 group (21.8% vs. 5.4%). No patients received concomitant cytotoxic chemotherapy or immune checkpoint inhibitors during treatment with the first ALK-TKI, and the frequency of prior systemic therapy before the first ALK-TKI was similar between the two PD-L1 groups. Intratumoral PD-L1 expression was not significantly associated with the efficacy of ALK-TKI therapy in this cohort of patients with ALK-positive NSCLC, particularly those treated with alectinib. The numerical OS difference and imbalance in grade ≥ 3 AEs should be interpreted cautiously because of the retrospective design and limited number of OS events. Further prospective studies with larger cohorts are warranted to validate these observations.

Dysregulation of ubiquitination drives tumor progression in various cancers, including clear cell renal cell carcinoma (ccRCC). RING finger protein 152 (RNF152) is a ubiquitin E3 ligase that regulates cancer development. This study aimed to investigate the role of RNF152 in ccRCC and to elucidate the underlying molecular mechanism. RNF152 expression and its correlation with survival were analyzed through bioinformatics analysis of the RNA expression profile from TCGA. Tumor cell viability and colony formation, as well as cytotoxicity of co-cultured CD8+ T cells, were assessed. Tumor growth and immune evasion were evaluated using a tumor-bearing mouse model. Furthermore, whether RNF152 regulates PD-L1 via ubiquitination was investigated. RNF152 was expressed at low levels in ccRCC, and high RNF152 expression correlated with better patient survival. Overexpression of RNF152 inhibited proliferation and immune evasion of ccRCC cells in vitro, while knockdown of RNF152 had the opposite effects. Moreover, RNF152 promoted degradation of PD-L1 by facilitating K48-linked polyubiquitination, and the ubiquitination site was identified as K64. Additionally, overexpression of PD-L1 counteracted the inhibition of proliferation and immune evasion induced by RNF152. In vivo, anti-PD-L1 therapy inhibited tumor growth and immune evasion in mice with RNF152 knockdown. facilitating ubiquitination of PD-L1. These findings reveal a novel regulatory mechanism in ccRCC and highlight the therapeutic potential of upregulating RNF152 to enhance anti-tumor immunity, potentially in combination with anti-PD-L1 immunotherapy.

Head and neck squamous cell carcinoma (HNSCC) remains a challenge due to recurrence, metastasis, and resistance to conventional therapies, particularly in human papillomavirus (HPV)-negative tumors. Cannabidiol (CBD), a non-psychoactive phytocannabinoid, has demonstrated antitumor activity in several cancer models; however, its effects in chemoresistant HNSCC are not well established. This study evaluates the antitumor effects of CBD and cellular mechanisms in a cisplatin -resistant, HPV-negative HNSCC model. Cytotoxic effects of CBD were assessed in HN13 cells and compared with fibroblasts and keratinocytes using MTT and selectivity index analysis. Migration, invasion, metalloproteinase activity, clonogenic survival, and cytotoxicity on 3D spheroids were evaluated. Cell death-related mechanisms were investigated using acridine orange/propidium iodide staining and Western blot analysis of apoptosis, autophagy, and cell-cycle regulation. Antitumor activity and vascular safety were further assessed using the chorioallantoic membrane model and HET-CAM assay. The combined effect of CBD and CIS was evaluated using the combination index analysis. CBD exhibited selective cytotoxicity toward HN13 cells and reduced migration, invasion, matrix metalloproteinase-2/9 activity, clonogenic survival, and 3D spheroid growth. CBD altered proteins associated with apoptosis, autophagy, and cell-cycle regulation. Pharmacological inhibition with 3-MA enhanced CBD-associated apoptotic signaling, increasing PARP and caspase-3 cleavage. In the CAM, CBD reduced tumor growth and vascular area without inducing vascular irritation. Furthermore, CBD and CIS demonstrated synergistic cytotoxic effects in HN13 cells. CBD exerted selective antitumor effects on CIS-resistant HN13 HNSCC cells, affecting tumor-associated behaviors and modulating apoptosis and autophagy-related signaling, while acting synergistically with cisplatin. These findings support further preclinical investigation of CBD as a potential adjunct approach in HNSCC, particularly in chemoresistant settings.

