Journal: Cell Reports Medicine

Article Title: Early-stage multi-cancer detection through a plasma extracellular vesicle protein signature

doi: 10.1016/j.xcrm.2026.102694

Figure Lengend Snippet: Graphical summary of the study In the Discovery phase, a 4-protein biomarker panel (THBS1, NID1, PTX3, and VCAN) was discovered by proteomic analysis of sEVs derived from an isogenic HBEC model using mass spectrometry. The biomarker panel was tested in sEVs isolated from 22 cancer cell lines by ELISA. In the Validation phase, a cohort consisting of 250 healthy individuals and 514 patients with multiple cancers was recruited to assess the performance of the biomarker panel. Plasma/serum sEVs were isolated and analyzed by ELISA. In the Translation phase, a multiplex microfluidic device incorporating SERS was developed for simultaneous profiling of the 4-protein biomarker panel in a lung cancer screening setting. The expression levels of 4 proteins are reflected by the Raman intensities of the corresponding Raman reporters.

Article Snippet: Human VCAN ELISA Kit , Novus Biologicals , Cat# NBP2-75353.

Techniques: Biomarker Discovery, Derivative Assay, Mass Spectrometry, Isolation, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Multiplex Assay, Expressing

Journal: Cell Reports Medicine

Article Title: Early-stage multi-cancer detection through a plasma extracellular vesicle protein signature

doi: 10.1016/j.xcrm.2026.102694

Figure Lengend Snippet: Transformation-induced changes to the protein composition of cell-derived sEVs (A) The morphology of isolated sEVs was assessed using transmission electron microscopy. Images of normal and transformed HBEC-derived sEVs (scale bars, 200 nm). (B) Nanoparticle analysis using tunable resistive pulse sensing of sEVs isolated from HBECs demonstrates that the majority of sEVs have a size range between 30 and 150 nm, and that transformation does not result in an increase in sEV secretion. (C) Western blot of sEVs from HBECs demonstrating the presence of sEV proteins HSP70 and CD63 and the absence of the cell marker calnexin. (D) Label-free mass spectrometry identified 148 proteins with greater abundance in sEVs derived from transformed HBECs (FDR <0.02), of which 15 were annotated as extracellular proteins. (E) Mass spectrometry results were confirmed using ELISA for THBS1, NID1, PTX3, and VCAN in sEVs derived from normal and transformed HBECs. (F) sEVs derived from 22 cancer cell lines including NSCLC (SKMES1, H1650, HCC4006, and H2170), glioblastoma ([GBM], D54, D270, U87, and U118), colorectal cancer ([CRC], HT29 and SW620), breast cancer ([BCa], BT549, MDA231, and MDA436), prostate cancer ([PCa], PC3 and LNCaP), melanoma ([MEL], A375, MAMEL65, and SKMEL28), esophageal cancer ([ECa], OE19), and ovarian cancer ([OVA], A2780, CAOV3, IGROV1, and OVCAR8) showed a clear increase in expression of THBS1, NID1, PTX3, and VCAN in relation to the average levels of sEVs from normal cells ([HBEC] 30KT, HOSE 6.3, and HOSE 17.1). Samples in mass spectrometry and ELISA were measured in triplicate. See also and .

Article Snippet: Human VCAN ELISA Kit , Novus Biologicals , Cat# NBP2-75353.

Techniques: Transformation Assay, Derivative Assay, Isolation, Transmission Assay, Electron Microscopy, Tunable Resistive Pulse Sensing, Western Blot, Marker, Mass Spectrometry, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Cell Reports Medicine

Article Title: Early-stage multi-cancer detection through a plasma extracellular vesicle protein signature

doi: 10.1016/j.xcrm.2026.102694

Figure Lengend Snippet: The transformed sEV signature accurately diagnoses cancer in patient plasma (A) The expression levels of THBS1, NID1, PTX3, and VCAN in plasma derived from cancer patients are increased in comparison to healthy controls. Samples were measured in triplicate. Lines in dot plots represent median values. (B) ROC curves of classification of each cancer type compared to healthy controls demonstrate excellent diagnostic capability of the 4-protein sEV biomarker panel with an AUC of 0.91–1. (C) The sensitivity of the diagnostic sEV signature for each cancer type was evaluated at a fixed specificity of 90%, 95%, and 99%. Error bars represent 95% confidence intervals. Healthy ( n = 250), NSCLC ( n = 139), glioblastoma ([GBM], n = 57), colorectal cancer ([CRC], n = 42), prostate cancer ([PCa], n = 30), melanoma ([MEL], n = 100), gastric cancer ([GCa], n = 19), esophageal cancer ([ECa], n = 98), small cell lung cancer ([SCLC], n = 29). See also and .

