title: Cardiotonic steroids induce anti-angiogenic and anti-proliferative profiles in first trimester extravillous cytotrophoblast cells.
abstract: OBJECTIVE
  Preeclampsia (preE), is characterized by abnormal placental invasion and function. Marinobufagenin (MBG), a cardiotonic steroid (CTS), inhibits cytotrophoblast (CTB) cell functions that are critical for normal placental development. This study tests the hypothesis that CTSs induce anti-angiogenic and anti-proliferative effects in CTB cells.
  
  
  METHODS
  Human extravillous CTB cells of the line Sw-71, derived from first trimester chorionic villus tissue, were incubated with 0, 0.1, 1, 10, and 100 nM of each of three CTSs (MBG, cinobufatalin (CINO) and ouabain (OUB)) for 48 h. Thereafter, levels of pro-angiogenic (vascular endothelial growth factor (VEGF165), placental growth factor (PlGF)) and anti-angiogenic (soluble fms-like tyrosine kinase-1 (sFlt-1), soluble endoglin (sEng)) factors were measured in culture media using ELISA kits. Expression of three receptors (VEGF receptor 1 (VEGFR1), angiogenic angiotensin type 1 receptor (AT1) and anti-angiogenic angiotensin type 2 receptor (AT2)) were assayed using immunoblotting (western blots) in cell lysates.
  
  
  RESULTS
  sFlt-1 and sEng secretion were increased while VEGF165 and PIGF were decreased in the culture media of CTB cells treated with 1 nM or more of each CTSs (p < 0.01 for each). The AT2 receptor expression was up-regulated (p < 0.05) in CTB cells treated with 1 nM or more of MBG and CINO and with 100 nM OUB, while AT1 and VEGFR1 expressions decreased (p < 0.05) with 1 nM or more of MBG and 10 nM or more of CINO and OUB.
  
  
  CONCLUSIONS
  CTSs influence extravillous CTB cells to induce an anti-angiogenic and anti-proliferative profile.
authors.affiliation: From the Departments of Obstetrics & Gynecology, Baylor Scott & White Health and Texas A&M Health Science Center College of Medicine, Temple, TX 76508, USA.
authors.forename: J C
authors.initials: JC
authors.lastname: Ehrig
authors.affiliation: From the Departments of Obstetrics & Gynecology, Baylor Scott & White Health and Texas A&M Health Science Center College of Medicine, Temple, TX 76508, USA.
authors.forename: D
authors.initials: D
authors.lastname: Horvat
authors.affiliation: From the Departments of Obstetrics & Gynecology, Baylor Scott & White Health and Texas A&M Health Science Center College of Medicine, Temple, TX 76508, USA.
authors.forename: S R
authors.initials: SR
authors.lastname: Allen
authors.affiliation: From the Departments of Obstetrics & Gynecology, Baylor Scott & White Health and Texas A&M Health Science Center College of Medicine, Temple, TX 76508, USA.
authors.forename: R O
authors.initials: RO
authors.lastname: Jones
authors.affiliation: From the Departments of Obstetrics & Gynecology, Baylor Scott & White Health and Texas A&M Health Science Center College of Medicine, Temple, TX 76508, USA; Pediatrics, Baylor Scott & White Health and Texas A&M Health Science Center College of Medicine, Temple, TX 76508, USA; Molecular & Cellular Medicine, Baylor Scott & White Health and Texas A&M Health Science Center College of Medicine, Temple, TX 76508, USA.
authors.forename: T J
authors.initials: TJ
authors.lastname: Kuehl
authors.affiliation: From the Departments of Obstetrics & Gynecology, Baylor Scott & White Health and Texas A&M Health Science Center College of Medicine, Temple, TX 76508, USA; Pediatrics, Baylor Scott & White Health and Texas A&M Health Science Center College of Medicine, Temple, TX 76508, USA. Electronic address: MNUDDIN@sw.org.
authors.forename: M N
authors.initials: MN
authors.lastname: Uddin
mesh_terms: D002018:Bufanolides / Q000494:pharmacology; D002301:Cardiac Glycosides / Q000494:pharmacology*; D002316:Cardiotonic Agents / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010042:Ouabain / Q000494:pharmacology; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D044139:Receptor, Angiotensin, Type 2 / Q000378:metabolism; D012084:Renin-Angiotensin System; D014327:Trophoblasts / Q000187:drug effects* / Q000378:metabolism
chemical_list: D002018:Bufanolides; D002301:Cardiac Glycosides; D002316:Cardiotonic Agents; D044139:Receptor, Angiotensin, Type 2; C093896:marinobufagenin; D010042:Ouabain
keywords: Angiogenesis; Cardiac steroids; Cell proliferation; Cytotrophoblast cells