Search Results (1,843)

Background/Objectives: OS is characterized by marked tumor heterogeneity, stemness-associated phenotypes, metastatic potential, and poor prognosis. FZD4 is an important Wnt signaling receptor involved in stem cell regulation and tumor progression, but its role and microRNA-mediated regulation in OS remain unclear. The aim of this study was to investigate the relationship between FZD4 expression, OS stemness, malignant progression, and upstream microRNA regulation. Methods:Human OS single-cell RNA sequencing data were analyzed using Seurat, CytoTRACE, and Monocle2. Tumor cell subpopulations, developmental potential, and pseudotime trajectories were evaluated. Functional enrichment analyses were performed to identify pathways associated with high FZD4 expression. MicroRNA–mRNA interaction analysis was used to predict upstream regulatory microRNAs, and immunohistochemistry, Western blotting, and in vitro and in vivo experiments were performed for validation. Results: FZD4 was mainly enriched in OS tumor cell subpopulations and was significantly associated with poor prognosis. Tumor cells with high FZD4 expression showed increased developmental potential, stemness features, and enrichment in early developmental states. Functional enrichment analysis indicated activation of epithelial–mesenchymal transition, angiogenesis, inflammatory response, KRAS signaling, hypoxia, glycolysis, and Wnt-related pathways. Clinical validation confirmed elevated expression of FZD4, OCT4, and SOX2 in OS tissues. FZD4 knockdown inhibited OS cell stem-like phenotypes and malignant behaviors, proliferation, and migration, while miR-424-5p suppressed these malignant phenotypes by targeting FZD4. Importantly, rescue experiments demonstrated that restoration of FZD4 substantially reversed the inhibitory effects of miR-424-5p mimics on sphere formation, migration/invasion, proliferation, and stemness-associated cell populations. Conclusions: Our findings identify miR-424-5p as an upstream suppressor of FZD4 and demonstrate that the miR-424-5p/FZD4 axis regulates stem-like and malignant phenotypes of OS cells, at least partly through Wnt/β-catenin signaling. This axis may represent a potential therapeutic target for OS. Full article

Background: Optimal porosity and surface area of additively manufactured Ti-6Al-4V scaffolds for early human mesenchymal stem cell (hMSC) retention remain unclear, despite their importance in reconstruction of critical-sized bone defects. This study evaluated scaffold architectures fulfilling biomechanical criteria for hMSC and osteocyte survival, proliferation, differentiation, cell cycle, and retention. Methods: Primary hMSCs from four healthy donors were characterized by flow cytometry, immunofluorescence, and Western blotting. Cells were seeded onto Ti-6Al-4V scaffolds with graded porosity (P50–P90, 50–90% porosity) and cultured for 120 h. Viability and retention were measured by trypan blue exclusion, apoptosis and cell cycle by flow cytometry, and osteogenic differentiation by collagen I, osteocalcin, and Akt phosphorylation analyses. Collagen-embedded hMSCs and osteoblasts were used to assess migration and phenotype maintenance. Results: The densest scaffold, P50, consistently retained significantly more viable hMSCs and differentiated osteoblasts than P60 and P70 (p < 0.005 in both cases, One-way ANOVA analysis, significance level a = 0.05 followed by Bonferroni correction). Among the scaffold architectures investigated, P50 demonstrated the most favorable early cellular retention under the present experimental conditions. No scaffold-induced apoptosis or proliferation changes were detected. Osteogenic differentiation and Akt phosphorylation increased during culture. In collagen-containing scaffolds, cells migrated from the matrix and preferentially coated titanium struts, with P50 supporting superior attachment. Conclusions: Among the scaffold architectures investigated, the low-porosity/high-surface-area P50 design provided the most favorable early microenvironment for hMSC and osteoblast retention without cytotoxic effects. Full article

