Methylated septin9 (septin9-mC) is a well-validated biomarker for colorectal cancer screening, and accurate detection of such site-specific methylated DNA holds significant clinical value for early disease diagnosis. However, conventional methods suffer from cumbersome pretreatment, DNA degradation risks, and poor performance in low-abundance samples. Herein, we report a synergistic iontronic sensing platform integrating methylation-sensitive restriction enzyme (AciI), CRISPR/Cas12a, Ag-DNAzyme, and Au/Pt heterometallic nanozyme for highly sensitive and specific detection of septin9-mC. AciI selectively cleaves unmethylated septin9 (septin9-C) while sparing septin9-mC, and intact septin9-mC activates Cas12a trans-cleavage activity to trigger catalytic hairpin assembly (CHA), generating Ag-DNAzyme. Activated Ag-DNAzyme induces detachment of Au/Pt nanoparticles from anodic aluminum oxide membranes, reducing the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to positively charged oxTMB and altering ion transport fluxes in nanochannels, which is read out via current-voltage characteristics. The linear range is 100 aM to 10 nM with a detection limit of 34.0 aM. This method effectively distinguishing colorectal cancer cells from human colonic epithelial cells and colorectal cancer patients from healthy individuals, showing excellent performance in real sample analysis. The proposed method provides a dependable tool for site-specific methylation detection with promising applications in biological research and clinical diagnosis.

Cell adhesion tightly controls cell proliferation and homeostasis in stratified epithelia by mechanisms that remain unclear. Focal adhesion kinase (FAK) transduces cell adhesion signals, is frequently deregulated in epithelial cancer, and it has been associated with proliferation and resistance to treatments. The mechanisms by which FAK controls the epithelial cell cycle are still intriguing. We previously unraveled a mitosis-differentiation checkpoint that is the limiting factor in the keratinocyte cell cycle. To investigate whether FAK plays a role in this checkpoint, we inactivated the protein in normal human oral keratinocytes by specific shRNAs or by the specific inhibitor defactinib. Inactivation of FAK very rapidly and strikingly blocked entry into mitosis and triggered a differentiation response. This response was independent of DNA damage. Tumor suppressor P53 was induced shortly after inhibition of FAK, while mitotic Cyclin B was not translocated into the nucleus. Human epidermal N-TERT cells that were synchronized in prometaphase failed to execute mitosis. Concomitant inhibition of FAK-downstream Rho-associated kinase (Rock) rescued mitotic progression. The results unveil a rapid Rock-dependent mitosis switch upon inactivation of FAK, inducing terminal differentiation, pointing at a mitotic automatic mechanism of epithelia to suppress suprabasal proliferation of precancerous cells.

Structural insights into frog skin-derived cyclic peptides as selective matriptase inhibitors.

The study aimed to investigate polycomb group factor 1 (PCGF1) expression in colon cancer and its mechanism of action in the proliferation and migration of colon cancer cells. In this study, PCGF1 expression in colon cancer and the association between PCGF1 expression and the survival prognosis of patients with colon cancer were analyzed using the cancer genome atlas (TCGA) database. Furthermore, PCGF1 expression in normal colon epithelial cells and colon cancer cells was examined. A PCGF1 knockdown cell model was developed using HCT116 and HT29 cell lines. The effect of PCGF1 on cell proliferation was evaluated via CCK-8 and clone formation assays. In addition, the effect of PCGF1 on cell invasive and migratory abilities was explored via scratch and Transwell experiments, and the expression levels of EMT-related proteins was analyzed via immunoblotting assay. PCGF1 expression significantly increased in colon cancer tissues. This expression was also found to be closely associated with tumor staging of patients with colon cancer. High PCGF1 expression showed negative correlation with overall survival rate in these patients. The proliferative ability of HCT116 and HT29 cells knocked down by PCGF1 weakened, colony formation was reduced, and cell invasive and migratory capabilities diminished. Meanwhile, the expressions of EMT-related proteins, such as N-cadherin and Snail1, were remarkably downregulated in PCGF1 knockdown cells, whereas that of E-cadherin was significantly upregulated. Moreover, PCGF1 knockdown led to a strongly decrease in the phosphorylation levels of PI3K, Akt and mTOR proteins and in the levels of β-catenin and c-Myc. PCGF1 may promote the proliferation and EMT process in HCT116 and HT29 cells by activating the Wnt/β-catenin and PI3K/Akt/mTOR signaling pathway.

