title: Role of activin-A and myostatin and their signaling pathway in human myometrial and leiomyoma cell function.
abstract: BACKGROUND
  Uterine leiomyomas are highly prevalent benign tumors of premenopausal women and the most common indication for hysterectomy. However, the exact etiology of this tumor is not fully understood.
  
  
  OBJECTIVE
  The objective of the study was to evaluate the role of activin-A and myostatin and their signaling pathways in human myometrial and leiomyoma cells.
  
  
  METHODS
  This was a laboratory study.
  
  
  METHODS
  Myometrial and leiomyoma cells (primary and cell lines) were cultured in vitro.
  
  
  METHODS
  The study included premenopausal women who were admitted to the hospital for myomectomy or hysterectomy.
  
  
  METHODS
  Primary myometrial and leiomyoma cells and/or cell lines were treated with activin-A (4 nM) and myostatin (4 nM) for different days of interval (to measure proliferation rate) or 30 minutes (to measure signaling molecules) or 48 hours to measure proliferating markers, extracellular matrix mRNA, and/or protein expression by real-time PCR, Western blot, and/or immunocytochemistry.
  
  
  RESULTS
  We found that activin-A and myostatin significantly reduce cell proliferation in primary myometrial cells but not in leiomyoma cells as measured by a CyQUANT cell proliferation assay kit. Reduced expression of proliferating cell nuclear antigen and Ki-67 were also observed in myometrial cells in response to activin-A and myostatin treatment. Activin-A also significantly increased mRNA expression of fibronectin, collagen1A1, and versican in primary leiomyoma cells. Finally, we found that activin-A and myostatin activate Smad-2/3 signaling but do not affect ERK or p38 signaling in both myometrial and leiomyoma cells.
  
  
  CONCLUSIONS
  This study results suggest that activin-A and myostatin can exert antiproliferative and/or fibrotic effects on these cell types via Smad-2/3 signaling.
authors.affiliation: Department of Experimental and Clinical Medicine (M.S.I., O.P., M.J., M.C., P.C.), Faculty of Medicine, Polytechnic University of Marche, 60126 Ancona, Italy; Biotechnology and Microbiology Laboratory (M.S.I.), Department of Botany, University of Rajshahi, Rajshahi 6205, Bangladesh; Department of Obstetrics and Gynecology (W.H.C.), Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814; Clayton Foundation Laboratories for Peptide Biology (P.C.G.), The Salk Institute for Biological Studies, La Jolla, California 92037; Departments of Clinical Science (S.R.G., A.C., A.L.T.) and Information Engineering (P.C.), Polytechnic University of Marche, 60131 Ancona, Italy; Obstetrics and Gynecology Unit (P.L.), Profili Hospital, Fabriano, 60044 Italy; Department of Molecular and Developmental Medicine, Obstetrics, and Gynecology (F.P.), University of Siena, 53100 Siena, Italy.
authors.forename: Md Soriful
authors.initials: MS
authors.lastname: Islam
authors.forename: William H
authors.initials: WH
authors.lastname: Catherino
authors.forename: Olga
authors.initials: O
authors.lastname: Protic
authors.forename: Milijana
authors.initials: M
authors.lastname: Janjusevic
authors.forename: Peter Clarke
authors.initials: PC
authors.lastname: Gray
authors.forename: Stefano Raffaele
authors.initials: SR
authors.lastname: Giannubilo
authors.forename: Andrea
authors.initials: A
authors.lastname: Ciavattini
authors.forename: Pasquale
authors.initials: P
authors.lastname: Lamanna
authors.forename: Andrea Luigi
authors.initials: AL
authors.lastname: Tranquilli
authors.forename: Felice
authors.initials: F
authors.lastname: Petraglia
authors.forename: Mario
authors.initials: M
authors.lastname: Castellucci
authors.forename: Pasquapina
authors.initials: P
authors.lastname: Ciarmela
mesh_terms: D028341:Activins / Q000494:pharmacology*; D000328:Adult; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D024042:Collagen Type I / Q000378:metabolism; D005260:Female; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D007889:Leiomyoma / Q000378:metabolism* / Q000473:pathology; D008875:Middle Aged; D009215:Myometrium / Q000378:metabolism* / Q000473:pathology; D055435:Myostatin / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000378:metabolism* / Q000473:pathology
chemical_list: D024042:Collagen Type I; D005353:Fibronectins; D055435:Myostatin; D051899:Smad2 Protein; D051900:Smad3 Protein; C422405:activin A; D028341:Activins