human gbm cell lines ln229  (ATCC)

Journal: MedComm

Article Title: Integrative Single‐Cell Analysis Reveals Targetable Vacuole Membrane Protein 1‐Mediated Mechanism of Tumor Angiogenesis in Glioblastoma

doi: 10.1002/mco2.70619

Figure Lengend Snippet: Overexpression of VMP1 promotes tumor growth. (A) Representative western blot image validating exogenous overexpression of VMP1 in U87 and U251 cell lines. (B) U87 subcutaneous xenografts of overexpressed VMP1 (VMP1‐OE) and control vector at Day 17 post‐injection ( n = 9). (C) The weight of the tumor grafts. (D) Quantification of normalized tumor weights of VMP1‐OE and vector (measured every 4 days from Day 3) ( n = 9). (E) Bioluminescence imaging of U87 VMP1‐OE and vector orthotopic xenografts at different time points from Day 3 to Day 17 ( n = 6). (F) Representative images of hematoxylin and eosin‐stained sections at Day 17 post‐injection. Tumor is indicated within the dashed line. Scale bar, 1000 µm. (G) Relative total photon flux of bioluminescence in mice with U87 VMP1‐OE and vector. (H) Kaplan–Meier survival analysis of mice with U87 VMP1‐OE and vector intracranial xenografts ( n = 6). (I) Top, immunohistochemical staining showing Ki67‐positive cells in subcutaneous and intracranial xenografts. Scale bar, 100 µm. Bottom, quantification of Ki67‐positive cells (%) in subcutaneous and intracranial models. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Human embryonic kidney cells 293T (293T) and human GBM cell lines U87 and U251 were purchased from the American Type Culture Collection (ATCC).

Techniques: Over Expression, Western Blot, Control, Plasmid Preparation, Injection, Imaging, Staining, Immunohistochemical staining

Journal: MedComm

Article Title: Integrative Single‐Cell Analysis Reveals Targetable Vacuole Membrane Protein 1‐Mediated Mechanism of Tumor Angiogenesis in Glioblastoma

doi: 10.1002/mco2.70619

Figure Lengend Snippet: VMP1 promoted tumor growth was independent of autophagy. (A) Representative western blot images of autophagy markers (p62 and LC3 I/II) in U87 and U251 cell lines with VMP1‐OE. (B) Representative transmission electron microscopy images of a cell in U87 and U251 with VMP1‐OE, showing no differences in autophagosome formation. Top: scale bar, 2 µm. Bottom: scale bar, 500 nm. (C) Western blot images of tissue samples from our glioma cohort (glioma) ( n = 47) and normal brain tissue (N), showing the protein expression of autophagy markers (p62, Beclin 1, and LC3 I/II). (D) Quantification of western blot images, patients were separated into two groups based on median VMP1 expression: VMP1 low glioma ( n = 23) and VMP1 high glioma ( n = 24). (E) Correlation analysis of western blot quantification value between VMP1 and autophagy markers (p62, Beclin 1, and LC3 I/II). (F) Confirmation of VMP1 knockdown in U87 and U251 cells using two different targeting sequences by western blot analysis. (G) U87 subcutaneous xenografts of VMP1 knockdown (shVMP1) and control vector (shNC) at Day 27 post‐injection ( n = 10) (left), and the tumor volume measured from Day 14 to Day 27 (right). (H) Bioluminescence imaging of U87 shVMP1 and vector orthotopic xenografts at Day 28. (I) Representative images of hematoxylin and eosin‐stained sections at Day 28 post‐injection. Scale bar, 2000 µm. (J) Kaplan–Meier survival analysis of mice with U87 shVMP1 and vector intracranial xenografts ( n = 5). (K) Representative western blot images of U87 shVMP1 and vector subcutaneous xenografts showing the expression of autophagy markers p62 and LC3 I/II (left). Quantification of band intensities normalized to GAPDH (right). ns, no statistical significance; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Human embryonic kidney cells 293T (293T) and human GBM cell lines U87 and U251 were purchased from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Transmission Assay, Electron Microscopy, Expressing, Knockdown, Control, Plasmid Preparation, Injection, Imaging, Staining

