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


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


METHODS
Laboratory study.


METHODS
None.


METHODS
Treatment of leiomyoma and myometrial cells with liarozole and TGF-β3 in a three-dimensional culture system.


METHODS
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.


RESULTS
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.


CONCLUSIONS
Liarozole decreased TGF-β3 and TGF-β3-mediated extracellular matrix expression in a 3D uterine leiomyoma culture system.
Program in Reproductive and Adult Endocrinology Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland; Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland.
Gary
G
Levy
Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland.
Minnie
M
Malik
Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland.
Joy
J
Britten
Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland.
Melissa
M
Gilden
Program in Reproductive and Adult Endocrinology Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland; Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland.
James
J
Segars
Program in Reproductive and Adult Endocrinology Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland; Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland. Electronic address: william.catherino@usuhs.edu.
William H
WH
Catherino
D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology*; D018929:Cell Culture Techniques; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D000090602:Collagen Type I, alpha 1 Chain; D015536:Down-Regulation; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D007889:Leiomyoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D009215:Myometrium / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D053782:Transforming Growth Factor beta3 / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D053675:Versicans / Q000235:genetics / Q000378:metabolism
D018931:Antineoplastic Agents, Hormonal; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D005353:Fibronectins; D007093:Imidazoles; C509044:TGFB3 protein, human; D053782:Transforming Growth Factor beta3; C508322:VCAN protein, human; D053675:Versicans; C061121:liarozole
Extracellular matrix; leiomyoma; liarozole; three-dimensional model; transforming growth factor-beta