Article Snippet: Human VCAN ELISA Kit , Novus Biologicals , Cat# NBP2-75353.

Techniques: Transformation Assay, Clinical Proteomics, Expressing, Derivative Assay, Comparison, Diagnostic Assay, Biomarker Discovery

Journal: Cell Reports Medicine

Article Title: Early-stage multi-cancer detection through a plasma extracellular vesicle protein signature

doi: 10.1016/j.xcrm.2026.102694

Figure Lengend Snippet: Evaluation of a multiplex microfluidic device applicable for liquid biopsy testing in a cancer screening setting (A) Clinical follow-up by CT imaging of 2 representative benign patients B1 and B2. Red arrows indicated nodules in patients’ lungs. B1 had a granuloma-cryptococcal infection, and the lesion was found less dense after 26 months. B2 had a lesion in the vicinity of emphysema, which resolved after 21 months. (B) Schematic of multiplex microfluidic device consisting of a pair of asymmetric circular electrodes. Electrodes are conjugated with an anti-THBS1 antibody to capture cancer-derived sEVs. SERS nanotags carrying designated Raman reporters and paired target antibodies (against THBS1, NID1, PTX3, and VCAN) are used for labeling captured sEVs and then analyzed by SERS mapping. (C) Representative false-color SERS spectral images demonstrating an enrichment of THBS1, NID1, PTX3, and VCAN in early-stage NSCLC patients (M1 and M2) compared to patients with benign diseases (B1 and B2). Scale bars, 10 μm. (D) The Raman intensity of each biomarker THBS1, NID1, PTX3, and VCAN in benign ( n = 27) and early-stage NSCLC ( n = 41) patients. a.u., arbitrary units. Samples were measured in triplicate. Lines in dot plots represent median values. (E) ROC curve of logistic regression classification indicating an AUC of 0.85 in detecting early-stage NSCLC cases compared to benign cases in a cancer screening setting. (F) The confusion matrix of the multiplex microfluidic device. See also and and .

Article Snippet: Human VCAN ELISA Kit , Novus Biologicals , Cat# NBP2-75353.

Techniques: Multiplex Assay, Imaging, Infection, Derivative Assay, Labeling, Biomarker Discovery

Journal: Cell Reports Medicine

Article Title: Early-stage multi-cancer detection through a plasma extracellular vesicle protein signature

doi: 10.1016/j.xcrm.2026.102694

Figure Lengend Snippet: Evaluation of the multiplex microfluidic device in a longitudinally monitored cohort of pre- and post-surgery NSCLC patients (A) Representative false-color SERS spectral images demonstrating a decrease of THBS1, NID1, PTX3, and VCAN in post-surgery NSCLC patients (P2 post and P12 post) compared to paired pre-surgery (P2 pre and P12 pre) patients. Scale bars, 10 μm. (B) Heatmap showing the log 2 fold changes in Raman intensities of THBS1, NID1, PTX3, and VCAN in post-surgery NSCLC patients ( n = 12), relative to their paired pre-surgery samples. Samples were measured in triplicate. Negative values (blue) indicate decreased expression after surgery, while positive values (red) indicate increased expression. See also .

Article Snippet: Human VCAN ELISA Kit , Novus Biologicals , Cat# NBP2-75353.

Techniques: Multiplex Assay, Expressing

Journal: Acta Histochemica et Cytochemica

Article Title: Role of MeCP2 in Shaping the Histopathological Heterogeneity of Ampullary Carcinoma

doi: 10.1267/ahc.25-00049

Figure Lengend Snippet: Visium-based transcriptomic clustering and spatial distribution of MeCP2-associated genes. ( a ) Visium spatial transcriptomic analysis identified five transcriptionally defined spatial clusters using unsupervised principal component analysis followed by graph-based Louvain clustering (Seurat v5.1.0). After clustering, the biological identities of the clusters were interpreted based on histology, with MeCP2 immunohistochemistry overlaid to assist interpretation. Cluster 1 corresponds to non-neoplastic pancreatic parenchyma; Cluster 2 to smooth muscle/mesenchyme; Cluster 3 to well-differentiated carcinoma with MeCP2 nuclear expression; Cluster 4 to well-differentiated carcinoma lacking MeCP2 expression; and Cluster 5 to poorly differentiated carcinoma. Genes consistently enriched in MeCP2-negative regions across two independent analyses are summarized in . ( b – e ) Pseudo– in situ hybridization (spatial feature plots) showing transcript distribution of ( b ) MeCP2, ( c ) TGFB1, ( d ) CDK6, and ( e ) VCAN. CDK6, and VCAN are predominantly expressed in MeCP2-negative clusters (4 and 5). Notably, VCAN-high stromal regions spatially correspond to MeCP2-positive CAFs, supporting a MeCP2-driven stromal remodeling program.

