Journal: Journal of Advanced Research

Article Title: S100B induces angiogenesis via the clathrin/FOXO1/β-catenin signaling pathway and contributes to Bevacizumab resistance in epithelial ovarian cancer

doi: 10.1016/j.jare.2025.05.060

Figure Lengend Snippet: Flowchart of the study. First, BEV-sensitive and BEV-resistant ovarian cancer mouse models were constructed, and RNA sequencing was performed on tumor tissue. Then, a BEV-related prognostic signature was established using machine learning, and S100B was identified as the most important molecule regulating BEV sensitivity in ovarian cancer. Its function and mechanism were analyzed in vitro. Finally, BEV efficacy when combined with an S100B inhibitor was verified in vivo. BEV: Bevacizumab; DEGs: Differentially expressed genes.

Article Snippet: During phenotype experiments with HUVECs, recombinant S100B protein (MCE, HY- P70659 ) was added at concentrations of 10, 100, and 1000 mg/mL.

Techniques: Construct, RNA Sequencing, In Vitro, In Vivo

Journal: Journal of Advanced Research

Article Title: S100B induces angiogenesis via the clathrin/FOXO1/β-catenin signaling pathway and contributes to Bevacizumab resistance in epithelial ovarian cancer

doi: 10.1016/j.jare.2025.05.060

Figure Lengend Snippet: Screening S100B as the main molecule regulating the sensitivity of ovarian cancer to BEV. (A) Kaplan–Meier survival analysis of OS (left) and PFS (right) of patients with high S100B expression treated with chemotherapy with or without BEV (from the GSE140082 dataset). (B) Kaplan–Meier survival analysis of OS (left) and PFS (right) of patients with low S100B expression treated with chemotherapy with or without BEV (from the GSE140082 dataset). (C) Western blot analysis of S100B protein expression in tumor tissue from BEV-sensitive and BEV-resistant mice. (D) Western blot analysis of S100B protein levels in mouse tumor tissue. (E) Immunohistochemical staining of S100B protein in BEV-sensitive and BEV-resistant mice. (F) Statistical histochemical staining of S100B protein in mouse tumor tissue. BEV:Bevacizumab; **: P < 0.01.

Article Snippet: During phenotype experiments with HUVECs, recombinant S100B protein (MCE, HY- P70659 ) was added at concentrations of 10, 100, and 1000 mg/mL.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Journal of Advanced Research

Article Title: S100B induces angiogenesis via the clathrin/FOXO1/β-catenin signaling pathway and contributes to Bevacizumab resistance in epithelial ovarian cancer

doi: 10.1016/j.jare.2025.05.060

Figure Lengend Snippet: Co-culture of S100B overexpressing ovarian cancer cells promotes HUEVC angiogenesis and migration. (A) Representative Western blot images and their densitometric quantification showing comparative S100B protein expression profiles among the three ovarian cancer cell lines (A2780, HEY, and OVCAR3) (B) Western blot analysis and statistical analysis of OVCAR3 and HEY cells overexpressing S100B. RT-qPCR analysis of alternative angiogenic factor mRNA expression levels in (C) OVCAR3 and (D) HEY ovarian cancer cell lines following S100B overexpression. (E) The level of S100B in the supernatant of OVCAR3 and HEY cells overexpressing S100B significantly increased, as determined by ELISA. (F) There was no significant change in VEGFA levels in the supernatant of OVCAR3 and HEY cells overexpressing S100B. (G, H) HUVEC tube formation significantly increased after co-culture with ovarian cancer cell lines overexpressing S100B, with or without exogenous BEV. (I, J) Co-culture with ovarian cancer cells overexpressing S100B significantly promoted HUVEC sprouting with or without exogenous BEV. (K, L) Co-culture of ovarian cancer cells overexpressing S100B significantly promoted migration of HUVECs in a Transwell system with or without exogenous BEV. (M, N) Co-culture of ovarian cancer cell lines overexpressing S100B significantly promoted healing of HUVECs with or without exogenous BEV in a scratch assay. BEV: Bevacizumab; NC: negative control; OE: overexpression; *: P < 0.05; **: P < 0.01; ***: P < 0.001; ****: P < 0.0001. Scale: All panels are 50 μm.