Background/Objectives: Gastric cancer (GC) remains a leading cause of cancer-related mortality, with metastasis being the primary driver of poor prognosis. The extracellular matrix glycoprotein tenascin-C (TNC) is implicated in tumor progression and metastasis, yet its regulatory mechanisms in GC remain poorly understood. Here, we identify RNA N6-methyladenosine (m6A) demethylase fat mass and obesity-associated protein (FTO) as a key modulator of TNC expression, promoting epithelial–mesenchymal transition (EMT) and GC metastasis. Methods: Integrating The Cancer Genome Atlas Stomach Adenocarcinoma (TCGA-STAD) and Gene Expression Omnibus (GEO) datasets (GSE246567, GSE181840, GSE163126, GSM4837878), we demonstrate that TNC is significantly upregulated in GC and correlates with worse clinical outcomes. Functional assays both in vitro and in vivo reveal that TNC knockdown suppresses GC cell proliferation, migration, invasion, and tumor metastasis, while overexpression of TNC rescues these effects. Mechanistically, FTO selectively demethylates TNC mRNA, reducing m6A modification at the 3’ untranslated region, thereby enhancing TNC mRNA stability and expression. RNA pull-down assays identify YTH domain family protein 2 (YTHDF2) as the m6A reader that recognizes the methylated site on TNC 3’UTR, and YTHDF2 knockdown partially rescues TNC expression and stability upon FTO loss. m6A RNA immunoprecipitation (MeRIP) and dual-luciferase reporter assays confirm FTO’s direct regulation of TNC via m6A demethylation. Furthermore, TNC promotes EMT by activating the phosphoinositide 3-kinase (PI3K)-AKT (protein kinase B) signaling pathway, as demonstrated by rescue experiments using the AKT activator SC79 and the PI3K inhibitor LY294002. Results: High FTO and TNC expression levels in clinical tissue samples correlate with poor patient survival. Conclusions: These findings reveal that the FTO/TNC/EMT axis represents a previously unrecognized epigenetic mechanism driving GC metastasis and suggest that targeting FTO-mediated m6A modification may provide a novel therapeutic strategy for GC patients with high metastatic potential. Full article

Background: In our companion study, the gingival mesenchymal stem cell (GMSC) secretome suppresses oxidative stress and induces apoptosis in primary oral squamous cell carcinoma (OSCC) cells, where those wet-lab results are themselves reported as preliminary; here we ask whether it also engages a proliferation × migration programme in patient tissue. The two arms differ in read-out type (apoptosis and redox there, transcript abundance here), not in opposed function. Methods: Primary OSCC cells received GMSC-conditioned medium (GMSC-CM) or indirect Transwell co-culture, assayed by RT-qPCR (VEGFA, TGFB1, MMP9, CXCL12, CCND1, PCNA, MYC, EGFR), MTT, and scratch-wound migration. Thirteen computational layers plus a GeoMx spatial verify-and-decide layer were applied to public data: TCGA-HNSC, GEO, CPTAC, single-cell inference, prognostic modelling, DepMap and LINCS L1000. Results: All eight transcripts were raised in all three arms (16 of 24 comparisons significant), indicating a pro-tumour transcriptional shift; metabolic activity was unchanged and wound closure was reduced under conditioned medium (p < 0.01). Of 1358 genes significant in all three modalities, 1219 (89.8%, 95% CI 88.0–91.3%) share direction against 25.0% expected by chance (3.59-fold; exact binomial p below machine precision). The pre-registered oral-cavity signature did not validate externally (GSE41613; C = 0.562), and no ligand–receptor pair survived permutation calibration. Conclusions: GMSC paracrine exposure induces a pro-tumour transcriptional programme in primary OSCC cells that replicates at patient level, without demonstrated functional or therapeutic consequence; gefitinib is nominated only as a hypothesis-generating candidate. The evidence rests on three primary cultures (n = 3), one GMSC donor preparation, one 24 h time point, a single reference gene, no cell-line authentication and no test of gefitinib; the study programme has concluded, and these experiments cannot be performed now. Full article

