The zebrafish/tumor xenograft angiogenesis assay as a tool for screening anti-angiogenic miRNAs |
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Authors: | Elena Chiavacci Milena Rizzo Letizia Pitto Francesca Patella Monica Evangelista Laura Mariani Giuseppe Rainaldi |
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Institution: | 1. Institute of Clinical Physiology, Area della Ricerca CNR, Via Moruzzi, 156124, Pisa, Italy
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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.Electronic supplementary materialThe online version of this article (doi:10.1007/s10616-014-9735-y) contains supplementary material, which is available to authorized users. |
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Keywords: | microRNAs Tumor angiogenesis Zebrafish/tumor xenografts Prostate tumor cells |
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