Effects along the epithelial-mesenchymal biointerface in direct cell self-organisation: Multiscale theoretical analysis.

Interaction of tumor cells and Cancer-associated fibroblasts in radiation-induced senescence.

Background: The five-year survival rate of patients with ovarian cancer remains less than 50%, secondary to chemotherapy resistance. Purpose: This study aims to evaluate the effects of itraconazole as a supplementary treatment with paclitaxel and carboplatin on malignancy response and in preventing the initial development of chemoresistance in chemotherapy-naïve patients with advanced ovarian epithelial cancer. Method: This randomized placebo-controlled double-blind study involved 60 chemotherapy-naïve patients with advanced epithelial ovarian malignancy who were randomized into two arms; the placebo and itraconazole groups. The placebo group received six chemotherapy cycles and four inactive capsules, while the itraconazole group received six chemotherapy cycles and 400 mg oral itraconazole for five days per cycle. Results: Following completion of six chemotherapy cycles and when contrasted with the control arm, the itraconazole arm demonstrated statistically significant improvements in tumor response. The objective response rate was 80% in the itraconazole group compared with 47% in the placebo group (p = 0.015), while the disease control rate was 100% versus 80%, respectively (p = 0.023). The median progression-free survival (PFS), defined as the time point at which 50% of patients experienced disease progression or death, was 13.5 months for the overall study population. PFS was evaluated as a fixed-time endpoint at 18 months following completion of chemotherapy for the overall study population. Progression-free survival was significantly improved in the itraconazole group, with 70% of patients remaining progression-free compared with 26.7% in the placebo group (p = 0.001). Also, the itraconazole group produced significant declines in the serum levels of CA-125 (p = 0.005) and p-glycoprotein (p = 0.042) with significant elevation in VEGFR-2 (p = 0.006) as compared to the control group. Itraconazole was safe and its use was associated with a significant improvement in the quality of life (QOL). Conclusions: Itraconazole could represent a promising add-on therapy to enhance tumor response to chemotherapy in patients with ovarian cancer.

Piezo1 ion channels play a crucial role in apoptosis regulation in human breast cancer cells (MCF-7), and this study evaluates the effects of Piezo1 agonist (Yoda1), inhibitor (GsMTx4), and ultrasound microbubble (USMB) treatment on cellular apoptosis pathways. In this research, in vitro cultures of normal breast epithelial cells (MCF-10A) and cancer cell lines (MCF-7, MDA-MB-231) were analyzed by Western blotting to determine Piezo1 protein levels, with MCF-7 selected for further analysis. Groups included control (untreated), Yoda1, USMB, GsMTx4, and USMB+GsMTx4, and apoptosis rates were measured via flow cytometry. Levels of apoptosis-related proteins (Bcl-2, Bax), endoplasmic reticulum stress proteins (GRP-78, Caspase 12), and mitochondrial pathway proteins (Cyt-c, Caspase 3, Caspase 9) were quantified, while JC-1 and Ca2+ fluorescent probes were used to assess mitochondrial membrane potential and intracellular Ca2+ concentration. Results showed MCF-7 cells expressed the highest Piezo1 levels. Yoda1 and USMB both markedly increased apoptosis, enhanced ER stress, and induced the mitochondrial apoptosis pathway in comparison to control, while GsMTx4 had the opposite effect and USMB reversed GsMTx4's phenotype. The USMB group exhibited the lowest mitochondrial membrane potential and the highest Ca2+ fluorescence intensity. These findings indicate that USMB activates ER stress via Piezo1, induces mitochondrial dysfunction, elevates intracellular Ca2+, and thereby promotes apoptosis in breast cancer cells.​​.