Background/Objectives: Colorectal cancer (CRC) remains a major cause of cancer-related mortality worldwide, largely owing to metastasis, recurrence, and resistance to conventional therapies. Natural-product-derived compounds represent an important source of anticancer drug candidates. This study investigated the anticancer activity of purified narciclasine, which was tentatively annotated as a major detected constituent of Lycoris sanguinea extract, and examined its association with HELLS-associated DNA damage responses in CRC cells. Methods: Human CRC cell lines HT29 and HCT116 were used to evaluate the cytotoxic and mechanistic effects of L. sanguinea extract and narciclasine. Cell viability, clonogenic growth, apoptosis, mitochondrial membrane potential, cell cycle distribution, migration, and invasion were assessed. Western blotting was performed to analyze apoptosis-, cell cycle-, epithelial–mesenchymal transition (EMT)-, and DNA damage-related proteins. UPLC-QTOF/MS analysis was used to identify major phytochemical constituents. HELLS knockdown and γ-H2AX immunofluorescence staining were conducted to examine the involvement of HELLS in narciclasine-associated DNA damage and apoptosis. Results: L. sanguinea extract reduced CRC cell viability, suppressed clonogenic growth, induced mitochondrial dysfunction and caspase-dependent apoptosis, and promoted G2/M cell cycle arrest with decreased cyclin B1 and increased p53/p21 expression. The extract also inhibited migration and invasion, accompanied by reduced β-catenin, N-cadherin, vimentin, slug, and snail expression and increased E-cadherin expression. UPLC-QTOF/MS tentatively annotated narciclasine as a major detected constituent. Purified narciclasine recapitulated several anticancer effects of the extract, accompanied by HELLS downregulation, increased γ-H2AX accumulation, reduced cyclin B1 expression, and enhanced PARP and caspase-3 cleavage. HELLS knockdown further sensitized CRC cells to narciclasine-associated γ-H2AX accumulation, G2/M arrest-associated signaling, and apoptosis. Conclusions: Narciclasine treatment was associated with HELLS downregulation, DNA damage-associated signaling, G2/M cell cycle arrest-associated changes, EMT suppression, and apoptosis in CRC cells. These findings suggest that narciclasine is a promising natural-product-derived lead compound for further preclinical evaluation in HELLS-associated CRC vulnerabilities. Full article

Cholangiocarcinoma (CCA) is an aggressive malignancy with limited treatment options and poor clinical outcomes. Kinesin light chain 1 (KLC1), a component of the kinesin-1 motor complex involved in intracellular transport, has been implicated in cancer biology; however, its role in CCA remains unclear. This study investigated the functional role and molecular alterations associated with KLC1 silencing in CCA. Analysis of publicly available datasets showed that KLC1 mRNA expression was significantly elevated in CCA tissues, and immunohistochemical images from the Human Protein Atlas demonstrated stronger KLC1 protein expression in tumor tissues. siRNA-mediated KLC1 knockdown markedly suppressed cell proliferation, migration, and invasion in KKU-213A and KKU-055 cells and altered the expression of epithelial–mesenchymal transition-associated proteins. Transcriptomic profiling identified 2074 differentially expressed genes following KLC1 knockdown. Functional enrichment analyses revealed significant alterations in cytoskeleton-associated processes and mitogen-activated protein kinase (MAPK) signaling. Protein–protein interaction network analysis identified interconnected gene networks associated with these pathways. Selected differentially expressed genes were validated by RT–qPCR, supporting the transcriptomic findings. Collectively, these results suggest that KLC1 contributes to aggressive phenotypes in CCA cells and is associated with transcriptomic alterations involving cytoskeletal regulation and MAPK signaling, highlighting KLC1 as a potential contributor to CCA progression and warranting further investigation. Full article

