Journal: Cancer Cell International

Article Title: Hsa_circ_0059511 promote glioma cell proliferation and migration through hsa-miR-194-5p/ HBEGF axis

doi: 10.1186/s12935-025-03815-w

Figure Lengend Snippet: HBEGF was a target of hsa-miR-194-5p. A The Venn diagram displayed the number of hsa-miR-194-5p target genes predicted by different databases and shared target genes from three databases. B The Venn diagram showed the number of shared genes between hsa-miR-194-5p target genes and DEGs. C The lollipop plot showed the rank of target scores for the top 10 target genes from predicting results. D The binding sites between hsa-miR-194-5p and HBEGF. E Pearson correlation analysis was used to explore the relationship between the expression levels of HBEFG and hsa-miR-194-5p. F , G The quantification of relative luciferase activity in T98G and U251 cells by the dual-luciferase reporter gene system. H , I The quantification of relative hsa-miR-194-5p mRNA levels in T98G and U251 cells by the RIP assay. * p < 0.05, ** p < 0.05, *** p < 0.001

Article Snippet: Cells were incubated overnight at 4 °C with a rabbit monoclonal anti-HBEGF antibody (Cell Signaling Technology, Danvers, MA, USA) at a 1:1000 dilution in phosphate-buffered saline containing 1% bovine serum albumin.

Techniques: Binding Assay, Expressing, Luciferase, Activity Assay

Journal: Cancer Cell International

Article Title: Hsa_circ_0059511 promote glioma cell proliferation and migration through hsa-miR-194-5p/ HBEGF axis

doi: 10.1186/s12935-025-03815-w

Figure Lengend Snippet: The prognosis of patients with glioma was impacted by the upregulation of HBEGF expression in glioma samples. A The quantification of HBEGF expression levels in glioma tissues by five expression profiling by array datasets. B The quantification of HBEGF expression levels in glioma tissues by five expression profiling by high throughput sequencing datasets. C Representative images of HBEGF expression levels in cells by western blot. D Representative images of HBEGF expression levels in glioma tissues by western blot. E , F The quantification of HBEGF expression levels by western blot assay. G – O The Kaplan–Meier curves showed the effect of HBEGF expression levels on the prognosis of patients with glioma. * p < 0.05, ** p < 0.05, *** p < 0.001

Article Snippet: Cells were incubated overnight at 4 °C with a rabbit monoclonal anti-HBEGF antibody (Cell Signaling Technology, Danvers, MA, USA) at a 1:1000 dilution in phosphate-buffered saline containing 1% bovine serum albumin.

Techniques: Expressing, Next-Generation Sequencing, Western Blot

Journal: Cancer Cell International

Article Title: Hsa_circ_0059511 promote glioma cell proliferation and migration through hsa-miR-194-5p/ HBEGF axis

doi: 10.1186/s12935-025-03815-w

Figure Lengend Snippet: Overexpression of HBEGF recovered hsa-miR-194-5p-blocked glioma cell proliferation, migration, and invasion. A , B The quantification of HBEGF expression levels in T98G and U251 cells by qRT-PCR. C , D The quantification of proliferation rate of T98G and U251 cells by MTT assay. E Representative images of T98G and U251 cell invasion experiments. F , G The quantification of invasion rate of T98G and U251 cells by the Transwell assay. H , I The quantification of proliferation rate of T98G and U251 cells by Edu assay. J Representative images of T98G and U251 cell proliferation experiment. *p < 0.05, **p < 0.05, ***p < 0.001

Article Snippet: Cells were incubated overnight at 4 °C with a rabbit monoclonal anti-HBEGF antibody (Cell Signaling Technology, Danvers, MA, USA) at a 1:1000 dilution in phosphate-buffered saline containing 1% bovine serum albumin.

Techniques: Over Expression, Migration, Expressing, Quantitative RT-PCR, MTT Assay, Transwell Assay, EdU Assay

Journal: Cancer Cell International

Article Title: Hsa_circ_0059511 promote glioma cell proliferation and migration through hsa-miR-194-5p/ HBEGF axis

doi: 10.1186/s12935-025-03815-w

Figure Lengend Snippet: Hsa_circ_0059511 facilitated the expression of HBEGF via sponging hsa-miR-194-5p. A , B Representative images of the HBEGF immunofluorescence staining in T98G and U251 cells. C The quantification of the HBEGF expression levels in T98G cell by qRT-PCR. D Representative images of the HBEGF expression levels in T98G and U251 cells by western blot. E , F The quantification of the HBEGF expression levels in T98G and U251 cells by western blot assay. *p < 0.05, **p < 0.05, ***p < 0.001

Article Snippet: Cells were incubated overnight at 4 °C with a rabbit monoclonal anti-HBEGF antibody (Cell Signaling Technology, Danvers, MA, USA) at a 1:1000 dilution in phosphate-buffered saline containing 1% bovine serum albumin.

