title: Locostatin, a disrupter of Raf kinase inhibitor protein, inhibits extracellular matrix production, proliferation, and migration in human uterine leiomyoma and myometrial cells.
abstract: OBJECTIVE
  To investigate the presence of Raf kinase inhibitor protein (RKIP) in human myometrium and leiomyoma as well as to determine the effect of locostatin (RKIP inhibitor) on extracellular matrix (ECM) production, proliferation, and migration in human myometrial and leiomyoma cells.
  
  
  METHODS
  Laboratory study.
  
  
  METHODS
  Human myometrium and leiomyoma.
  
  
  METHODS
  Thirty premenopausal women who were admitted to the hospital for myomectomy or hysterectomy.
  
  
  METHODS
  Myometrial and leiomyoma tissues were used to investigate the localization and the expression level of RKIP through immunohistochemistry and Western blotting. Myometrial and leiomyoma cells were treated with locostatin (10 μM) to measure ECM expression by real-time polymerase chain reaction, GSK3β expression by Western blotting, cell migration by wound-healing assay, and cell proliferation by MTT assay and immunocytochemistry.
  
  
  METHODS
  The expression of RKIP in human myometrial and leiomyoma tissue; ECM components and GSK3β expression, migration, and proliferation in myometrial and leiomyoma cells.
  
  
  RESULTS
  RKIP is expressed in human myometrial and leiomyoma tissue. Locostatin treatment resulted in the activation of the mitogen-activated protein kinase (MAPK) signal pathway (ERK phosphorylation), providing a powerful validation of our targeting protocol. Further, RKIP inhibition by locostatin reduces ECM components. Moreover, the inhibition of RKIP by locostatin impaired cell proliferation and migration in both leiomyoma and myometrial cells. Finally, locostatin treatment reduced GSK3β expression. Therefore, even if the activation of MAPK pathway should increase proliferation and migration, the destabilization of GSK3β leads to the reduction of proliferation and migration of myometrial and leiomyoma cells.
  
  
  CONCLUSIONS
  Our results indicate that RKIP may be involved in leiomyoma pathophysiology.
authors: Department of Experimental and Clinical Medicine, Faculty of Medicine, Università Politecnica delle Marche, Ancona, Italy.
authors: Milijana
authors: M
authors: Janjusevic
authors: Department of Experimental and Clinical Medicine, Faculty of Medicine, Università Politecnica delle Marche, Ancona, Italy.
authors: Stefania
authors: S
authors: Greco
authors: Department of Experimental and Clinical Medicine, Faculty of Medicine, Università Politecnica delle Marche, Ancona, Italy; Biotechnology and Microbiology Laboratory, Department of Botany, University of Rajshahi, Rajshahi, Bangladesh.
authors: Md Soriful
authors: MS
authors: Islam
authors: Department of Clinical and Molecular Sciences, Università Politecnica delle Marche, Ancona, Italy.
authors: Clara
authors: C
authors: Castellucci
authors: Department of Clinical Science, Università Politecnica delle Marche, Ancona, Italy.
authors: Andrea
authors: A
authors: Ciavattini
authors: Department of Medical Biotechnology, University of Siena, Siena, Italy.
authors: Paolo
authors: P
authors: Toti
authors: Department of Molecular and Developmental Medicine, Obstetrics, and Gynecology, University of Siena, Siena, Italy.
authors: Felice
authors: F
authors: Petraglia
authors: Department of Experimental and Clinical Medicine, Faculty of Medicine, Università Politecnica delle Marche, Ancona, Italy; Department of Information Engineering, Università Politecnica delle Marche, Ancona, Italy. Electronic address: p.ciarmela@univpm.it.
authors: Pasquapina
authors: P
authors: Ciarmela
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D005109:Extracellular Matrix / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D000071679:Glycogen Synthase Kinase 3 beta / Q000378:metabolism; D006801:Humans; D007889:Leiomyoma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D009215:Myometrium / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D023303:Oxazolidinones / Q000494:pharmacology*; D049488:Phosphatidylethanolamine Binding Protein / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: D000970:Antineoplastic Agents; D023303:Oxazolidinones; C511482:PEBP1 protein, human; D049488:Phosphatidylethanolamine Binding Protein; C529490:locostatin; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: RKIP; Uterine leiomyoma; cell migration; extracellular matrix; locostatin