Generation of liver-specific TGF-α and c-Myc-overexpressing fibroblasts for future creation of a liver cancer porcine model

Mol Med Rep. 2014 Jul;10(1):329-35. doi: 10.3892/mmr.2014.2217. Epub 2014 May 7.

Abstract

Liver cancer is one of the most serious life-threatening diseases in the world. Although the rodent model of hepatocellar carcinoma (HCC) is commonly used, it is limited when considering preclinical applications, including transarterial chemoembolization. The pig is a more appropriate model for applying preclinical procedures as it has similar anatomical and physiological characteristics to humans. In the current study, transgenic fibroblasts were generated that overexpressed two proto-oncogenes specifically in hepatocytes. Porcine TGF-α and c-myc genes were isolated and these were linked with the porcine albumin promoter, which has exhibited selective activity in liver cells. Targeting vectors were introduced into the porcine fibroblasts using a liposome-mediated delivery system and the transgenic cell line was screened with 3 weeks of G-418 treatment. Selected vector‑positive colonies were further confirmed with polymerase chain reaction-based genotyping. Thus, the transgenic cell lines created in the current study should induce liver cancer in pig models following somatic cell nuclear transfer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Albumins / genetics
  • Animals
  • Cell Line
  • Disease Models, Animal
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • HEK293 Cells
  • Hep G2 Cells
  • Hepatocytes / cytology
  • Hepatocytes / metabolism*
  • Humans
  • Liver Neoplasms / metabolism
  • Liver Neoplasms / pathology
  • Promoter Regions, Genetic
  • Proto-Oncogene Proteins c-myc / genetics
  • Proto-Oncogene Proteins c-myc / metabolism*
  • Swine
  • Transfection
  • Transforming Growth Factor alpha / genetics
  • Transforming Growth Factor alpha / metabolism*

Substances

  • Albumins
  • Proto-Oncogene Proteins c-myc
  • Transforming Growth Factor alpha