Liarozole inhibits transforming growth factor-β3--mediated extracellular matrix formation in human three-dimensional leiomyoma cultures

Fertil Steril. 2014 Jul;102(1):272-281.e2. doi: 10.1016/j.fertnstert.2014.03.042. Epub 2014 May 10.

Abstract

Objective: To investigate the impact of liarozole on transforming growth factor-β3 (TGF-β3) expression, TGF-β3 controlled profibrotic cytokines, and extracellular matrix formation in a three-dimensional (3D) leiomyoma model system.

Design: Molecular and immunohistochemical analysis in a cell line evaluated in a three-dimensional culture.

Setting: Laboratory study.

Patient(s): None.

Intervention(s): Treatment of leiomyoma and myometrial cells with liarozole and TGF-β3 in a three-dimensional culture system.

Main outcome measure(s): Quantitative real-time reverse-transcriptase polymerase chain reaction and Western blotting to assess fold gene and protein expression of TGF-β3 and TGF-β3 regulated fibrotic cytokines: collagen 1A1 (COL1A1), fibronectin, and versican before and after treatment with liarozole, and confirmatory immunohistochemical stains of treated three-dimensional cultures.

Result(s): Both TGF-β3 gene and protein expression were elevated in leiomyoma cells compared with myometrium in two-dimensional and 3D cultures. Treatment with liarozole decreased TGF-β3 gene and protein expression. Extracellular matrix components versican, COL1A1, and fibronectin were also decreased by liarozole treatment in 3D cultures. Treatment of 3D cultures with TGF-β3 increased gene expression and protein production of COL1A1, fibronectin, and versican.

Conclusion(s): Liarozole decreased TGF-β3 and TGF-β3-mediated extracellular matrix expression in a 3D uterine leiomyoma culture system.

Keywords: Extracellular matrix; leiomyoma; liarozole; three-dimensional model; transforming growth factor-beta.

MeSH terms

  • Antineoplastic Agents, Hormonal / pharmacology*
  • Cell Culture Techniques
  • Collagen Type I / genetics
  • Collagen Type I / metabolism
  • Collagen Type I, alpha 1 Chain
  • Down-Regulation
  • Extracellular Matrix / metabolism*
  • Female
  • Fibronectins / genetics
  • Fibronectins / metabolism
  • Humans
  • Imidazoles / pharmacology*
  • Leiomyoma / genetics
  • Leiomyoma / metabolism*
  • Leiomyoma / pathology
  • Myometrium / drug effects*
  • Myometrium / metabolism
  • Myometrium / pathology
  • Transforming Growth Factor beta3 / genetics
  • Transforming Growth Factor beta3 / metabolism*
  • Tumor Cells, Cultured
  • Uterine Neoplasms / genetics
  • Uterine Neoplasms / metabolism*
  • Uterine Neoplasms / pathology
  • Versicans / genetics
  • Versicans / metabolism

Substances

  • Antineoplastic Agents, Hormonal
  • Collagen Type I
  • Collagen Type I, alpha 1 Chain
  • Fibronectins
  • Imidazoles
  • TGFB3 protein, human
  • Transforming Growth Factor beta3
  • VCAN protein, human
  • Versicans
  • liarozole