title: Altered expression of G1/S phase cell cycle regulators in placental mesenchymal stromal cells derived from preeclamptic pregnancies with fetal-placental compromise.
abstract: Herein, we evaluated whether Placental Mesenchymal Stromal Cells (PDMSCs) derived from normal and Preeclamptic (PE) placentae presented differences in the expression of G1/S-phase regulators p16INK4A, p18INK4C, CDK4 and CDK6. Finally, we investigated normal and PE-PDMSCs paracrine effects on JunB, Cyclin D1, p16INK4A, p18INK4C, CDK4 and CDK6 expressions in physiological term villous explants. PDMSCs were isolated from physiological (n = 20) and PE (n = 24) placentae. Passage three normal and PE-PDMSC and conditioned media (CM) were collected after 48h. Physiological villous explants (n = 60) were treated for 72h with normal or PE-PDMSCs CM. Explants viability was assessed by Lactate Dehydrogenase Cytotoxicity assay. Cyclin D1 localization was evaluated by Immuofluorescence (IF) while JunB, Cyclin-D1 p16INK4A, p18INK4C, CDK4 and CDK6 levels were assessed by Real Time PCR and Western Blot assay. We reported significantly increased p16INK4Aand p18INK4Cexpression in PE- relative to normal PDMSCs while no differences in CDK4 and CDK6 levels were detected. Explants viability was not affected by normal or PE-PDMSCs CM. Normal PDMSCs CM increased JunB, p16INK4and p18INK4Cand decreased Cyclin-D1 in placental tissues. In contrast, PE-PDMSCs CM induced JunB downregulation and Cyclin D1 increase in placental explants. Cyclin D1 IF staining showed that CM treatment targeted mainly the syncytiotrophoblast. We showed Cyclin D1-p16INK4A/p18INK4C altered pathway in PE-PDMSCs demonstrating an aberrant G1/S phase transition in these pathological cells. The abnormal Cyclin D1-p16INK4A/p18INK4C expression in explants conditioned by PE-PDMSCs media suggest a key contribution of mesenchymal cells to the altered trophoblast cell cycle regulation typical of PE pregnancies with fetal-placental compromise.
authors.affiliation: a Department of Surgical Sciences , University of Turin , Turin , Italy.
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Nuzzo
authors.affiliation: a Department of Surgical Sciences , University of Turin , Turin , Italy.
authors.forename: Domenica
authors.initials: D
authors.lastname: Giuffrida
authors.affiliation: b Città della Salute & della Scienza - O.I.R.M. S.Anna Hospital , Turin , Italy.
authors.forename: Bianca
authors.initials: B
authors.lastname: Masturzo
authors.affiliation: c Neurosciences Institute Cavalieri Ottolenghi (NICO), Department of Neurosciences , University of Turin, San Luigi Hospital , Orbassano , Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Mele
authors.affiliation: a Department of Surgical Sciences , University of Turin , Turin , Italy.|b Città della Salute & della Scienza - O.I.R.M. S.Anna Hospital , Turin , Italy.
authors.forename: Ettore
authors.initials: E
authors.lastname: Piccoli
authors.affiliation: c Neurosciences Institute Cavalieri Ottolenghi (NICO), Department of Neurosciences , University of Turin, San Luigi Hospital , Orbassano , Italy.
authors.forename: Carola
authors.initials: C
authors.lastname: Eva
authors.affiliation: a Department of Surgical Sciences , University of Turin , Turin , Italy.|b Città della Salute & della Scienza - O.I.R.M. S.Anna Hospital , Turin , Italy.
authors.forename: Tullia
authors.initials: T
authors.lastname: Todros
authors.affiliation: a Department of Surgical Sciences , University of Turin , Turin , Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Rolfo
mesh_terms: D000328:Adult; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000494:pharmacology; D005260:Female; D005333:Fetus / Q000473:pathology*; D016193:G1 Phase / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D010920:Placenta / Q000187:drug effects / Q000473:pathology*; D011225:Pre-Eclampsia / Q000235:genetics / Q000473:pathology*; D011247:Pregnancy; D016196:S Phase / Q000187:drug effects
chemical_list: D018797:Cell Cycle Proteins; D017077:Culture Media, Conditioned
keywords: cell cycle; mesenchymal stromal cells; placenta; preeclampsia; pregnancy