Techniques: Expressing, Immunofluorescence, Staining, Quantitative RT-PCR, Western Blot

Journal: Cancer Cell International

Article Title: Hsa_circ_0059511 promote glioma cell proliferation and migration through hsa-miR-194-5p/ HBEGF axis

doi: 10.1186/s12935-025-03815-w

Figure Lengend Snippet: Knockdown of Hsa_circ_0059511 was reduced in nude mice. A Representative images of subcutaneous tumorigenesis induced by injecting WT U251 cells and U251 cells that express shRNA NC and shRNA hsa_circ_0059511 are stable. B The effects of the hsa_circ_0059511 knockdown on the volume of the neoplasms. C The effects of the hsa_circ_0059511 knockdown on the weight of the neoplasms. D The representative images of HE staining of neoplasms, hyperbasophilic cells ( black arrow ), hyperchromatic cells ( red arrow ), vessel proliferation ( yellow arrow ), mitosis ( blue arrow ). E The representative images of immunohistochemical (IHC) staining of glioma tissues, HBEGF-positive regions in the cytoplasm of tumor cells ( blue arrow ) in glioma tissue were more abundant than in normal brain tissue. F The quantification of HBEGF-positive regions in glioma tissue and normal brain tissue. G The representative images of IHC staining of neoplasms, the HBEGF positivity and positive area in the cytoplasm of the cells ( blue arrow ) in the hsa_circ_0059511 knockdown group were significantly lower than those in the WT group and the normal control group. H The quantification of the HBEGF positive rate in neoplasms. I The quantification of the expression levels of hsa-cicr-0059511 in the WT group, the normal control group, and the hsa_circ_0059511 knockdown group by qRT-PCR. J The quantification of the expression levels of hsa-miR-194-5p in the WT group, the normal control group, and the hsa_circ_0059511 knockdown group by qRT-PCR. K An overview of the mechanism by which hsa_circ_0059511 promotes the development of glioma. * p < 0.05, ** p < 0.05, *** p < 0.001

Article Snippet: Cells were incubated overnight at 4 °C with a rabbit monoclonal anti-HBEGF antibody (Cell Signaling Technology, Danvers, MA, USA) at a 1:1000 dilution in phosphate-buffered saline containing 1% bovine serum albumin.

Techniques: Knockdown, shRNA, Staining, Immunohistochemical staining, Immunohistochemistry, Control, Expressing, Quantitative RT-PCR

Journal: Proteomics

Article Title: HB-EGF-loaded nanovesicles enhance trophectodermal spheroid attachment and invasion.

doi: 10.1002/pmic.202200145

Figure Lengend Snippet: FIGURE 1 Production and characterisation of NVHBEGF. (A) NVHBEGF were generated by serial extrusion (10, 5, 1 μm filters, 13 times per membrane) of human trophectodermal cells (T3-TSCs) with either 50 ng/mL of HB-EGF or PBS and purified using density-cushion ultracentrifugation to obtain 7 fractions (F1-7) of increasing density. NV-containing fraction (F5) was obtained. (B) Cryo-electron microscopic image of NVHBEGF displayed spherical and morphologically intact structures; scale 100 nm. (C) Size distribution of NVHBEGF based on cryo-electron microscopic images (n = 4) reveal enrichment of particles 50–150 nm in diameter. (D) Abundance of HB-EGF using mass spectrometry analysis; normalized LFQ intensities (log2) of HB-EGF between NVHBEGF and NVs generated using the same workflow from hTSCs and mouse embryonic fibroblasts. Additional marker proteins associated with EGFR, HSPG2, and CD44 were also compared, noting similarly expression in both NV groups. (E) Western blot display of HB-EGF enrichment in NVHBEGF compared to NVs (n = 3). NVHBEGF, HB-EGF-loaded NVs; NVs, nanovesicles.

Article Snippet: Primary antibodies used include mouse monoclonal against CD44 (#119863, Abcam), and HB-EGF (#27450, Cell Signaling Tech- nology).

Techniques: Generated, Membrane, Purification, Mass Spectrometry, Marker, Expressing, Western Blot

Journal: Proteomics

Article Title: HB-EGF-loaded nanovesicles enhance trophectodermal spheroid attachment and invasion.

doi: 10.1002/pmic.202200145

Figure Lengend Snippet: FIGURE 2 Uptake of NVHBEGF and NVs by HEC1A endometrial cells. (A) Confocal fluorescent microscopy images demonstrating uptake of NVHBEGF or NVs labelled with DiI lipophilic fluorescent dye labelled (red) by HEC1A endometrial cells after a 2-h incubation (n = 3). (B) Fluorescent Z-stack image displaying intracellular distribution of DiI-labelled NVHBEGF (red). Nuclei of HEC1A endometrial cells were stained with Hoechst (blue). Scale bar 10 μm. NVHBEGF, HB-EGF-loaded NVs; NVs, nanovesicles.

