Dissociation between insulin-mediated signaling pathways and biological effects in placental cells: role of protein kinase B and MAPK phosphorylation.
Beyond the presence of insulin receptors, little is known of the mechanisms underlying the biological effects of insulin in the placenta. We show that phosphorylation of MAPK and protein kinase B were enhanced 286 +/- 23% and 393 +/- 17% upon insulin stimulation of JAr placental cells. MAPK activation was prevented by pretreatment with PD98059 but was unaffected by wortmannin. Insulin stimulation of protein kinase B phosphorylation was abolished by pretreatment with wortmannin, suggesting that it is dependent on phosphatidylinositol 3- kinase activation. Despite protein kinase B phosphorylation, GLUT4 translocation, glucose uptake, and glycogen synthesis were not stimulated by insulin. By contrast, glycogen synthesis was stimulated 20-fold in cells incubated with 11 mM glucose. Mitogenesis assessed by incorporation of [(3)H]thymidine into DNA was enhanced 1.9-fold in response to insulin. Stimulation of DNA synthesis was inhibited by pretreatment with PD98059 but was insensitive to wortmannin. These results indicate that stimulation of mitogenesis is one major biological effect of insulin in placenta cells that implicates the MAPK signaling pathway. Phosphatidylinositol 3-kinase- dependent protein kinase B activation is not sufficient to stimulate glucose transport and glycogen synthesis, highlighting the placenta as a nonclassic target of insulin for the regulation of glucose metabolism.
Boileau
P
P
Centre National de la Recherche Scientifique-Unité Propre de Recherche 1524, 92190 Meudon, France.
Caüzac
M
M
Pereira
M A
MA
Girard
J
J
Hauguel-De Mouzon
S
S
D006003:Glycogen / Q000096:biosynthesis; D006801:Humans; D007328:Insulin / Q000494:pharmacology / Q000502:physiology*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D008934:Mitogens / Q000494:pharmacology; D009004:Monosaccharide Transport Proteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010920:Placenta / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000378:metabolism / Q000502:physiology*; D051057:Proto-Oncogene Proteins c-akt; D011972:Receptor, Insulin / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D014407:Tumor Cells, Cultured
D007328:Insulin; D008934:Mitogens; D009004:Monosaccharide Transport Proteins; D011518:Proto-Oncogene Proteins; D006003:Glycogen; D011972:Receptor, Insulin; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D020928:Mitogen-Activated Protein Kinases