Non-small cell lung cancer (NSCLC) remains a leading cause of cancer-related mortality. Smoking is the primary etiological factor, but growing evidence suggests the involvement of estrogen in its development and progression, although its role remains unclear. This study explores: (i) the effects induced by estrogen, namely, 17β-estradiol (E2), alone or in combination with a mixture of inflammatory cytokines (Mix), in two human NSCLC cell lines, A549 and Calu1, and (ii) whether and how tumor cells can modulate the activation of normal lung fibroblasts. We found that E2 significantly enhances migration, invasion, and epithelial–mesenchymal transition in NSCLC cells, as well as their resistance to cisplatin, particularly in combination with Mix. Pharmacological inhibition of ERβ reversed the E2-induced effects, implicating ERβ in E2-mediated signaling. Furthermore, E2 increased autophagic flux and induced a shift toward secretory autophagy and the release of extracellular vesicles, which activated normal lung fibroblasts, as demonstrated by the increased expression of α-SMA, FAP, PDGFR-β, and PDPN. The clinical relevance of these data was supported by computational analyses revealing an elevated expression of the Mix gene signature, including TGF-β, IL-6, IL-8, CCXL-16, and ERβ, which was associated with shorter overall survival in NSCLC patients. This molecular profile was linked to the elevated expression of secretory autophagy genes and cancer-associated fibroblast markers. Validation in three large clinical cohorts (TCGA-LUNG, OAK and POPLAR) strengthens the clinical relevance of this E2-related pro-tumor axis while suggesting a promising therapeutic avenue for NSCLC patients. Full article

Background: Thin endometrium is a significant cause of infertility due to impaired regeneration, reduced receptivity, and insufficient angiogenesis. Mesenchymal stem cell-derived secretome (MSCsec) is a promising cell-free therapy because it contains bioactive molecules that promote tissue repair. This study evaluated the effects of Wharton’s jelly-derived MSC secretome (WJ-MSCsec) on human endometrial stromal cells (EnSCs) and endothelial cells in vitro. Methods: WJ-MSCs were isolated from umbilical cord tissue, characterized, and cultured under serum-free conditions. The concentrated secretome was analyzed using a human angiogenesis proteome array. EnSCs were isolated from endometrial biopsies. EnSC proliferation and migration in the presence of WJ-MSCsec were assessed using CCK-8 and scratch wound-healing assays. Pro-angiogenic activity of WJ-MSCsec was evaluated using a Matrigel tube formation assay with human umbilical vein endothelial cells (HUVECs). Results: Proteomic analysis of WJ-MSCsec revealed the presence of angiogenic, mitogenic, immunomodulatory, and chemotactic factors. Treatment with WJ-MSCsec significantly enhanced EnSC proliferation and accelerated wound closure. Furthermore, WJ-MSCsec markedly promoted endothelial tube formation, increasing total tube length, junction number, and mesh formation. Conclusions: WJ-MSCsec stimulates EnSC proliferation and migration while enhancing angiogenesis in vitro. WJ-MSCsec represents a promising cell-free therapeutic strategy for endometrial regeneration and reproductive disorders associated with impaired endometrial function. Full article