Although lipopolysaccharide (LPS) is increasingly implicated in inflammation, its role in tumor pathogenesis remains poorly characterized. C-X-C chemokine receptor type 4 (CXCR4) is markedly overexpressed in multiple malignancies, including gallbladder cancer (GBC). This study investigated the role of LPS in increasing CXCR4 expression and function and elucidated the underlying signaling mechanisms in GBC. Biliary LPS concentrations were quantified using a chromogenic Tachypleus amebocyte lysate assay. GBC cell migration and invasion were assessed using the wound-healing assay and Transwell chambers. CXCR4 expression profiles were determined through immunohistochemistry (IHC), quantitative real-time PCR (qRT‒PCR), Western blot analysis, and luciferase reporter assays. Elevated LPS levels in bile samples from GBC patients correlated with an advanced pT stage. Notably, LPS concentrations were positively associated with CXCR4 expression. In vitro, stimulation with LPS robustly increased the migratory and invasive capacities of GBC cells, which was mediated by the upregulation of CXCR4 expression. Mechanistically, the transcription factor nuclear factor-κB (NF-κB) bound to the CXCR4 promoter to drive its expression, and LPS potentiated this transcriptional activation. Our findings reveal a novel LPS/NF-κB/CXCR4 signaling axis that may contribute to the aggressive phenotype of GBC in vitro, providing a potential mechanistic link between chronic biliary inflammation and tumor progression. Further in vivo studies and clinical validation are warranted.

The mitochondrial solute carrier (SLC25) family regulates solute transport across the mitochondrial inner membrane. Among them, SLC25A39 has emerged as a critical player in tumor biology, primarily through its dual role in mediating mitochondrial glutathione (mGSH) transport and maintaining iron-sulfur cluster homeostasis. Recent evidence implies that SLC25A39 dysfunction disrupts mGSH metabolism, thereby influencing iron-sulfur cluster synthesis and driving metabolic reprogramming in tumors. Notably, SLC25A39 is governed by a GSH-Fe-S-AFG3L2 feedback circuit, a feature not shared by its closest homolog SLC25A40. Additionally, upregulation of SLC25A39 is frequently observed in multiple malignancies, including colorectal, pancreatic, and liver cancers, where it correlates with enhanced proliferation, metastasis, and poor prognosis. Furthermore, SLC25A39 expression is intricately associated with remodeling of the tumor immune microenvironment and the development of therapeutic resistance, although these associations are primarily based on correlative evidence and require functional validation before causal conclusions can be drawn. This review synthesizes current knowledge on the molecular mechanisms by which SLC25A39 integrates mGSH transport with iron metabolism, and critically evaluates its emerging roles across different cancer types. We propose that beyond serving as a candidate prognostic biomarker, SLC25A39 represents a potential therapeutic target whose clinical utility remains to be established, and we discuss key unanswered questions regarding its tissue-specific functions and the translational challenges that lie ahead.

Ginger (Zingiber officinale) is a common spice with therapeutic potential due to its bioactive compounds and was used for the green synthesis of ginger-derived silver nanoparticles (G-AgNPs). The cytotoxic, antioxidant, and pro-apoptotic effects of the synthesized G-AgNPs were evaluated against HepG-2, MCF-7, and WI-38 cell lines. Gas chromatography-mass spectrometry (GC-MS) analysis identified 36 phytochemicals, with major components including α-zingiberene (37.88%), β-sesquiphellandrene (15.65%), 6-gingerol (10.68%), α-farnesene (8.04%), β-bisabolene (7.83%), α-curcumene (6.96%), 8-gingerol (0.96%), and 6-shogaol (0.95%). Characterization of G-AgNPs using ultraviolet-visible spectroscopy (UV-Vis), Fourier-transform infrared spectroscopy (FT-IR), zeta-potential analysis (- 46.17 ± 1.42 mV), and transmission electron microscopy (TEM) revealed spherical nanoparticles with an average diameter of 10.44 nm. Cytotoxicity assays demonstrated greater inhibition of HepG-2 and MCF-7 cell viability than of WI-38 cells, with IC₅₀ values of 1.20 ± 0.03 µM (4.51 ± 0.11 µg/mL) and 1.49 ± 0.05 µM (5.60 ± 0.19 µg/mL), respectively, compared with 2.49 ± 0.07 µM for WI-38 cells. Treatment with G-AgNPs significantly increased caspase-3-associated protein levels in both HepG-2 (13.81 ± 0.35 ng/mg protein) and MCF-7 (42.83 ± 1.47 ng/mg protein), and decreased Bcl-2 expression (HepG-2: 2.12 ± 0.14 ng/mg protein; MCF-7: 3.17 ± 0.03 ng/mg protein). However, given the known deficiency of functional caspase-3 in MCF-7 cells, the observed increase likely reflects caspase-3-immunoreactive protein rather than active enzyme. Antioxidant evaluation demonstrated free radical scavenging activity (IC₅₀ = 28.96 µg/mL), comparable to ascorbic acid (IC₅₀ = 14.03 µg/mL), along with significant CT- DNA intercalation. In vivo, treatment of Ehrlich ascites carcinoma (EAC)-bearing mice with G-AgNPs significantly reduced abdominal circumference, ascitic fluid volume, and viable tumor cell counts while increasing the percentage increase in lifespan by 93.3%. G-AgNPs also improved liver and kidney function, increased hepatic GSH, SOD, TAC, and catalase levels relative to untreated EAC-bearing mice, reduced oxidative stress markers (MDA and NO), and ameliorated liver histopathological alterations, with fewer adverse effects than cisplatin. The findings indicate that G-AgNPs possess potential anticancer activity and preliminary differential cytotoxicity toward HepG2 cells compared with non-cancerous cells. Further toxicological and in vivo investigations are required to establish their safety and therapeutic applicability.