Journal: MedComm

Article Title: Integrative Single‐Cell Analysis Reveals Targetable Vacuole Membrane Protein 1‐Mediated Mechanism of Tumor Angiogenesis in Glioblastoma

doi: 10.1002/mco2.70619

Figure Lengend Snippet: VMP1 mediates angiogenesis and vascular permeability through activation of endothelial cells in the TME. (A) Representative western blot image (top) and quantification (bottom) of VEGFR2 expression in human primary endothelial cells (HUVEC) after culturing with conditioned medium (CM) collected from VMP1‐overexpressing glioblastoma cell lines U87 and U251. (B) Representative immunofluorescence image of VEGFR2 expression (red) and DAPI (blue) in HUVEC cultured with conditioned medium. Scale bar, 200 µm. (C) Representative immunofluorescence staining image of VE‐cadherin expression (red) and DAPI (blue) in HUVEC with CM. Scale bar, 200 µm. (D) Human protein angiogenesis array showing 55 angiogenesis‐related proteins in the CM collected. (E) Quantification of eight of the angiogenesis‐related proteins, including tissue factor (TF), granulocyte‐macrophage colony stimulating factor (GM‐CSF), macrophage inflammatory protein 1α (MIP1α), Serpin E1, Thrombospondin‐1 (THBS1), Angiogenin, tissue inhibitor of metalloproteinase 1 (TIMP‐1), and VEGF‐C. (F) Spatial distribution of spot degree between VMP1 high cancer cells and endothelial cells in the spatial mRNA dataset. (G–I) Spatial distribution of angiogenesis (G), Serpin E1 (H), and TIMP1 (I) expression in the spatial mRNA dataset. * p < 0.05; ** p < 0.01; *** p <0 .001.

Article Snippet: Human embryonic kidney cells 293T (293T) and human GBM cell lines U87 and U251 were purchased from the American Type Culture Collection (ATCC).

Techniques: Permeability, Activation Assay, Western Blot, Expressing, Immunofluorescence, Cell Culture, Staining

Journal: MedComm

Article Title: Integrative Single‐Cell Analysis Reveals Targetable Vacuole Membrane Protein 1‐Mediated Mechanism of Tumor Angiogenesis in Glioblastoma

doi: 10.1002/mco2.70619

Figure Lengend Snippet: Targeted inhibition of VEGFA represses VMP1‐mediated tumor growth. (A) The treatment timeline and bioluminescence detection of mice treated with bevacizumab (BEV) at different time points (Day 3, Day 7, and Day 13). (B) Bioluminescence detection of U87 tumor‐bearing mice treated with BEV and vehicle. (C) Hematoxylin and eosin staining of mice brain, dotted area indicates the tumor region. Scale bar, 1000 µm. (D) Relative total photon flux in orthotopic mice model after treatment. (E) Changes in body weight in mice after treatments. (F) Kaplan–Meier survival of mice after being treated with bevacizumab and vector control. ns, no statistical significance.

Article Snippet: Human embryonic kidney cells 293T (293T) and human GBM cell lines U87 and U251 were purchased from the American Type Culture Collection (ATCC).

Techniques: Inhibition, Staining, Plasmid Preparation, Control

Journal: Oncology Reports

Article Title: S100B drives glioblastoma invasion and migration through TGF-β2-mediated epithelial-mesenchymal transition

doi: 10.3892/or.2025.9025

Figure Lengend Snippet: Lentiviral transduction-induced knockdown of S100B in the LN229 glioblastoma multiforme cell line. (A) Detection of S100B expression in LN229 cells by flow cytometry. (B) Representative immunofluorescence images of S100B. Scale bars, 50 µm. (C) LN229 glioma cells were transduced with shS100B and negative control vectors. (D and E) mRNA and protein levels of S100B were analyzed by reverse transcription-quantitative PCR and western blotting. Data are presented as the mean ± SEM. (F) S100B expression in NC and shS100B groups was measured by immunofluorescence staining. DAPI represents cell nuclei. Scale bar, 50 µm. The statistical analyses of the relative mean fluorescence intensity and the relative percentage of positive cells were conducted. Data are presented as the mean ± SEM. **P<0.01 and ****P<0.0001. shS100B, shRNA S100B; NC, negative control.

Article Snippet: The human GBM cell line LN229 (cat. no. CRL-2611) was obtained from the American Type Culture Collection.