Background: Pyrotinib, an irreversible pan-HER tyrosine kinase inhibitor, has demonstrated substantial clinical efficacy in patients with HER2-positive breast cancer (BC). However, intrinsic and acquired resistance remain important challenges limiting therapeutic benefit, and reliable biomarkers for predicting pyrotinib response are currently unavailable. This study aimed to identify molecular determinants associated with pyrotinib resistance and uncover their underlying mechanisms. Methods: Pre-treatment tumour samples from an exploratory discovery cohort of 12 patients with HER2-positive BC receiving pyrotinib-containing neoadjuvant therapy were analysed by proteomic profiling. Differentially expressed proteins (DEPs) between the pathological complete response (pCR) and non-pCR groups were integrated with weighted gene co-expression network analysis and protein–protein interaction network analysis to identify candidate proteins. The prognostic relevance of the candidate genes was subsequently evaluated using 127 machine-learning strategies across three independent BC cohorts. Models were ranked according to the mean area under the receiver operating characteristic curve (AUC) across two evaluation cohorts, and SHapley Additive exPlanations (SHAP) analysis was performed separately in both cohorts to prioritise a candidate for subsequent investigation. In vitro and in vivo experiments were then conducted to evaluate the biological role of the prioritised candidate and its association with pyrotinib sensitivity. Finally, the association between pre-treatment SLC27A2 expression and pCR was evaluated in an independent, non-overlapping retrospective cohort of 103 patients receiving pyrotinib-containing neoadjuvant therapy. Results: Exploratory proteomic profiling of 12 pre-treatment tumour samples identified 617 DEPs between the pCR and non-pCR groups. Among 127 machine-learning strategies used to evaluate the prognostic relevance of the candidate genes, the glmBoost–random forest model achieved the highest mean AUC across the two evaluation cohorts (mean AUC = 0.678). SHAP analysis showed that SLC27A2 ranked second in GSE16446 and first in GSE48390 according to mean absolute SHAP values, supporting its prioritisation for subsequent investigation. Functional experiments showed that SLC27A2 promoted proliferation, migration, invasion, and epithelial–mesenchymal transition in HER2-positive BC cells. SLC27A2 knockdown enhanced pyrotinib sensitivity in vitro. In the xenograft experiment using female BALB/c nude mice, both SLC27A2 knockdown and pyrotinib treatment reduced tumour growth, and a significant interaction between the two factors was observed for endpoint tumour weight (p for interaction = 0.041). Mechanistically, SLC27A2 knockdown reduced lipid accumulation and PPARα expression, whereas pharmacological activation of PPARα partially attenuated the increase in pyrotinib sensitivity induced by SLC27A2 knockdown. Clinical validation further showed that high-pre-treatment SLC27A2 expression was independently associated with a lower likelihood of achieving pCR after pyrotinib-containing neoadjuvant therapy (OR = 0.10, 95% CI: 0.03–0.31, p < 0.001). Conclusions: SLC27A2 is a candidate factor associated with BC prognosis and reduced pyrotinib sensitivity in HER2-positive BC. Preclinical findings suggested that PPARα-related fatty acid metabolism may contribute to the association between SLC27A2 and pyrotinib response, while clinical validation showed that high-pre-treatment SLC27A2 expression was independently associated with a lower likelihood of achieving pCR following pyrotinib-containing neoadjuvant therapy. These findings support SLC27A2 as a candidate response-associated biomarker and potential therapeutic target, although further mechanistic investigation and external clinical validation are required. Full article

Background: As a member of the T-box transcription factor family, TBX18 was found to be involved in ESCC progression, while its role in regulating radiotherapy resistance in ESCC remains unclear. This study was designed to investigate the molecular mechanisms underlying the regulation of radioresistance in ESCC by TBX18. Methods: Sphere formation assay, Transwell invasion assay, and wound healing assay were conducted to show the influence of TBX18 on tumor stemness and epithelial–mesenchymal transition (EMT). Western blot, immunofluorescence, chromatin immunoprecipitation-qPCR (ChIP-qPCR) and dual-luciferase reporter assay were preformed to identify regulatory networks. A nude mouse xenograft tumor model was established to assess the regulatory effect of TBX18 and SIX1 on radioresistance of ESCC in vivo. Results: TBX18 expression was positively associated with stemness markers, including CD44, CD271, and SOX2. TBX18 promoted stemness-associated phenotypes, EMT, migration, invasion, and radioresistance in ESCC cells. Mechanistically, TBX18 directly bound to the SIX1 promoter and transcriptionally activated SIX1 expression. In turn, SIX1 enhanced TBX18 protein stability by suppressing ubiquitin–proteasome-mediated degradation, thereby forming a positive feedback loop. Functional rescue experiments demonstrated that the TBX18/SIX1 axis coordinately maintained stemness and EMT phenotypes and attenuated radiotherapy-induced apoptosis. In vivo studies further confirmed that TBX18 knockdown enhanced radiosensitivity, whereas SIX1 overexpression partially reversed this effect. In addition, immunohistochemical analysis revealed that TBX18 and SIX1 were significantly upregulated in ESCC tissues and positively correlated with each other. Full article