Article Snippet: During phenotype experiments with HUVECs, recombinant S100B protein (MCE, HY- P70659 ) was added at concentrations of 10, 100, and 1000 mg/mL.

Techniques: Co-Culture Assay, Migration, Western Blot, Expressing, Quantitative RT-PCR, Over Expression, Enzyme-linked Immunosorbent Assay, Wound Healing Assay, Negative Control

Journal: Journal of Advanced Research

Article Title: S100B induces angiogenesis via the clathrin/FOXO1/β-catenin signaling pathway and contributes to Bevacizumab resistance in epithelial ovarian cancer

doi: 10.1016/j.jare.2025.05.060

Figure Lengend Snippet: Exogenous recombinant S100B protein promoted vascular formation and migration of HUEVCs. Different concentrations of recombinant S100B protein, with or without BEV, promoted (A, B) angiogenesis in a tube formation assay, (C, D) sprouting in a fibrin bead sprouting assay, (E, F) migration in a Transwell assay, and (G, H) healing of endothelial cells in a wound healing assay. (I) Tip cell marker expression in HUVECs significantly increased by rt-qPCR detection after treatment with recombinant S100B protein. BEV: Bevacizumab; *: P < 0.05; **: P < 0.01; ***: P < 0.001; ****: P < 0.0001. Scale: All panels are 50 μm.

Article Snippet: During phenotype experiments with HUVECs, recombinant S100B protein (MCE, HY- P70659 ) was added at concentrations of 10, 100, and 1000 mg/mL.

Techniques: Recombinant, Migration, Tube Formation Assay, Transwell Assay, Wound Healing Assay, Marker, Expressing, Quantitative RT-PCR

Journal: Journal of Advanced Research

Article Title: S100B induces angiogenesis via the clathrin/FOXO1/β-catenin signaling pathway and contributes to Bevacizumab resistance in epithelial ovarian cancer

doi: 10.1016/j.jare.2025.05.060

Figure Lengend Snippet: S100B enters endothelial cells through clathrin-dependent endocytosis to promote angiogenesis and migration. (A) Western blot detection showed that there was no significant change in RAGE receptor expression in endothelial cells after exogenous S100B treatment. (B) Western blot detection showed that protein levels of S100B in endothelial cells significantly increased after co-culture with ovarian cancer cells overexpressing S100B. (C) S100B total protein and nuclear protein in endothelial cells significantly increased after treatment with exogenous S100B, as determined by western blot analysis. (D) Western blot analysis of protein levels of S100B in endothelial cells after pretreatment with the clathrin-mediated endocytosis inhibitor Pitstop-2 or caveolae/caveolin-1-mediated endocytosis inhibitor nystatin. Changes in the (F, G) tubular phenotype, (H, I) sprouting phenotype, (J, K) migration phenotype (Transwell assay), and (L–M) wound healing phenotype (scratch migration assay) of endothelial cells after treatment with the RAGE receptor inhibitor FPS-ZM1, the clathrin-mediated endocytosis inhibitor Pitstop-2, and the caveolae/caveolin-mediated endocytosis inhibitor nystatin. BEV: Bevacizumab; *: P < 0.05; **: P < 0.01; ***: P < 0.001; ****: P < 0.0001. Scale: All panels are 50 μm.

Article Snippet: During phenotype experiments with HUVECs, recombinant S100B protein (MCE, HY- P70659 ) was added at concentrations of 10, 100, and 1000 mg/mL.