There remains a need for animal models with human translatability in lung cancer (LC) research. Findings in pigs can have a substantial impact owing to their similar anatomy and physiology to humans. Here we present a bronchoscopically induced LC model in Oncopigs carrying inducible KRASG12D and TP53R167H mutations. A total of 12 Oncopigs underwent 29 injections via flexible bronchoscopy: 18 adenovirus-Cre recombinase gene (AdCre) inductions were performed endobronchially (n = 6) and transbronchially (n = 12), and 11 control injections were performed without AdCre. Oncopigs underwent serial chest CT with clinical follow-up for 29 weeks. Lung and organ tissues underwent histopathology, immunohistochemistry and RNA sequencing with comparative analysis to human LC data. All 18 sites of AdCre injections had lung consolidations on computed tomography imaging. Inductions led to an overall success rate of 77.8%, including both invasive cancer (61.1%) and carcinoma in situ (16.7%). Transbronchial injections led to histopathologic invasive cancer and/or carcinoma in situ in 11/12 (91.7%) and invasive cancer in 8/12 (66.6%). Endobronchial inductions led to invasive cancer in 3/6 (50%). A soft tissue metastasis was observed in one Oncopig. Immunohistochemistry confirmed the expression of Pan-Cytokeratin (Pan-CK)+ in epithelial cancer cells, with macrophage and T cell infiltration in the tumor microenvironment. Transcriptome comparison showed 54.3% overlap with human LC, while KRAS-mutant mouse LC had 29.88% overlap with human LC. The immunocompetent Oncopig model has a high rate of LC following bronchoscopic transbronchial induction. An overlap of the Oncopig LC transcriptome with the human LC transcriptome was noted. This pig model of LC is expected to have high clinical translatability.

How do vibration stimulation frequencies affect the nonlinear dynamics and mechanical characterization of breast cancer cells?

Lipid nanoparticles (LNPs) are useful carriers for therapeutic siRNA delivery, yet their clinical efficacy remains constrained by insufficient cellular uptake. Here, using a genome-wide CRISPR knockout screen, multiple genetic modulators of LNP uptake is uncovered, with PIK3CA emerging as a top druggable target. Pharmacologic inhibition of PIK3CA with BAY1082439 - a clinically evaluated small molecule - significantly enhances LNP uptake, siRNA delivery, and gene silencing across diverse epithelial cancer cell lines in vitro. Co-administration of BAY1082439 with siRNA-loaded LNPs also better suppressed tumor growth and reduced liver inflammation in vivo, respectively. These findings establish PIK3CA inhibition as a broadly applicable strategy to boost LNP-mediated RNA interference and highlight the promise of combining functional genomics with nanomaterials to advance RNA-based therapeutics.

The therapeutic limitations of conventional anticancer agents, particularly in treating mucosal and epithelial malignancies such as breast and colorectal cancers, necessitate the development of advanced drug delivery systems. This study explores the integration of benzimidazole-1,2,3-triazole hybrids into chitosan/polyvinyl pyrrolidone nanogels to enhance pharmacological performance and target specificity. Comprehensive physicochemical characterization confirmed successful encapsulation and favorable nanogel properties, including controlled particle size, stability, and morphology. Among the tested formulations, the 12ng nanogel demonstrated superior cytotoxicity against the epithelial cancer cells, MDA-MB-231 (IC50 = 3.13 ± 0.22) and Caco-2 (IC50 = 3.64 ± 0.25) µM, surpassing the reference drug staurosporin. Enzymatic assays revealed potent inhibition of topoisomerase I and II, as well as tubulin polymerization. Flow cytometry and apoptosis analysis confirmed G0/G1 cell cycle arrest and late-stage apoptosis induction. Molecular docking studies supported the strong binding affinity (−9.50 kcal mol−1) of 12ng to topoisomerase I, validating its' multitarget mechanism. These findings underscore the potential of nanogel-delivered benzimidazole-1,2,3-triazole hybrids as promising candidates for targeted cancer therapy.

Extracellular vesicle and particle-based blood test for ovarian cancer screening of average risk population: a promising nested case control study using preclinical samples.

