G protein-coupled receptor agonist-stimulated expression of ATF3/LRF-1 and c-myc and comitogenic effects in hepatocytes do not require EGF receptor transactivation

J Cell Physiol. 2004 Dec;201(3):349-58. doi: 10.1002/jcp.20075.

Abstract

Several agonists acting on G protein-coupled receptors (GPCR) enhance the mitogenic effect of epidermal growth factor (EGF) in rat hepatocytes, through mechanisms that have only partially been clarified. Results in various cells have led to the idea that a major mechanism for GPCR-mediated stimulation of cell growth is transactivation of receptor tyrosine kinases, particularly the EGF receptor (EGFR), leading to rapid phosphorylation of the EGFR and activation of downstream signaling pathways. In the present study cultured rat hepatocytes were exposed to various GPCR agonists, including vasopressin, angiotensin II (Ang.II), norepinephrine, or prostaglandin F(2 alpha) (PGF(2 alpha)). None of these agents increased the phosphorylation of the EGFR or the docking protein Shc. Furthermore, we examined the effect of the GPCR agonists on the expression of two early response genes believed to be involved in growth activation. The GPCR agonists increased the mRNA expression of c-myc, and also of activating transcription factor 3 (ATF3)/liver regeneration factor-1 (LRF-1), which is a novel finding. Finally, the selective EGFR inhibitor AG1478 did not suppress the activation of extracellular signal-regulated kinase 1/2 (ERK1/2) or the induction of c-myc or ATF3/LRF-1 by the GPCR agonists, and did not prevent the comitogenic effects induced by these agents, while it blocked the effect of EGF on these responses. The results suggest that GPCR agonists induce expression of ATF3/LRF-1 and c-myc and exert comitogenic effects through mechanisms that do not require EGFR transactivation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activating Transcription Factor 3
  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Cells, Cultured
  • DNA / metabolism
  • DNA-Binding Proteins / genetics*
  • Enzyme Activation / drug effects
  • Enzyme Inhibitors / pharmacology
  • ErbB Receptors / agonists
  • ErbB Receptors / metabolism*
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Focal Adhesion Kinase 2
  • Liver / cytology
  • Liver / drug effects*
  • Liver / metabolism
  • Male
  • Mitogens / pharmacology*
  • Mitosis / drug effects*
  • Phosphorylation / drug effects
  • Protein-Tyrosine Kinases / metabolism
  • Proto-Oncogene Proteins c-myc / genetics*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Wistar
  • Receptors, G-Protein-Coupled / agonists*
  • Shc Signaling Adaptor Proteins
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Transcriptional Activation / drug effects

Substances

  • Activating Transcription Factor 3
  • Adaptor Proteins, Signal Transducing
  • DNA-Binding Proteins
  • Enzyme Inhibitors
  • Mitogens
  • Proto-Oncogene Proteins c-myc
  • RNA, Messenger
  • Receptors, G-Protein-Coupled
  • Shc Signaling Adaptor Proteins
  • Shc1 protein, rat
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • liver regeneration factor 1
  • DNA
  • ErbB Receptors
  • Protein-Tyrosine Kinases
  • Focal Adhesion Kinase 2
  • Ptk2b protein, rat
  • Extracellular Signal-Regulated MAP Kinases