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

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.

Gonadotropin-releasing hormone analogues inhibit leiomyoma extracellular matrix despite presence of gonadal hormones

“…One major advantage of the 3D leiomyoma in vitro cell culture model system has been the capability of staining the matrix for ECM proteins, as we have demonstrated previously (70). The effect of GnRH-a and gonadal hormones on FN1 and VCAN protein production by leiomyoma cells was clearly depicted by an increase or loss of staining of the proteins as analyzed by immunostaining.…”

Fucoidan Inhibits the Proliferation of Leiomyoma Cells and Decreases Extracellular Matrix-Associated Protein Expression

“…Similarly, Levy et al demonstrated that untreated leiomyoma showed a 2-fold increase in fibronectin protein concentration compared with untreated myometrial cells. Additionally, TGFβ3 treatment enhanced the expression of fibronectin in both myometrial and leiomyoma cells [40]. Earlier studies demonstrated that collagen type 1 (COL1A1) protein production was higher in untreated leiomyoma than myometrial cells, while Joseph et al demonstrated that elevating TGFβ3 concentration of 0.1 ng/mL to 10 ng/mL resulted in concentration-dependent increase in COL1A1 in both myometrial and leiomyoma cells [41].…”

Signaling Pathways in Leiomyoma: Understanding Pathobiology and Implications for Therapy

“…Therefore, it can be a potential therapeutic agent. Also, Levy and colleagues demonstrated that liarozole, an agent that modulates RA metabolism, can inhibit ECM formation in leiomyoma 3-D culture by inhibiting TGF-β3 expression (157).…”