Article Snippet: Quantitative real-time PCR was performed on QuantStudio 1 Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, USA) using TaqMan Gene Expression Assays (Thermo Fisher Scientific, Waltham, MA, USA) for VCAN (Ref. No. Hs00171642_m1), TCF4 (Ref. No. Hs00162613_m1), and LEF1 (Ref. No. Hs01547250_m1).

Techniques: Immunohistochemistry, Expressing, In Situ Hybridization

Journal: Acta Histochemica et Cytochemica

Article Title: Role of MeCP2 in Shaping the Histopathological Heterogeneity of Ampullary Carcinoma

doi: 10.1267/ahc.25-00049

Figure Lengend Snippet: Immunohistochemical localization of VCAN in ampullary carcinoma. ( a , b ) VCAN immunostaining demonstrates epithelial and stromal expression in poorly differentiated adenocarcinoma. ( b , d ) MeCP2 immunostaining shows nuclear positivity in CAFs (red arrows) adjacent to carcinoma cells, whereas carcinoma cells are largely negative (green arrows). These findings indicate that VCAN-positive stromal fibroblasts correspond to MeCP2-positive CAFs. Bars = 20 μm.

Article Snippet: Quantitative real-time PCR was performed on QuantStudio 1 Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, USA) using TaqMan Gene Expression Assays (Thermo Fisher Scientific, Waltham, MA, USA) for VCAN (Ref. No. Hs00171642_m1), TCF4 (Ref. No. Hs00162613_m1), and LEF1 (Ref. No. Hs01547250_m1).

Techniques: Immunohistochemical staining, Immunostaining, Expressing

Journal: Acta Histochemica et Cytochemica

Article Title: Role of MeCP2 in Shaping the Histopathological Heterogeneity of Ampullary Carcinoma

doi: 10.1267/ahc.25-00049

Figure Lengend Snippet: Promoter structure of the VCAN gene. Schematic representation of the VCAN promoter region. A canonical TATA-box was located ~20 bp upstream of the transcription start site (TSS). Approximately 30 CpG dinucleotides (green) were distributed within a 500 bp region, although no typical CpG island was identified. Candidate TCF/LEF binding motifs were highlighted in yellow. Previously reported consensus motifs were designated as TCF/LEF #2 and #3, and a novel upstream sequence was predicted as TCF/LEF #1.

Article Snippet: Quantitative real-time PCR was performed on QuantStudio 1 Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, USA) using TaqMan Gene Expression Assays (Thermo Fisher Scientific, Waltham, MA, USA) for VCAN (Ref. No. Hs00171642_m1), TCF4 (Ref. No. Hs00162613_m1), and LEF1 (Ref. No. Hs01547250_m1).

Techniques: Binding Assay, Sequencing

Journal: Acta Histochemica et Cytochemica

Article Title: Role of MeCP2 in Shaping the Histopathological Heterogeneity of Ampullary Carcinoma

doi: 10.1267/ahc.25-00049

Figure Lengend Snippet: Functional analysis of TCF/LEF binding motifs and signaling pathway effects on VCAN expression. ( a , b ) Electrophoretic mobility shift assays (EMSAs) demonstrated a block-shift phenomenon at TCF/LEF #1 with both TCF4 and LEF1 antibodies (*), confirming protein-DNA interaction. By contrast, binding at motifs #2 and #3 was not evident. ( c ) Quantitative PCR showing VCAN mRNA expression in fibroblasts after LiCl treatment (0, 10, 20 mM). No induction was observed. ( d ) Quantitative PCR showing VCAN mRNA expression after PMA treatment (0, 10, 20 nM). No significant change was detected. ( e ) Forskolin treatment (0, 10, 20 μM) induced a dose-dependent upregulation of VCAN expression. ( f ) This forskolin-induced effect was not reproduced in VCAN-Luc promoter reporter assays.

Article Snippet: Quantitative real-time PCR was performed on QuantStudio 1 Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, USA) using TaqMan Gene Expression Assays (Thermo Fisher Scientific, Waltham, MA, USA) for VCAN (Ref. No. Hs00171642_m1), TCF4 (Ref. No. Hs00162613_m1), and LEF1 (Ref. No. Hs01547250_m1).