Techniques: Migration, Western Blot, Expressing, Co-Culture Assay, Transwell Assay

Journal: Journal of Advanced Research

Article Title: S100B induces angiogenesis via the clathrin/FOXO1/β-catenin signaling pathway and contributes to Bevacizumab resistance in epithelial ovarian cancer

doi: 10.1016/j.jare.2025.05.060

Figure Lengend Snippet: S100B promotes tube formation and migration of endothelial cells through the FOXO1/β-catenin signaling pathway. (A) DIA proteomics thermogram of differentially expressed proteins in HUVECs with or without exogenous S100B. (B) Function and pathway enrichment analysis by Metascape database of differentially expressed proteins. (C–E) Western blot analysis showed that levels of S100B in ovarian cancer tissue of BEV-resistant mice significantly increased, while FOXO1 levels decreased. (F) Histogram of an RT-qPCR analysis of downstream transcriptional target genes of β-catenin. (G–J) Western blot analysis showed that FOXO1 total protein and nuclear protein levels in endothelial cells significantly decreased after treatment with exogenous S100B, while levels of β-catenin and MMP7 significantly increased. (K–M) Western blot analysis of FOXO1 and β-catenin levels in S100B-treated endothelial cells overexpressing FOXO1 compared with the control group. Endothelial cells transfected with a control virus or FOXO1 -overexpressing lentivirus and treated with S100B were evaluated for their (N) tubular phenotype by tube formation assay, (O) sprouting phenotype by fibrin bead sprouting assay, and (P) migration phenotype by Tranwell assay. (Q) Schematic diagram of the mechanism of exogenous S100B on endothelial cell angiogenesis. *: P < 0.05; **: P < 0.01;^: P < 0.05 vs. S100B−/oeFOXO1−; #: P < 0.05 vs. S100B−/oeFOXO1+; &: P < 0.05 vs. S100B+/oeFOXO1 − . Scale: All panels are 50 μm.

Article Snippet: During phenotype experiments with HUVECs, recombinant S100B protein (MCE, HY- P70659 ) was added at concentrations of 10, 100, and 1000 mg/mL.

Techniques: Migration, Western Blot, Quantitative RT-PCR, Control, Transfection, Virus, Tube Formation Assay

Journal: Journal of Advanced Research

Article Title: S100B induces angiogenesis via the clathrin/FOXO1/β-catenin signaling pathway and contributes to Bevacizumab resistance in epithelial ovarian cancer

doi: 10.1016/j.jare.2025.05.060

Figure Lengend Snippet: The S100B inhibitor pentamidine in combination with BEV improves the ovarian cancer response compared to BEV alone. (A) Flowchart of the in vivo experiment. Two weeks after intraperitoneal inoculation of ovarian cancer cells, drug treatment was administered. The mice were randomly divided into a normal saline treatment control group, BEV treatment group, pentamidine treatment group, and BEV with pentamidine treatment group. (B) Line chart of the tumor fluorescence intensity of the four groups of mice. (C) Tumor fluorescence imaging of mice after tumor formation (week 2), after killing the control group and pentamidine treatment group (week 4–6), after killing the BEV treatment group (week 8–9), and after killing the BEV with pentamidine treatment group (week 13–14). (D) Kaplan–Meier survival curves of the four groups of mice. (E) CD31 immunohistochemical staining and (F) MVD statistics of tumor tissue from the four groups of mice. (E) S100B immunohistochemical staining and S100B (G) staining intensity of tumor tissue from the four groups of mice. BEV: Bevacizumab; MVD: Micro-vessel density; *: P < 0.05; **: P < 0.01;^: P < 0.05 vs. NC; #: P < 0.05 vs. pentamidine; &: P < 0.05 vs. BEV.

Article Snippet: During phenotype experiments with HUVECs, recombinant S100B protein (MCE, HY- P70659 ) was added at concentrations of 10, 100, and 1000 mg/mL.

Techniques: In Vivo, Saline, Control, Fluorescence, Imaging, Immunohistochemical staining, Staining