Cancer-associated fibroblasts (CAFs) are potential contributors to tumor progression and immune resistance in endometrial cancer (EC), but the extent of EC-specific mechanistic and therapeutic evidence remains uneven. Studies using EC-derived fibroblasts support tumor-promoting paracrine activity and provide experimental evidence of altered natural killer cell function. Single-cell and spatial analyses further characterize stromal heterogeneity and candidate intercellular interactions. However, many proposed mechanisms of CAF-mediated immune exclusion and most CAF-directed therapeutic strategies are informed by studies in other malignancies. This review distinguishes findings obtained in EC patient samples and experimental models from evidence extrapolated from pancreatic, breast, gastric, and other cancers. We examine CAF heterogeneity, soluble signaling, matrix remodeling, and immune-cell interactions, while identifying where functional validation in EC is still required. Therapeutic approaches, including FAP-directed interventions, stromal reprogramming, and signaling blockade combined with immune checkpoint inhibition, are discussed as candidate research strategies rather than established EC treatments. Priorities include validating CAF-immune mechanisms in EC, developing reproducible stromal biomarkers, and testing treatment combinations in appropriate disease-specific models and clinical studies.

Glioblastoma (GBM) is the most common malignant tumor of the adult central nervous system and is associated with an extremely poor prognosis. Despite standard-of-care treatment, including surgery, radiotherapy, and chemotherapy, mortality rates remain high, highlighting the urgent need for more effective therapeutic strategies. The pronounced intratumoral heterogeneity, highly invasive growth behavior, and strong dependence on the tumor microenvironment complicate both the biological understanding of glioblastoma and the management of therapy resistance. Conventional two-dimensional culture systems and simplified three-dimensional models fail to adequately capture these complex disease characteristics. Recent advances in three-dimensional organoid and assembloid technologies provide physiologically relevant platforms that preserve patient-specific molecular profiles, spatial organization, and tumor-microenvironment interactions. In particular, assembloid systems integrating multiple cellular components enable the investigation of glioblastoma invasion, microenvironment-driven cellular plasticity, and mechanisms of therapeutic resistance. This review provides a critical overview of current two- and three-dimensional in vitro models used in glioblastoma research, with a focus on emerging organoid and assembloid platforms. Their strengths, limitations, and recent innovations are discussed, highlighting their potential to advance translational research and support personalized therapeutic strategies for glioblastoma.

