17-beta estradiol elicits an autocrine leiomyoma cell proliferation: evidence for a stimulation of protein kinase-dependent pathway.
The mechanism by which estradiol (E2) acts on cell proliferation is still unclear. In this paper, we report the results of a series of experiments in an attempt to elucidate the effector pathway(s) involved in coupling the E2 receptors binding to cellular growth response in leiomyoma cells (LSMC). Under conditions of E2-dependent growth, E2 treatment of LSMC triggers rapid and transient activation of the MAP-kinase pathway. Interestingly, we demonstrate that the early downstream signal transduction events determined by E2-stimulation in quiescent LSMC, including the rapid protein tyrosine phosphorylation of a subset of intracellular proteins, such GAP, PI-3-K, and PLCgamma, and the concomitant activation of ancillary protein kinases, are related to E2-induced PDGF secretion. Moreover, we identify the PDGF, alone or in association with other growth factors, as the main growth factor involved in the proliferation response of LSMC to E2 stimulation. The addition of neutralizing antibodies anti-PDGF was able to inhibit the mitogenic activity present in LSMC conditioned media samples. On the other hand, E2 did not affect the constitutive expression as well as the ligand affinity of PDGF receptors on LSMC plasmamembrane. Cell treatment with the antiestrogen ICI 182780 correlate both with a perturbation of E2-induced transductional circuit and with the disappearance of the mitogenic factor, PDGF, in LSMC conditioned media; the latter therefore, represents the main autocrine mediator of cell growth modulation, upregulated by E2 and down-regulated by antiestrogenic compound. Our experiments suggest that growth factor secretion is an initial and integral part of the signaling events mediated by the estradiol receptors, not related, at least in part, to E2 transcriptional modulation.
Barbarisi
A
A
Institute for Clinical Surgery, 2nd University of Naples, School of Medicine, Naples, Italy.
Petillo
O
O
Di Lieto
A
A
Melone
M A
MA
Margarucci
S
S
Cannas
M
M
Peluso
G
G
D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival; D017077:Culture Media, Conditioned; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology*; D020847:Estrogen Receptor Modulators / Q000494:pharmacology; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D000077267:Fulvestrant; D006801:Humans; D007700:Kinetics; D007889:Leiomyoma / Q000473:pathology* / Q000503:physiopathology; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D019000:Phosphotyrosine / Q000032:analysis; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology / Q000502:physiology; D019908:Proto-Oncogene Proteins c-raf / Q000378:metabolism; D011959:Receptors, Estradiol / Q000502:physiology; D017479:Receptors, Platelet-Derived Growth Factor / Q000502:physiology; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000473:pathology* / Q000503:physiopathology
D017077:Culture Media, Conditioned; D020847:Estrogen Receptor Modulators; D010982:Platelet-Derived Growth Factor; D011959:Receptors, Estradiol; D016222:Fibroblast Growth Factor 2; D019000:Phosphotyrosine; D000077267:Fulvestrant; D004958:Estradiol; D017479:Receptors, Platelet-Derived Growth Factor; D019908:Proto-Oncogene Proteins c-raf; D020928:Mitogen-Activated Protein Kinases