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Updated: Aug 6, 2026
Preparation Of Neovascular Tissues from Human Glioma Tissues for Quantitative Proteomics Analysis of Tumor Angiogenesis
Published on: March 20, 2026
Effect of bortezomib on human neuroblastoma cell growth, apoptosis, and angiogenesis
Chiara Brignole1, Danilo Marimpietri, Fabio Pastorino
1Laboratory of Oncology, G. Gaslini Children's Hospital, Largo G. Gaslini 5, 16147 Genoa, Italy.
Background:
Bortezomib is a selective and reversible inhibitor of the 26S proteasome that shows potent antitumor activity in vitro and in vivo against several human cancers of adulthood. No data are available on bortezomib activity against human pediatric neuroblastoma.
Methods:
Ten neuroblastoma cell lines and suspensions of primary neuroblastoma cells from three patients were tested for sensitivity to bortezomib. Colony formation, cell proliferation, cell cycle progression, and apoptosis were evaluated by a clonogenic assay and by measuring 3H-thymidine incorporation, bromodeoxyuridine uptake, DNA fragmentation, and phosphatidylserine exposure and propidium iodide staining, respectively. Angiogenesis was assessed by the chick embryo chorioallantoic membrane (CAM) assay. Two mouse xenograft models that mimic the growth and spread of neuroblastoma in humans were used to examine in vivo sensitivity of neuroblastoma to bortezomib. All statistical tests were two-sided.
Results:
Bortezomib inhibited proliferation and colony formation of neuroblastoma cell lines in a time- and dose-dependent manner. The mean bortezomib concentration that caused 50% inhibition of growth was 6.1 nM (95% confidence interval [CI] = 0.9 to 11.3 nM) at 72 hours. Bortezomib-treated neuroblastoma cells were arrested at G2/M and underwent apoptosis (mean percentage of apoptotic cells in four neuroblastoma cell lines treated with 20 nM bortezomib for 24 hours ranged from 20% to 35%, and caspases were activated by two- to fivefold with respect to untreated cells). Similar results were obtained for primary neuroblastoma cells exposed to bortezomib. Bortezomib inhibited angiogenesis in CAMs stimulated by conditioned medium from neuroblastoma cell lines, by neuroblastoma xenografts, and by primary neuroblastoma biopsy specimens (microvessel area: 2.9 x 10(-2) mm2, 95% CI = 1.8 x 10(-2) to 3.8 x 10(-2) mm2 in CAMs treated with biopsy specimens alone and 1.3 x 10(-2) mm2, 95% CI = 1 x 10(-2) to 1.5 x 10(-2) mm2 in CAMs treated with biopsy specimens plus bortezomib, P = .024). In both mouse models, mice treated with bortezomib lived statistically significantly longer than control mice (mean survival time in the pseudometastatic model: 74.2 versus 50.3 days, P<.001; mean survival time in the orthotopic model: 72.3 versus 50.6 days, P<.001).
Conclusions:
Bortezomib is an effective inhibitor of neuroblastoma cell growth and angiogenesis. These findings provide the rationale for further clinical investigation of bortezomib in pediatric neuroblastoma.
Insights
Bortezomib effectively inhibits pediatric neuroblastoma growth and angiogenesis in vitro and in vivo. This proteasome inhibitor demonstrates significant potential for clinical trials in treating neuroblastoma.
Area of Science:
- Oncology
- Pharmacology
- Pediatric Cancer Research
Background:
- Bortezomib is a proteasome inhibitor with established antitumor activity in adult cancers.
- Its efficacy against pediatric neuroblastoma remains largely uninvestigated.
Purpose of the Study:
- To evaluate the in vitro and in vivo activity of bortezomib against human pediatric neuroblastoma.
- To assess bortezomib's impact on neuroblastoma cell proliferation, apoptosis, and angiogenesis.
Main Methods:
- Neuroblastoma cell lines and primary cells were tested for bortezomib sensitivity.
- Assays included clonogenic assays, cell cycle analysis, apoptosis detection, and angiogenesis assessment.
- In vivo efficacy was evaluated using two mouse xenograft models.
Main Results:
- Bortezomib inhibited neuroblastoma cell proliferation and colony formation in a dose-dependent manner (IC50 = 6.1 nM).
- Cells exhibited G2/M arrest and apoptosis, with activated caspases.
- Bortezomib significantly reduced angiogenesis in CAM assays and prolonged survival in mouse models.
Conclusions:
- Bortezomib demonstrates potent anti-neuroblastoma activity by inhibiting cell growth and angiogenesis.
- These findings support clinical investigation of bortezomib for pediatric neuroblastoma treatment.
