title: Rapid and cost-effective xenograft hepatocellular carcinoma model in Zebrafish for drug testing.
abstract: We developed a novel, rapid and cost-effective Zebrafish xenograft model of hepatocellular carcinoma (HCC) for drug screening in the disease. Following injection into the yolk sack of Zebrafish larvae of the human HCC cell line JHH6 stained by a vital dye, tumor mass growth was followed by fluorescence microscopy and by human Ki67 quantification. Tumor induced neo-angiogenesis was evaluated by alkaline phosphatase staining of the vessels, by using the Tg(fli1:EGFP)y1strain of Zebrafish and by the quantification of the zebrafish vascular endothelial growth factor and of its receptor. We show that it is feasible to micro-inject JHH6 in Zebrafish larvae, that injected cells can grow for different days and that this induces a marked neo-angiogenesis. Finally, we show that our model allows testing the effects of anti-HCC drugs such as Bortezomib. Compared to more complex HCC mouse models, our model is far less expensive, faster to set up and does not need immunosuppressant treatment. Finally, the model makes use of JHH6, an aggressive form of HCC cell line never tested before in Zebrafish. In conclusion, the possibility to test anti HCC/neo-angiogenesis drugs makes our JHH6 model useful to select therapeutic molecules for a highly vascularized tumor such as HCC.
authors.affiliation: Department of Medical, Surgery and Health Sciences, University of Trieste, Cattinara Hospital, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Tonon
authors.affiliation: Department of Medical, Surgery and Health Sciences, University of Trieste, Cattinara Hospital, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Zennaro
authors.affiliation: Department of Life Sciences, University of Trieste, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Dapas
authors.affiliation: Department of Medical, Surgery and Health Sciences, University of Trieste, Cattinara Hospital, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Carraro
authors.affiliation: IRCCS Galeazzi Orthopedic Institute Milan, Italy; Department of Biomedical, Surgical and Dental Sciences, University of Milan, Milan, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Mariotti
authors.affiliation: Department of Life Sciences, University of Trieste, Italy. Electronic address: ggrassi@units.it.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Grassi
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D000069286:Bortezomib / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000473:pathology*; D045744:Cell Line, Tumor; D006801:Humans; D007814:Larva; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000473:pathology*; D008845:Microinjections / Q000379:methods; D009368:Neoplasm Transplantation / Q000379:methods; D023041:Xenograft Model Antitumor Assays / Q000379:methods*; D015027:Zebrafish
chemical_list: D000970:Antineoplastic Agents; D000069286:Bortezomib
keywords: Bortezomib; Hepatocellular carcinoma; JHH6 cell line; Neo-angiogenesis; Zebrafish larvae xenograft