title: Both estrogen and raloxifene cause G1 arrest of vascular smooth muscle cells.
abstract: The proliferation of vascular smooth muscle cells (VSMC) is a crucial pathophysiological process in the development of atherosclerosis. Although estrogen is known to inhibit the proliferation of VSMC, the mechanism responsible for this effect remains to be elucidated. In addition, the effect of raloxifene on VSMC remains unknown. We have shown here that 17beta-estradiol (E(2)) and raloxifene significantly inhibited the platelet-derived growth factor (PDGF)-stimulated proliferation of cultured human VSMC. Flow cytometry demonstrated that PDGF-stimulated S-phase progression of the cell cycle in VSMC was also suppressed by E(2) or raloxifene. We found that PDGF-induced phosphorylation of retinoblastoma protein (pRb), whose hyperphosphorylation is a hallmark of the G1-S transition in the cell cycle, was significantly inhibited by E(2) and raloxifene. These effects were associated with a decrease in cyclin D1 expression, without a change in cyclin-dependent kinase 4 or cyclin-dependent kinase inhibitor, p27(kip1) expression. ICI 182,780 abolished the inhibitory effects of E(2) and raloxifene on PDGF-induced pRb phosphorylation. Next, we examined which estrogen receptor (ER) is necessary for these effects of E(2) and raloxifene. Since VSMC express both ERalpha and ERbeta, A10, a rat aortic smooth muscle cell line that expresses ERbeta but not ERalpha, was used. The dose-dependent stimulation of A10 cell proliferation by PDGF was not inhibited by E(2) or raloxifene in contrast to the results obtained in VSMC. Moreover, E(2) and raloxifene significantly inhibited the PDGF-induced cyclin D1 promoter activity in A10 cells transfected with cDNA for ERalpha but not in the parental cells. These results suggested that E(2) and raloxifene exert an antiproliferative effect in VSMC treated with PDGF, at least in part through inhibition of pRb phosphorylation, and that the inhibitory effects of E(2) and raloxifene may be mainly mediated by ERalpha.
authors.affiliation: Department of Obstetrics and Gynecology, Yamagata University School of Medicine, 2-2-2 Iidanishi, Yamagata, Yamagata 990-9585, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Takahashi
authors.forename: M
authors.initials: M
authors.lastname: Ohmichi
authors.forename: M
authors.initials: M
authors.lastname: Yoshida
authors.forename: K
authors.initials: K
authors.lastname: Hisamoto
authors.forename: S
authors.initials: S
authors.lastname: Mabuchi
authors.forename: E
authors.initials: E
authors.lastname: Arimoto-Ishida
authors.forename: A
authors.initials: A
authors.lastname: Mori
authors.forename: S
authors.initials: S
authors.lastname: Tsutsumi
authors.forename: K
authors.initials: K
authors.lastname: Tasaka
authors.forename: Y
authors.initials: Y
authors.lastname: Murata
authors.forename: H
authors.initials: H
authors.lastname: Kurachi
mesh_terms: D000818:Animals; D001011:Aorta; D015153:Blotting, Western / Q000379:methods; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002478:Cells, Cultured; D019938:Cyclin D1 / Q000378:metabolism; D003864:Depression, Chemical; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology*; D047628:Estrogen Receptor alpha; D005434:Flow Cytometry; D000077267:Fulvestrant; D016193:G1 Phase; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D020849:Raloxifene Hydrochloride / Q000494:pharmacology*; D051381:Rats; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020845:Selective Estrogen Receptor Modulators / Q000494:pharmacology*; D014162:Transfection / Q000379:methods
chemical_list: D047628:Estrogen Receptor alpha; D010982:Platelet-Derived Growth Factor; D011960:Receptors, Estrogen; D016160:Retinoblastoma Protein; D020845:Selective Estrogen Receptor Modulators; D019938:Cyclin D1; D000077267:Fulvestrant; D020849:Raloxifene Hydrochloride; D004958:Estradiol