Cytotechnology

, Volume 67, Issue 6, pp 969–975

The zebrafish/tumor xenograft angiogenesis assay as a tool for screening anti-angiogenic miRNAs

  • Elena Chiavacci
  • Milena Rizzo
  • Letizia Pitto
  • Francesca Patella
  • Monica Evangelista
  • Laura Mariani
  • Giuseppe Rainaldi
Original Research

DOI: 10.1007/s10616-014-9735-y

Cite this article as:
Chiavacci, E., Rizzo, M., Pitto, L. et al. Cytotechnology (2015) 67: 969. doi:10.1007/s10616-014-9735-y

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.

Keywords

microRNAs Tumor angiogenesis Zebrafish/tumor xenografts Prostate tumor cells 

Supplementary material

10616_2014_9735_MOESM1_ESM.doc (58 kb)
Supplementary material 1 (DOC 59 kb)

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Elena Chiavacci
    • 1
  • Milena Rizzo
    • 1
  • Letizia Pitto
    • 1
  • Francesca Patella
    • 1
  • Monica Evangelista
    • 1
  • Laura Mariani
    • 1
  • Giuseppe Rainaldi
    • 1
  1. 1.Institute of Clinical PhysiologyArea della Ricerca CNRPisaItaly

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