65 How do patients with severe oral epithelial cancer perceive chemoprevention and a chemoprevention clinical trial?: Embedded qualitative study within the SAVER trial

Quantitative analysis of p21 expression and its prognostic associations in breast cancer epithelium

Nasopharyngeal carcinoma (NPC), a type of epithelial cancer, is frequently found in Southeast Asia and Southern China. The Epstein-Barr Virus (EBV) significantly contributes to the development of NPC by producing latent proteins, particularly Latent Membrane Protein 1 (LMP-1). This protein acts as an oncoprotein, activating several signaling pathways, including NF-κB, JAK/STAT, and PI3K/Akt. Consequently, this factor facilitates cellular proliferation, angiogenesis, and the inhibition of apoptosis. A multitude of investigations have established a correlation between the expression of LMP-1 and both the malignancy of tumors and the clinical outcomes of individuals afflicted with KNF. LMP-1 detection can be achieved through immunohistochemistry (IHC), polymerase chain reaction (PCR), or in situ hybridization (ISH) techniques, each exhibiting varying degrees of sensitivity and specificity. Furthermore, LMP-1 serves as a critical diagnostic and prognostic indicator, and it also presents a potential target for molecular interventions in nasopharyngeal cancer.

BackgroundRecent studies have found the presence of post- hysterectomy atypical glandular cells in upper vagina in ~ 30% patients of Endometrial Cancer (EC) with implications of causing isolated vaginal recurrence. The primary objective was to determine the effectiveness of cervical canal and fallopian tube occlusion in preventing intra-operative tumour spillage in EC.MethodsA Prospective Interventional Single Institutional pilot study with random allocation to two arms using computer- generated random number sequence was conducted between January 2022 to Feb 2023. Patients with histologically confirmed epithelial endometrial cancer, including endometrioid carcinoma, papillary serous, clear-cell, squamous, mucinous, undifferentiated, and carcino-sarcoma with surgery as primary definitive treatment modality were included. Spillage was detected using vaginal smear and peritoneal wash cytology taken pre- and post- hysterectomy. In interventional arm, cervical os and fallopian tube were occluded using silk sutures as additional steps during surgery. Rate of negative to positive conversion of vaginal smear and peritoneal wash cytology obtained before and after hysterectomy in the interventional arm was compared with control arm who underwent surgery with standard surgical steps. Results: Out of 33 eligibile patients, 3 were excluded due to double primaries and history of radiation pelvis. The two arms of 15 patients each were comparable with respect to clinico-pathological characteristics. The most common histology, grade, degree of myometrial invasion, LVSI, stage, and mean tumour diameter was endometrioid variety (100%), low-grade (83.3%), & >1/2 (50%), negative (83.3%), 1 A (41.7%), and 4 cm in interventional arm and endometrioid (100%), low grade (76.9%), >1/2 (84.6%), negative (92.3%), 1B (84.6%), and 4.5 cm in non- interventional arm respectively. None of the patient in either arm had negative to positive conversion of vaginal and/or peritoneal cytology. Conclusion: We observed no difference of cervical and tubal occlusion on tumour cell spillage in EC.Trial registrationCTRI/2022/05/042348.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12957-025-04121-5.

This study aims to investigate the heterogeneity of cancer-associated fibroblasts (CAFs) and smooth muscle cells (SMCs) and their interactions with malignant epithelium in gastric cancer (GC), with the goal of identifying potential therapeutic targets. We performed a multi-omics analysis integrating single-cell RNA sequencing and spatial transcriptomics, complemented by trajectory inference, regulon activity mapping, ligand-receptor interaction modeling, and survival association in external cohorts. Two malignant epithelial programs emerged-Tumor Cell 1 (TC 1) (wound-healing/proliferative) and Tumor Cell 1 (TC 2) (highly cycling/metabolic). In TCGA-STAD (The Cancer Genome Atlas - Stomach Adenocarcinoma), high TC 1 scores associated with worse overall survival (P < 0.01), whereas TC 2 showed a nonsignificant trend. The stroma comprised nine populations; tumors were enriched for RGS5⁺ SMCs and FAP⁺ fibroblasts, with depletion of homeostatic PI16⁺ fibroblasts. Pseudotime traced a PI16⁺→CCL11⁺→PDGFRA⁺→FAP⁺ continuum featuring late NF-κB/STAT activation and extracellular matrix (ECM)-remodeling; SMCs rewired from contractile MYH11⁺ to angiocrine RGS5⁺ states. Ligand-receptor analysis indicated asymmetric tumor-stroma crosstalk: TC 1 preferentially engaged vaso-regulatory/EGFR-immune signaling to activate CAFs and SMCs, while TC 2 amplified PDGF/MDK growth-factor circuits; stromal feedback via WNT/NRG/HGF/TGF-β reinforced malignant programs. Spatial maps confirmed EPCAM-high tumor nests encased by COL1A2/FAP⁺ fibroblastic shells interwoven with ACTA2/RGS5⁺ strands. This study highlights the critical role of CAFs and SMCs heterogeneity and their interactions within the GC TME (Tumor microenvironment). Targeting stromal pathways such as PDGF, TGF-β, and NF-κB signaling, immunomodulating CAFs, and disrupting SMC-derived vascular niches may improve therapeutic responses. Further experimental validation of ligand-receptor pairs and spatial determinants is warranted.