Oral squamous cell carcinoma (OSCC) develops within a highly regenerative mucosa but progressively remodels its surrounding tissue to support invasion, immune evasion, recurrence, and therapeutic adaptation. This review conceptualizes OSCC as a chronic, non-resolving wound-like state in which physiological tissue-repair programs become maladaptively co-opted. Tumor-driven remodeling involves extracellular-matrix (ECM) degradation, laminin reorganization, LOX-mediated matrix stiffening, cancer-associated fibroblast (CAF) heterogeneity, vascular alterations, and immune-stromal interactions at spatially defined niches such as invasive fronts. Surgical resection subsequently imposes acute tissue injury and wound-healing responses that can modify platelet, inflammatory, fibroblast, immune, and ECM programs within the surgical bed and histologically defined margins. Radiotherapy and chemotherapy introduce additional, non-equivalent remodeling trajectories involving oxidative stress, DNA damage, endothelial injury, fibrosis, matrix turnover, hypoxia, and immune modulation. Immune checkpoint inhibition acts upon this pre-existing and treatment-conditioned microenvironment, with heterogeneous CAF, myeloid, endothelial, and lymphoid organization influencing therapeutic response or resistance. Across these contexts, stromal remodeling is therefore dynamic rather than a static feature of the tumor microenvironment, and treatment-associated changes cannot always be distinguished from pre-existing tumor-driven alterations. We propose that this represents the "dark side" of tissue repair, whereby processes essential for tissue restoration may become permissive to tumor persistence and treatment adaptation. Integrating spatially resolved stromal and immune features with established clinicopathological factors may improve recurrence-risk stratification and therapeutic selection. Longitudinal studies are needed to distinguish transient repair responses from persistent tumor-permissive remodeling and to identify actionable stromal states in OSCC.

Colorectal cancer (CRC) remains a major global health burden, with incidence and mortality rates continuing to rise worldwide. By 2040, CRC cases are projected to reach 3.2 million, driven by complex interactions between genetic, environmental, and lifestyle factors. While the molecular mechanisms of CRC have been extensively studied, increasing evidence highlights the critical role of non-coding RNAs (ncRNAs), including microRNAs (miRNAs), long non-coding RNAs (lncRNAs), and circular RNAs (circRNAs) in regulating tumor progression, immune evasion, and therapeutic resistance. Recent research reveals that ncRNAs not only influence tumor cell behavior but also modulate the tumor microenvironment (TME), particularly by regulating tumor-associated macrophage (TAM) polarization. The balance between pro-inflammatory M1 and pro-tumorigenic M2 macrophages profoundly affects CRC progression and treatment response. Dysregulated ncRNAs, such as miR-934, lncRNA RPPH1, and circPLCE1, promote M2 polarization and metastasis via PI3K/AKT, NF-κB, and STAT3 signaling pathways, while others like miR-195-5p and lncRNA GAS5 favor M1 polarization and inhibit tumor growth. These molecules act through diverse mechanisms, including exosomal transfer, ceRNA interactions, and transcriptional regulation, forming intricate communication networks between CRC cells and immune components. Understanding these ncRNA-mediated pathways provides novel opportunities for therapeutic intervention, enhancing immunotherapy efficacy, and improving diagnostic precision through circulating exosomal ncRNA biomarkers. Collectively, targeting ncRNA-regulated macrophage polarization holds promise for developing next-generation precision therapies and advancing immunomodulatory strategies in CRC.

GFPT1, the rate-limiting enzyme of the hexosamine biosynthesis pathway, promotes various cancers but its role in esophageal cancer is unclear. GFPT1 expression was assessed by public datasets, western blotting, and IHC. GFPT1-silenced cells were established via siRNA. Proliferation, migration, invasion, apoptosis, ER stress, and autophagy markers were evaluated by CCK-8, colony formation, Transwell, wound healing, flow cytometry, western blotting, and LC3 immunofluorescence. In addition, rescue experiments were performed using the ER stress inhibitor 4-PBA in GFPT1-knockdown cells, and O-GlcNAcylation levels were examined by western blot. GFPT1 was highly expressed in esophageal cancer tissues. Knockdown of GFPT1 suppressed proliferation, invasion, and migration, while enhancing apoptosis. GFPT1 depletion upregulated ER stress markers (GRP78, GRP94, p-PERK, p-eIF2α) and autophagy-related changes (increased LC3-II/LC3-I ratio, ATG5, ATG7, and reduced p62), and reduced overall O-GlcNAcylation. Importantly, treatment with 4-PBA significantly reversed the GFPT1 knockdown-induced suppression of proliferation, migration, and invasion, and rescued the alterations in ER stress markers, indicating that the phenotypic effects are causally mediated, at least in part, through the ER stress pathway. GFPT1 knockdown impedes esophageal cancer progression by reducing aggressive phenotypes and promoting apoptosis, potentially via ER stress and autophagy activation.