Techniques: Functional Assay, Binding Assay, Expressing, Electrophoretic Mobility Shift Assay, Blocking Assay, Real-time Polymerase Chain Reaction

Journal: Acta Histochemica et Cytochemica

Article Title: Role of MeCP2 in Shaping the Histopathological Heterogeneity of Ampullary Carcinoma

doi: 10.1267/ahc.25-00049

Figure Lengend Snippet: Detection of 5hmC in the VCAN promoter region. Base-resolution mapping of CpG sites upstream of the TCF/LEF #1 motif. 5hmC was consistently detected at the three most upstream CpG sites in all 10 of 10 samples, whereas it was detected in 4 of 10 samples at the next CpG and in 2 of 10 samples at the subsequent site.

Article Snippet: Quantitative real-time PCR was performed on QuantStudio 1 Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, USA) using TaqMan Gene Expression Assays (Thermo Fisher Scientific, Waltham, MA, USA) for VCAN (Ref. No. Hs00171642_m1), TCF4 (Ref. No. Hs00162613_m1), and LEF1 (Ref. No. Hs01547250_m1).

Techniques:

Journal: Acta Histochemica et Cytochemica

Article Title: Role of MeCP2 in Shaping the Histopathological Heterogeneity of Ampullary Carcinoma

doi: 10.1267/ahc.25-00049

Figure Lengend Snippet: MeCP2 binding to 5hmC-enriched CpG sites in the VCAN promoter. ( a ) Electrophoretic mobility shift assay (EMSA) using two segments (EMSA #1 and EMSA #2) of the 5hmC-enriched region. No significant protein-DNA binding was detected in the absence of 5hmC. ( b ) In the presence of 5hmC, distinct DNA-protein complexes were observed at both EMSA #1 and #2 (arrow). The addition of an anti-MeCP2 antibody induced a block-shift phenomenon (*), confirming specific MeCP2 binding to 5hmC-containing sequences.

Article Snippet: Quantitative real-time PCR was performed on QuantStudio 1 Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, USA) using TaqMan Gene Expression Assays (Thermo Fisher Scientific, Waltham, MA, USA) for VCAN (Ref. No. Hs00171642_m1), TCF4 (Ref. No. Hs00162613_m1), and LEF1 (Ref. No. Hs01547250_m1).

Techniques: Binding Assay, Electrophoretic Mobility Shift Assay, Blocking Assay

Journal: International Journal of Biological Sciences

Article Title: VCAN Is Essential for ERK5-Driven Tumorigenesis in Soft Tissue Sarcoma

doi: 10.7150/ijbs.121402

Figure Lengend Snippet: ERK5 and VCAN are involved in in vitro migration and adhesion, and in vivo tumor growth capabilities of SK-LMS-1 cells. (A) Wound healing assays were performed in SK-LMS-1 cells infected with lentiviruses carrying PLKO.1-shScramble (shSCR), PLKO.1-shRNA ERK5-1 (shERK5-1) or PLKO.1-shRNA VCAN-2 (shVCAN-2) vectors. Left panel shows the mean +/- SD of relative wound area of shSCR, shERK5-1 and shVCAN-2 SK-LM-S1 cells followed up to 30 hours, from 3 independent pools of infection. Right panel shows representative images from one experiment. (B) Percentage of shSCR, shERK5-1 and shVCAN-2 SK-LMS-1 cells fully adhered up to 330 minutes after seeding (left panel). Graphic represents the mean +/- SD of 3 independent experiments from different pools of infection. Right panels show representative images of cells taken 300 minutes after seeding. Blue arrows mark fully adherent and expanded cells, red arrows mark not attached cells. (C) Left panel: tumor growth of 2 × 10 6 subcutaneously inoculated shSCR, shERK5-1 and shVCAN-2 SK-LM-S1 cells in NSG mice (n=4). The graphic represents the mean ± SEM. Right panel: images of excised tumors. The unpaired Student's t-test was used to assess statistical significance. *p < 0.05; **p < 0.01; ***p < 0.001 .

Article Snippet: For shRNA assays, all plasmids, with a PLKO.1 basis, were purchased from Merck (Tres Cantos, Madrid, Spain): shRNA ERK5-1 Human (TRCN0000010275), shRNA ERK5-2 Human (TRCN0000197264), shRNA VCAN-1 Human (TRCN0000033637), shRNA VCAN-2 Human (TRCN0000033638), and for mouse cell lines shRNA ERK5 (TRCN0000232396), and shRNA VCAN (TRCN0000175477).

Techniques: In Vitro, Migration, In Vivo, Infection, shRNA