Prolactin inhibits EGF-induced DNA synthesis in mammary epithelium via early signaling mechanisms: possible involvement of protein kinase C.
Prolactin and prolactin agonists inhibited EGF-induced DNA synthesis in mammary epithelium, whereas other pituitary hormones had no effect on EGF-induced DNA synthesis. The inhibitory effect of prolactin was seen for EGF and TGF-alpha, but not for IGF-I or cholera toxin. Autoradiography indicated that prolactin decreased the ability of EGF to induce cells to progress to S phase of the cell cycle, and time course studies indicated that the effects of prolactin were not due to an altered timing of DNA synthesis induction. Prolactin addition within 30 min of adding EGF was necessary to inhibit EGF-induced DNA synthesis. Conditioned media from prolactin-treated cells from which prolactin had been neutralized with the extracellular domain of the prolactin receptor had no effect on EGF-induced DNA synthesis, suggesting that the effect was due to prolactin, not an autocrine factor induced by prolactin. Prolactin induced a rapid association of protein kinase C with the membrane fraction of NMuMG cells, as well as increased threonine phosphorylation of the EGF receptor. Protein kinase C inhibitors eliminated most of the inhibitory effect of prolactin on EGF-induced DNA synthesis. The protein kinase C inhibitor Calphostin C restored high-affinity EGF binding in prolactin-treated cells and reversed the inhibitory effect of prolactin on EGF-induced EGF receptor tyrosine phosphorylation.
Fenton
S E
SE
Endocrinology-Reproductive Physiology Program, University of Wisconsin, Madison 53706, USA.
Sheffield
L G
LG
D000818:Animals; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000037:antagonists & inhibitors / Q000502:physiology*; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D066246:ErbB Receptors / Q000187:drug effects / Q000378:metabolism; D007211:Indoles / Q000494:pharmacology; D007457:Iodine Radioisotopes; D008301:Maleimides / Q000494:pharmacology; D008321:Mammary Glands, Animal / Q000166:cytology*; D051379:Mice; D009281:Naphthalenes / Q000494:pharmacology; D010766:Phosphorylation; D011388:Prolactin / Q000378:metabolism / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012680:Sensitivity and Specificity; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology; D013912:Threonine / Q000378:metabolism
D004791:Enzyme Inhibitors; D007211:Indoles; D007457:Iodine Radioisotopes; D008301:Maleimides; D009281:Naphthalenes; D013912:Threonine; D004815:Epidermal Growth Factor; D011388:Prolactin; D004247:DNA; D066246:ErbB Receptors; D011493:Protein Kinase C; C058819:calphostin C; C088060:bisindolylmaleimide