BackgroundMalignant ascites (MA) is one of the major complications in advanced epithelial cancer patients and is associated with poor prognosis, poor quality of life, and severe symptoms. No efficient medicine is available for treating MA worldwide. Only paracentesis is recommended by the guidelines in most countries, but with limited efficacy and a short control time. Thus, novel treatments are needed to control MA.MethodsAn anti-EpCAM × anti-CD3 bispecific antibody, M701, was constructed as a T-cell engager to eliminate tumor cells in the peritoneal cavity. A phase II study was performed to evaluate the efficacy and safety of the intraperitoneal (IP) infusion of M701 in advanced epithelial tumor patients with moderate-to-large-scale MA. In this study, 84 patients were enrolled, with 43 in the M701 group receiving paracentesis and IP M701 infusion and 41 in the control group receiving paracentesis alone.ResultsThe primary endpoint, median puncture-free survival (PuFS), was 75 days in the M701 group and 25 days in the control group, with a significant difference (p = 0.0065). Subgroup analysis indicated that different types of cancer, including gastric, colorectal, and ovarian cancers, all benefited from the M701 infusion. Patients with higher relative lymphocyte counts (≥ 13%) at baseline received better effects. Compared to patients in the control group, the overall survival (OS) of patients in the M701 group was certain extended (mOS 110 days vs. 76 days, p = 0.1443, HR = 0.68). The 6-month survival rates were 33.3% and 12.1% in the two groups, respectively. No additional serious adverse events (SAEs) were detected in the M701 group. The most frequent treatment-related adverse events were anemia and low white blood cell count, which were manageable. M701 infusions did not cause a greater risk than paracentesis alone in the control arm, while all patients were administered systemic treatment.ConclusionWhen treated with M701, patients with MA had significantly longer puncture intervals and a trend of extended survival time. The results were encouraging for patients with MA. A phase III clinical trial of M701 aimed at further validation is ongoing.Supplementary InformationThe online version contains supplementary material available at 10.1186/s40164-025-00727-3.

Cholangiocarcinoma is a very deadly epithelial cell cancer with poor clinical outcome. Cellular senescence plays a vital role in the oncogenesis and the aggressiveness of cholangiocarcinoma. Integrative machine learning procedure including 10 methods (random survival forest, elastic network, Lasso, Ridge, stepwise Cox, CoxBoost, partial least squares regression for Cox, supervised principal components, generalized boosted regression modelling, and survival support vector machine) was performed to construct a cellular senescence-related signature (CSS) for cholangiocarcinoma. Cellular experiment was used to verify the biological function of hub gene. The optimal prognostic CSS developed by Lasso method served as an independent risk factor and had a stable and powerful performance in predicting the overall survival rate in cholangiocarcinoma, with the AUC of 1-, 3-, and 5-year ROC curve being 0.957, 0.929 and 0.928 in TCGA cohort. Low CSS score indicated with a lower tumor immune dysfunction and exclusion score, lower tumor microsatellite instability score, lower immune escape score, lower MATH score, and higher tumor mutation burden score in cholangiocarcinoma. Down-regulation of EZH2 inhibited the proliferation, colony and promoted apoptosis of cholangiocarcinoma cell. Integrative machine learning analysis was performed to construct a novel CSS in cholangiocarcinoma. This CSS acted as an indicator for predicting the prognosis and immunotherapy benefits of cholangiocarcinoma patients.