Itaconic acid is an immunometabolite that regulates macrophage polarization within the tumor microenvironment and influences tumor progression and invasion. Although its anti-cancer and tumor-promoting effects remain controversial, its role in modulating macrophage-derived exosomes and their impact on breast cancer cells is not well understood. This study investigated the effects of exosomes derived from dimethyl itaconate (DMI)-treated macrophages on epithelial-mesenchymal transition (EMT), apoptosis, migration, and EMT-related miRNAs in MCF7 breast cancer cells. U937 macrophages were treated with 250 µM DMI for 48 h. Exosomes (DMI-EXO) were subsequently isolated, characterized, and used to treat MCF7 cells. The expression of EMT-related proteins (E-cadherin, N-cadherin, ZEB1/2, Slug, and Vimentin) and EMT-associated miRNAs (miR200C-5p, miR30C-5p, miR-638, and miR-137) was evaluated by Western blotting and quantitative real-time PCR, respectively. Apoptosis, cell migration, and invasion were assessed using Annexin V/PI staining, the scratch wound-healing assay, and the Transwell invasion assay, respectively. DMI-EXO treatment significantly increased the expression of the mesenchymal markers N-cadherin, ZEB1/2, Slug, and Vimentin while reducing the epithelial marker E-cadherin. In addition, the expression of EMT-suppressive miRNAs, including miR200C-5p and miR30C-5p, was significantly decreased, whereas no significant change was observed in miR-137 expression following DMI-EXO treatment. DMI-EXO treatment did not significantly alter apoptosis in MCF-7 cells but enhanced cell migration and invasion, as demonstrated by accelerated wound closure in the scratch assay and increased invaded area in the Transwell invasion assay. These findings indicate that DMI-induced macrophage-derived exosomes promote EMT and migration in breast cancer cells by modulating EMT-related proteins and miRNAs. This study provides new evidence that itaconic acid may facilitate breast tumor progression through exosome-mediated communication between macrophages and cancer cells.

Although papillary thyroid carcinoma (PTC) usually follows an indolent course, lymph node metastasis and locally advanced disease remain clinically important challenges. This study evaluated interleukin-1 receptor accessory protein (IL1RAP) in PTC using pan-cancer datasets, TCGA/GEO thyroid cancer cohorts, and in vitro validation. IL1RAP was upregulated in PTC tissues and cell lines, and higher IL1RAP expression was associated with advanced pathologic stage, lymph node metastasis, extrathyroidal extension, and unfavorable prognostic indicators. Functional assays showed that IL1RAP overexpression increased PTC cell proliferation, migration, invasion, and EMT-like changes, whereas transient IL1RAP silencing produced the opposite pattern. Bioinformatic enrichment and western blot analyses indicated that these cellular effects were accompanied by increased p-AKT with minimal change in total AKT, suggesting possible involvement of PI3K/AKT-related signaling. To our knowledge, this is the first systematic analysis linking IL1RAP to EMT-associated aggressive behavior in PTC. IL1RAP may represent a candidate biomarker associated with aggressive clinicopathological features, while its diagnostic/prognostic utility and therapeutic potential require further validation.

Ehrlich carcinoma (EC) and its ascitic form, Ehrlich ascites carcinoma (EAC), are among the oldest and most widely used experimental tumor models in cancer research. Although they have traditionally been used for anticancer drug screening, their potential to address broader aspects of aggressive tumor biology has received comparatively less attention. EC/EAC exhibits several characteristics associated with advanced malignancy, including rapid tumor growth, angiogenesis, immune dysregulation, metabolic changes, therapeutic resistance, and complex interactions between the tumor and host. These features make EC/EAC a useful complementary model for studying different aspects of tumor progression. However, research using these models has largely focused on transcriptional changes and individual signalling pathways, while post-transcriptional regulatory mechanisms remain less explored. In particular, the role of the RNA-induced silencing complex (RISC), a major component of microRNA-mediated gene regulation, is still poorly understood in EC/EAC. In this review, we discuss the major limitations of current EC/EAC research, including the limited use of multi-omics approaches, single-cell and spatial analyses, functional genomics, immune checkpoint profiling, and integrated systems-level studies. We further discuss how incorporating RISC-centered regulatory networks with emerging approaches such as CRISPR-based studies, spatial transcriptomics, and metabolic profiling could provide deeper mechanistic insights and improve the translational value of these models. Bringing together the strengths of this established tumor model with contemporary approaches in cancer biology could help reposition EC/EAC as a relevant and versatile platform for studying tumor progression and therapeutic responses.