Regucalcin (RGN), also known as senescence marker protein-30, is a calcium-binding protein without an EF-hand motif that regulates intracellular calcium homeostasis and Ca²⁺-dependent enzyme activity. Its expression is reduced in tumor tissue relative to matched normal tissue across several cancer types, and restoring RGN in cancer cell lines suppresses proliferation, migration, and invasion through cell cycle arrest, reduced expression of matrix metalloproteinases and epithelial–mesenchymal transition regulators, and transcriptional reprogramming involving p53, p21, Rb, c-myc, and β-catenin. Attention has more recently turned to RGN outside the cell. RGN is detectable in serum and interstitial fluid, and recombinant RGN applied to cancer cells reproduces much of the suppression seen upon intracellular overexpression, which has prompted the proposal that extracellular RGN acts within the tumor microenvironment (TME). That proposal, however, rests on evidence that remains incomplete. RGN carries no classical signal peptide and its route of release is undefined; it has been reported in extracellular vesicle preparations but has not been shown to be a bona fide vesicular cargo; no receptor or proximal binding partner has been identified; and its effects on the non-malignant compartment of the TME—fibroblasts, immune cells, and endothelium—have not been tested directly. Here, we review the tumor-suppressive activity of RGN, distinguish demonstrated findings from inferred ones, and outline the experiments required to determine whether extracellular RGN constitutes a signaling axis in the TME or a pharmacological property of a recombinant protein. Full article

The kallikrein–kinin system (KKS) and the kallikrein-related peptidase (KLK) family are interconnected proteolytic networks that regulate inflammatory signaling, vascular permeability, extracellular matrix remodeling, and tissue barrier dynamics. Beyond their classical vasoactive and inflammatory functions, accumulating evidence indicates that kinin peptides, including bradykinin, Lys-bradykinin, and their des-Arg⁹ metabolites, together with selected KLKs, modulate cell–cell and cell–extracellular matrix adhesion. Through B1 and B2 kinin receptor activation, the KKS influences endothelial adhesion molecule expression, leukocyte integrin activation, neutrophil trafficking, focal adhesion kinase/Src signaling, cytoskeletal remodeling, and matrix metalloproteinase activity. In parallel, KLKs directly reshape the adhesive microenvironment by cleaving junctional proteins, including E-cadherin and desmosomal components, and extracellular matrix substrates such as fibronectin, laminin, vitronectin, fibrinogen, and collagens. These coordinated actions affect epithelial and endothelial barrier integrity, leukocyte transmigration, angiogenesis, fibrosis, epithelial–mesenchymal transition, tumor cell migration, invasion, and metastatic dissemination. This review critically summarizes current evidence linking KKS and KLK activity to adhesion-dependent processes in inflammation and cancer, emphasizing how proteolytic signaling may either preserve tissue homeostasis or promote pathological barrier disruption depending on cellular context, receptor expression, protease activity, and microenvironmental cues. Understanding these mechanisms may refine the identification of adhesion-related biomarkers and support the development of targeted therapeutic strategies for inflammatory disorders, fibrotic remodeling, and cancer progression. Full article

Metastasis-associated protein 1 (MTA1), an oncogenic transcriptional regulator, is overexpressed in canine urothelial carcinoma (UC) and is associated with aggressive clinicopathological features. However, its functional role and molecular mechanisms in canine UC remain poorly understood. Here, we investigated the contribution of MTA1 to epithelial-to-mesenchymal transition (EMT) and its interaction with BRAF signaling. MTA1 silencing in two canine UC cell lines significantly inhibited cell proliferation, cell survival, migration, and xenograft tumor growth. Mechanistically, MTA1 knockdown reduced the expression of MTA2, MTA3, and COX2, while producing unexpected changes in key EMT regulators, including Snail, Slug, and Cyclin D1, suggesting the activation of compensatory signaling pathways. MTA1 silencing also decreased mutant BRAF expression in AxA cells while increasing wild-type BRAF expression in SH cells, indicating context-dependent regulation of BRAF signaling. In AxA cells, reduced AKT phosphorylation following MTA1 knockdown further supports functional crosstalk between the BRAF and MTA1/AKT signaling pathways. Collectively, these findings identify MTA1 as a critical regulator of canine UC progression and reveal complex signaling interactions that support its potential as a therapeutic target for canine UC. Full article