Article Snippet: Primary antibodies used include mouse monoclonal against CD44 (#119863, Abcam), and HB-EGF (#27450, Cell Signaling Tech- nology).

Techniques: Microscopy, Incubation, Staining

Journal: Proteomics

Article Title: HB-EGF-loaded nanovesicles enhance trophectodermal spheroid attachment and invasion.

doi: 10.1002/pmic.202200145

Figure Lengend Snippet: FIGURE 3 NVHBEGF remodel the phosphoproteome landscape in HEC1A endometrial cells. (A) Workflow for NVHBEGF and NV treatment onto recipient HEC1A endometrial cells, including a 2-step treatment of erlotinib (EGFR inhibition) followed by NVHBEGF stimulation, and subsequent cell phosphoproteome preparation and analysis. (B) Heatmap expression (log2) of phosphorylated proteins and phosphosites of players of the EGFR signalling pathway, which are downregulated when EGFR is inhibited by erlotinib (white corresponds to missing values). (C) (Top) Erlotinib inhibited the phosphorylation of 421 proteins (compared to PBS), while subsequent NVHBEGF treatment induced phosphorylation of 261 of the inhibited proteins; (Bottom) bubble plot displaying key biological processes and pathways corresponding to the 421 and 261 proteins respectively. NVHBEGF, HB-EGF-loaded NVs; NVs, nanovesicles.

Article Snippet: Primary antibodies used include mouse monoclonal against CD44 (#119863, Abcam), and HB-EGF (#27450, Cell Signaling Tech- nology).

Techniques: Inhibition, Expressing, Phospho-proteomics

Journal: Proteomics

Article Title: HB-EGF-loaded nanovesicles enhance trophectodermal spheroid attachment and invasion.

doi: 10.1002/pmic.202200145

Figure Lengend Snippet: FIGURE 4 NVHBEGF remodel the proteome landscape and EGFR signaling network at the time of implantation. (A) Workflow employed for proteomic analysis of stimulated HEC1A endometrial cells. (B) Proteins uniquely identified and significantly upregulated in NVHBEGF- or NV-treated HEC1A cells compared to PBS. (C) Pre-treatment of HEC1A cells with erlotinib followed by NVHBEGF downregulated the expression of 127 proteins compared to NVHBEGF, which are categorised into related biological processes. (D) NVHBEGF- and ErloNVHBEGF–mediated phosphorylation levels of 13 kinases that are matched to downregulated proteins. (E) Comparative analysis of HEC1A cellular proteome treated with NVHBEGF compared to ErloNVHBEGF and PBS, and a two-way scatter plot highlighting top dysregulated proteins in the presence of EGFR inhibitor, erlotinib. (F) Bubble plot display of biological processes and pathways associated with proteins significantly upregulated (including unique) by NVHBEGF treatment and proteins significantly downregulated (including not detected) in NVHBEGF compared to ErloNVHBEGF and PBS. NVHBEGF, HB-EGF-loaded NVs; NVs, nanovesicles.

Article Snippet: Primary antibodies used include mouse monoclonal against CD44 (#119863, Abcam), and HB-EGF (#27450, Cell Signaling Tech- nology).

Techniques: Expressing, Phospho-proteomics

Journal: Proteomics

Article Title: HB-EGF-loaded nanovesicles enhance trophectodermal spheroid attachment and invasion.

doi: 10.1002/pmic.202200145

Figure Lengend Snippet: FIGURE 5 NVHBEGF enhances attachment to endometrial cells and outgrowth and invasion in Matrigel of trophectodermal spheroids. (A) Experimental workflow for co-culture attachment assay. (B) Box plot indicating percentage of spheroid attachment to HEC1A endometrial cells following treatment with PBS, NVHBEGF, NV, HB-EGF, or ErloNVHBEGF (n = 5), where rate of spheroid attachment (%) is the number of attached spheroids divided by the number of seeded spheroids expressed as a percentage. (C) Experimental workflow for TSC spheroid outgrowth and invasion into Matrigel. (D) Box plot indicating quantified area of TSC spheroid outgrowth and invasion into Matrigel 72 h following treatment with PBS, NVHBEGF, NV, HB-EGF, or ErloNVHBEGF (n = 8). (E) Bright-field microscopic images of TSC spheroids outgrowth and invasion into Matrigel 72 h following treatment with PBS, NVHBEGF, NV, HB-EGF, or ErloNVHBEGF. Scale bar 100 μm. (F) Area of outgrowth extending from spheroid taken for measurements is shaded in grey and quantified using ImageJ. *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.001.

Article Snippet: Primary antibodies used include mouse monoclonal against CD44 (#119863, Abcam), and HB-EGF (#27450, Cell Signaling Tech- nology).

Techniques: Co-Culture Assay