Retroviral gene transfer of signaling molecules into murine fetal hepatocytes defines distinct roles for the STAT3 and ras pathways during hepatic development

Hepatology. 2000 Dec;32(6):1370-6. doi: 10.1053/jhep.2000.19815.

Abstract

We recently demonstrated that oncostatin M (OSM) in the presence of glucocorticoid promotes development of fetal hepatic cells in a primary culture system. Our results also suggested that OSM transduces differentiation signals through gp130, a common subunit of the interleukin (IL)-6 family cytokine receptors. However, an essential downstream pathway required for hepatic development remains unknown. To address this issue, we expressed signal molecules by a retroviral expression vector in primary fetal hepatic cells and investigated a signaling pathway essential for OSM-mediated hepatic development. Expression of a dominant-negative form of STAT3 (DeltaSTAT3), but not DeltaSTAT5, suppressed differentiated phenotypes of hepatocytes induced by OSM. On the other hand, dominant-negative forms of Ras (RasN17) and SHP-2(C463A) rather augmented the expression of hepatic differentiation markers, suggesting that the Ras pathway negatively regulates hepatic development. Consistently, expression of a constitutively activated form of Ras (RasV12) inhibited cellular responses to OSM. Our results indicate that STAT3 is an essential signaling component for OSM-induced hepatic development, while activation of Ras appears to negatively regulate this process. Thus, retrovirus-mediated gene transfer is an effective means to analyze function of a gene in primary fetal hepatic culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers
  • Cell Differentiation / physiology
  • Chromones / pharmacology
  • DNA-Binding Proteins / physiology*
  • Embryonic and Fetal Development
  • Enzyme Inhibitors / pharmacology
  • Fetus / physiology
  • Gene Expression
  • Gene Transfer Techniques*
  • Genetic Vectors
  • Hepatocytes / physiology*
  • Liver / cytology
  • Liver / embryology*
  • Mice
  • Morpholines / pharmacology
  • Oncostatin M
  • Peptides / physiology
  • Phosphoinositide-3 Kinase Inhibitors
  • Retroviridae / genetics*
  • STAT3 Transcription Factor
  • Signal Transduction / physiology*
  • Trans-Activators / physiology*
  • Transgenes / genetics
  • ras Proteins / physiology*

Substances

  • Biomarkers
  • Chromones
  • DNA-Binding Proteins
  • Enzyme Inhibitors
  • Morpholines
  • Osm protein, mouse
  • Peptides
  • Phosphoinositide-3 Kinase Inhibitors
  • STAT3 Transcription Factor
  • Stat3 protein, mouse
  • Trans-Activators
  • Oncostatin M
  • 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
  • ras Proteins