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Updated: Apr 27, 2026
In Vivo Imaging and Quantitation of the Host Angiogenic Response in Zebrafish Tumor Xenografts
Published on: August 14, 2019
The zebrafish/tumor xenograft angiogenesis assay as a tool for screening anti-angiogenic miRNAs
Elena Chiavacci1, Milena Rizzo1, Letizia Pitto1
1Institute of Clinical Physiology, Area della Ricerca CNR, Via Moruzzi, 156124, Pisa, Italy.
Abstract:
The zebrafish/tumor xenograft angiogenesis assay is used to approach tumor angiogenesis, a pivotal step in cancer progression and target for anti-tumor therapies. Here, we evaluated whether the assay could allow the identification of microRNAs having an anti-angiogenic potential. For that, we transfected DU-145 prostate cancer cells with four microRNAs (miR-125a, miR-320, miR-487b, miR-492) responsive to both anti- and pro-angiogenic stimuli applied to human umbilical vein endothelial cells. After transfection, DU-145 cells were injected close to the developing subintestinal vessels of transgenic Tg(Kdrl:eGFP)s843 zebrafish embryos that express green fluorescent protein under the control of Kdrl promoter. At 72 h post-fertilization, we observed that green fluorescent protein-positive neo-vessels infiltrated the graft of DU-145 transfected with miR-125a, miR-320, and miR-487b. Vice versa, neo-vessel formation and tumor cell infiltration were inhibited when DU-145 cells transfected with miR-492 were used. These results indicated that the zebrafish/tumor xenograft assay was adequate to identify microRNAs able to suppress the release of angiogenic growth factors by angiogenic tumor cells.
Insights
The zebrafish xenograft model effectively identified microRNAs targeting tumor angiogenesis. Specifically, miR-492 suppressed new blood vessel formation, highlighting its anti-angiogenic potential for cancer therapy.
Area of Science:
- Oncology
- Molecular Biology
- Developmental Biology
Background:
- Tumor angiogenesis is crucial for cancer progression and a key target for anti-cancer treatments.
- MicroRNAs (miRNAs) play significant roles in regulating angiogenesis.
- The zebrafish xenograft model offers a potential platform for identifying miRNAs with anti-angiogenic properties.
Purpose of the Study:
- To evaluate the utility of the zebrafish/tumor xenograft assay for identifying microRNAs with anti-angiogenic potential.
- To investigate the effects of specific microRNAs (miR-125a, miR-320, miR-487b, miR-492) on tumor angiogenesis in a zebrafish model.
Main Methods:
- DU-145 prostate cancer cells were transfected with four selected microRNAs.
- Transfected cells were injected into zebrafish embryos (Tg(Kdrl:eGFP)s843) near developing subintestinal vessels.
- Neovascularization and tumor cell infiltration were assessed at 72 hours post-fertilization by monitoring GFP expression.
Main Results:
- Zebrafish embryos injected with DU-145 cells transfected with miR-125a, miR-320, and miR-487b showed infiltration by GFP-positive neo-vessels.
- Conversely, DU-145 cells transfected with miR-492 significantly inhibited neo-vessel formation and tumor cell infiltration.
- This indicates differential effects of the tested microRNAs on angiogenesis.
Conclusions:
- The zebrafish/tumor xenograft assay is suitable for identifying microRNAs that can suppress angiogenic growth factor release from tumor cells.
- miR-492 demonstrates significant anti-angiogenic potential, suggesting its therapeutic relevance in prostate cancer.
- This model provides a valuable tool for preclinical screening of miRNA-based anti-angiogenic therapies.