Techniques: Transduction, Knockdown, Expressing, Flow Cytometry, Immunofluorescence, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Staining, Fluorescence, shRNA

Journal: Oncology Reports

Article Title: S100B drives glioblastoma invasion and migration through TGF-β2-mediated epithelial-mesenchymal transition

doi: 10.3892/or.2025.9025

Figure Lengend Snippet: S100B affects the proliferation of LN229 cells and tumor growth in vivo . (A) Colony formation assay assessed the proliferative capacity of NC and shS100B groups, Scale bar, 500 µm. (B) DNA synthesis ability was assessed using the EdU assays. Scale bar, 50 µm. (C) Cell Counting Kit-8 assay of proliferation in NC and shS100B LN229 glioblastoma multiforme cells. (D and E) Subcutaneous tumor model, tumor size and volume. (F) Representative immunofluorescence staining images of S100B in tumor tissue sections. DAPI represents cell nuclei. Statistical analysis of relative mean fluorescence intensity of S100B was performed. Data are presented as the mean ± SEM. Scale bar, 100 µm. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. shS100B, shRNA S100B; NC, negative control.

Article Snippet: The human GBM cell line LN229 (cat. no. CRL-2611) was obtained from the American Type Culture Collection.

Techniques: In Vivo, Colony Assay, DNA Synthesis, Cell Counting, Immunofluorescence, Staining, Fluorescence, shRNA, Negative Control

Journal: Oncology Reports

Article Title: S100B drives glioblastoma invasion and migration through TGF-β2-mediated epithelial-mesenchymal transition

doi: 10.3892/or.2025.9025

Figure Lengend Snippet: S100B affects the invasion and migration of GBM cells via epithelial-mesenchymal transition. (A) Detection of cell migration ability by cross-scratch assay; Scale bar, 500 µm. Cell migration rate = Cell migration area/cross scratch area. (B and C) Transwell assays analyzed the migratory and invasive capacity of NC and shS100B GBM cells, Scale bar, 200 µm. Relative proportion of migratory or invasive cells: Number of migratory or invasive cells in shS100B group/number of migratory or invasive cells in NC group. (D) Volcano plot showing the DEGs in the NC and shS100B groups. (E) GO enrichment analysis of the functions of downregulated DEG. (F) E-cadherin, N-cadherin and vimentin expression in NC and shS100B LN229 cells were measured by immunofluorescence staining. Blue represents cell nuclei, red represents E-cadherin, N-cadherin and vimentin. Statistical analysis of relative mean fluorescence intensity was performed. Scale bar, 50 µm. Data are presented as the mean ± SEM. **P<0.01 and ****P<0.0001. GBM, glioblastoma multiforme; shS100B, shRNA S100B; NC, negative control; DEGs, differentially expressed genes; GO, Gene Ontology.

Article Snippet: The human GBM cell line LN229 (cat. no. CRL-2611) was obtained from the American Type Culture Collection.

Techniques: Migration, Wound Healing Assay, Expressing, Immunofluorescence, Staining, Fluorescence, shRNA, Negative Control

Journal: Oncology Reports

Article Title: S100B drives glioblastoma invasion and migration through TGF-β2-mediated epithelial-mesenchymal transition

doi: 10.3892/or.2025.9025

Figure Lengend Snippet: TGF-β2 induces epithelial-mesenchymal transition and enhances the invasion and migration of glioblastoma multiforme cells. (A) Relevant downregulated genes of the mesenchyme morphogenesis pathway. (B and C) Expression of TGF-β2 in NC and shS100B LN229 cells was analyzed by reverse transcription-quantitative PCR and western blotting. (D) Cross-scratch assay evaluated the migratory capacity of NC, shS100B and shS100B + TGF-β2 groups, Scale bar, 500 µm. (E) Transwell assay analyzed the migratory or invasive capacity of NC, shS100B and shS100B + TGF-β2 groups; Scale bar, 200 µm. Relative proportion of migratory or invasive cells: Number of migratory or invasive cells in shS100B group or shS100B + TGF-β2 group/number of migrated or invaded cells in NC group. (F) E-cadherin, N-cadherin and vimentin were analyzed by immunofluorescence staining. Blue represents cell nuclei, and red represents E-cadherin, N-cadherin and vimentin. Scale bar, 50 µm. Data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. NC, negative control; shS100B, shRNA S100B.

Article Snippet: The human GBM cell line LN229 (cat. no. CRL-2611) was obtained from the American Type Culture Collection.

Techniques: Migration, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Wound Healing Assay, Transwell Assay, Immunofluorescence, Staining, Negative Control, shRNA