Background/Objectives: Glioblastoma is an aggressive primary brain tumor that develops within a chronically low-oxygen microenvironment, yet most preclinical studies are performed under atmospheric oxygen conditions that poorly reflect in vivo physiology. This study investigated how sustained culture under physiological oxygen tension (physioxia) influences glioblastoma cell behavior, signaling, and therapeutic response. Methods: Multiple patient-derived glioblastoma models were cultured under normoxia (21% O₂) or sustained physioxia (5% O₂) for at least seven days before experimentation. Cell migration, proliferation, cell cycle distribution, expression of the epithelial-to-mesenchymal transition-associated transcription factor Slug (SNAI2), PDGFRβ-associated signaling, and sensitivity to 5-fluorouracil were evaluated using transwell migration assays, cell counting, flow cytometry, RT-qPCR, immunoblotting, and BrdU incorporation assays. To examine the effects of oxygen history, we established and maintained additional patient-derived cultures grown under physioxia. Results: Sustained physioxia consistently increased migration across all glioblastoma models while reducing proliferation in normoxia-adapted cell lines through increased G0/G1 cell cycle arrest. Physioxia significantly increased Slug expression in all models and enhanced PDGFRβ, AKT, and ERK phosphorylation in a cell line-dependent manner. Therapeutic sensitivity to 5-fluorouracil was altered, with physioxia conferring increased resistance in some glioblastoma models but not universally. Patient-derived cell lines cultured continuously under physioxia retained enhanced migratory capacity and exhibited increased proliferation compared to cells grown in normoxia, suggesting that prior oxygen exposure influences proliferative responses while the pro-migratory phenotype remains conserved. Conclusions: Physiological oxygen tension is a major regulator of glioblastoma cell behavior, influencing migration, proliferation, signaling, and therapeutic response. These findings demonstrate that conventional normoxic culture conditions can obscure biologically relevant phenotypes and support the consideration of physioxia in experimental design. Such technical changes may improve the physiological and translational relevance of preclinical glioblastoma research. Full article

Background/Objectives: Adenomyosis is a chronic gynecological disorder characterized by the presence of endometrial tissue within the myometrium, causing pelvic pain, abnormal uterine bleeding, and infertility. Despite its high prevalence, the molecular mechanisms underlying disease initiation and progression remain incompletely understood. Current evidence implicates disruption of the endometrial–myometrial interface, epithelial–mesenchymal transition, and progesterone resistance in driving tissue invasion and remodeling. Diagnosis relies mainly on imaging modalities, while reliable non-invasive biomarkers are lacking. MicroRNAs, as stable post-transcriptional regulators of gene expression, have emerged as key modulators of proliferation, inflammation, and hormonal signaling, and represent promising candidates for novel diagnostic strategies. Methods: A systematic review was conducted in accordance with PRISMA guidelines and registered with PROSPERO (CRD42025637752). A comprehensive search of the Medline, Embase, Scopus, and Cochrane databases was performed in April 2025. Studies investigating miRNA expression in patients with adenomyosis compared with controls were included. The quality of the studies and the risk of bias were assessed using the Newcastle–Ottawa scale. Two reviewers independently performed study selection, data extraction, and quality assessment. Results: Twenty-seven studies published between 2015 and 2025 met the inclusion criteria. Thirty-nine distinct miRNAs were reported as significantly dysregulated in adenomyosis. Recurrently altered miRNAs included let-7a, miR-145, miR-10b, miR-30c-5p, miR-141-3p, miR-143, and miR-191. Functional analyses have consistently implicated miRNAs in key pathogenic pathways, including Hippo-YAP, PI3K/AKT, MAPK/ERK, JAK/STAT, and Wnt/β-catenin signaling. These alterations were associated with enhanced epithelial–mesenchymal transition, increased cellular proliferation and migration, progesterone resistance, chronic inflammation, and immune modulation. Emerging evidence highlights exosomal and circulating miRNAs as promising non-invasive biomarkers, with a few studies already demonstrating diagnostic potential using serum, plasma, or urine samples. However, substantial heterogeneity in tissue types, sampling timing, and analytical methods precluded meta-analysis. Conclusions: MiRNAs play a central role in the molecular pathogenesis of adenomyosis and show strong potential as non-invasive diagnostic biomarkers. However, large-scale validation studies and standardized methodologies are required before clinical implementation. Full article