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Dis Model Mech. 2014 Jul;7(7):883-94. doi: 10.1242/dmm.014969. Epub 2014 May 30.

Zebrafish reporter lines reveal in vivo signaling pathway activities involved in pancreatic cancer.

Author information

1
Department of Biology, University of Padua, 35131 Padua, Italy.
2
Department of Molecular Medicine, University of Padua, 35131 Padua, Italy.
3
Department of Woman and Child Health, University of Padua, 35131 Padua, Italy.
4
Department of Surgery and The McKusick-Nathans Institute of Genetic Medicine Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
5
Department of Biology, University of Padua, 35131 Padua, Italy. francesco.argenton@unipd.it.

Abstract

Pancreatic adenocarcinoma, one of the worst malignancies of the exocrine pancreas, is a solid tumor with increasing incidence and mortality in industrialized countries. This condition is usually driven by oncogenic KRAS point mutations and evolves into a highly aggressive metastatic carcinoma due to secondary gene mutations and unbalanced expression of genes involved in the specific signaling pathways. To examine in vivo the effects of KRAS(G12D) during pancreatic cancer progression and time correlation with cancer signaling pathway activities, we have generated a zebrafish model of pancreatic adenocarcinoma in which eGFP-KRAS(G12D) expression was specifically driven to the pancreatic tissue by using the GAL4/UAS conditional expression system. Outcrossing the inducible oncogenic KRAS(G12D) line with transgenic zebrafish reporters, harboring specific signaling responsive elements of transcriptional effectors, we were able to follow TGFβ, Notch, Bmp and Shh activities during tumor development. Zebrafish transgenic lines expressing eGFP-KRAS(G12D) showed normal exocrine pancreas development until 3 weeks post fertilization (wpf). From 4 to 24 wpf we observed several degrees of acinar lesions, characterized by an increase in mesenchymal cells and mixed acinar/ductal features, followed by progressive bowel and liver infiltrations and, finally, highly aggressive carcinoma. Moreover, live imaging analysis of the exocrine pancreatic tissue revealed an increasing number of KRAS-positive cells and progressive activation of TGFβ and Notch pathways. Increase in TGFβ, following KRAS(G12D) activation, was confirmed in a concomitant model of medulloblastoma (MDB). Notch and Shh signaling activities during tumor onset were different between MDB and pancreatic adenocarcinoma, indicating a tissue-specific regulation of cell signaling pathways. Moreover, our results show that a living model of pancreatic adenocarcinoma joined with cell signaling reporters is a suitable tool for describing in vivo the signaling cascades and molecular mechanisms involved in tumor development and a potential platform to screen for novel oncostatic drugs.

KEYWORDS:

KRAS; Medulloblastoma; Notch; Pancreatic adenocarcinoma; Reporters; Shh; TGFβ; Zebrafish

PMID:
24878567
PMCID:
PMC4073277
DOI:
10.1242/dmm.014969
[Indexed for MEDLINE]
Free PMC Article

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