Identification and Characterization of Metastatic Factors by Gene Transfer into the Novel RIP-Tag; RIP-tva Murine Model

J Vis Exp. 2017 Oct 16:(128):55890. doi: 10.3791/55890.

Abstract

Metastatic cancer accounts for 90% of deaths in patients with solid tumors. There is an urgent need to better understand the drivers of cancer metastasis and to identify novel therapeutic targets. To investigate molecular events that drive the progression from primary cancer to metastasis, we have developed a bitransgenic mouse model, RIP-Tag; RIP-tva. In this mouse model, the rat insulin promoter (RIP) drives the expression of the SV40 T antigen (Tag) and the receptor for subgroup A avian leukosis virus (tva) in pancreatic β cells. The mice develop pancreatic neuroendocrine tumors with 100% penetrance through well-defined stages that are similar to human tumorigenesis, with stages including hyperplasia, angiogenesis, adenoma, and invasive carcinoma. Because RIP-Tag; RIP-tva mice do not develop metastatic disease, genetic alterations that promote metastasis can be identified easily. Somatic gene transfer into tva-expressing, proliferating pancreatic β premalignant lesions is achieved through intracardiac injection of avian retroviruses harboring the desired genetic alteration. A titer of >1 x 108 infectious units per ml is considered appropriate for in vivo infection. In addition, avian retroviruses can infect cell lines derived from tumors in RIP-Tag; RIP-tva mice with high efficiency. The cell lines can also be used to characterize the metastatic factors. Here we demonstrate how to utilize this mouse model and cell lines to assess the functions of candidate genes in tumor metastasis.

Publication types

  • Video-Audio Media

MeSH terms

  • Animals
  • Antigens, Polyomavirus Transforming / biosynthesis
  • Antigens, Polyomavirus Transforming / genetics
  • Avian Leukosis Virus / genetics*
  • Disease Models, Animal
  • Gene Transfer Techniques*
  • Genetic Vectors / genetics
  • Insulin / genetics
  • Insulin-Secreting Cells / pathology
  • Insulin-Secreting Cells / virology
  • Mice
  • Mice, Transgenic
  • Neoplasm Metastasis
  • Neuroendocrine Tumors / genetics*
  • Neuroendocrine Tumors / pathology*
  • Pancreatic Neoplasms / genetics*
  • Pancreatic Neoplasms / pathology*
  • Pancreatic Neoplasms / virology
  • Precancerous Conditions / genetics
  • Precancerous Conditions / pathology
  • Promoter Regions, Genetic

Substances

  • Antigens, Polyomavirus Transforming
  • Insulin