Leiomyoma cells in 3-dimensional cultures demonstrate an attenuated response to fasudil, a Rho-kinase inhibitor, when compared to 2-dimensional cultures

Minnie Malik, Joy Britten, James Segars, William H. Catherino

Research output: Contribution to journalArticle

Abstract

Uterine leiomyomata are common benign tumors in women of reproductive age and demonstrate an attenuated response to mechanical signaling that involves Rho and integrins. To further characterize the impairment in Rho signaling, we studied the effect of Rho-kinase inhibitor, fasudil, on extracellular matrix production, in 2-dimensional (2D) and 3-dimensional (3D) cultures of leiomyoma and myometrial cells. Leiomyoma 2D cultures demonstrated a rapid decrease in gene transcripts and protein for fibronectin, procollagen 1A, and versican. In 3D cultures, fibronectin and procollagen 1A proteins demonstrated increased levels at lower concentrations of fasudil, followed by a concentration-dependent decrease. Versican protein increased up to 3-fold, whereas fibromodulin demonstrated a significant decrease of 1.92-fold. Myometrial 2D or 3D cultures demonstrated a decrease in all proteins after 72 hours of treatment. The 3D leiomyoma cultures demonstrated a significant increase in active RhoA, followed by a concentration-dependent decrease at higher concentrations. A concentration-dependent increase in phospho-extracellular regulated signal kinase and proapoptotic protein Bax was observed in 3D leiomyoma cultures. Fasudil relaxed the contraction of the 3D collagen gels caused by myometrium and leiomyoma cell growth. These findings indicate that the altered state of Rho signaling in leiomyoma was more clearly observed in 3D cultures. The results also suggest that fasudil may have clinical applicability for treatment of uterine leiomyoma.

Original languageEnglish (US)
Pages (from-to)1126-1138
Number of pages13
JournalReproductive Sciences
Volume21
Issue number9
DOIs
StatePublished - Sep 1 2014

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Keywords

  • 2D and 3D cultures
  • RhoA
  • extracellular matrix
  • fasudil
  • gel contraction
  • uterine leiomyoma

ASJC Scopus subject areas

  • Obstetrics and Gynecology

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