5-aza-2 '-deoxycitidine inhibits cell proliferation, extracellular matrix formation and Wnt/beta-catenin pathway in human uterine leiomyomas

Autores de IIS La Fe

• Maria Cristina Carbajo García

  Autor

• Ana Corachán García

  Autor

• Marina Segura

  Autor

• Javier Monleón Sancho

  Autor

• Julia Escrig Estrems

  Autor

• Amparo Faus Esteve

  Autor

• Antonio Pellicer Martínez

  Autor

• Irene Cervelló Alcaraz

  Autor

• Hortensia Ferrero Chafer

  Autor

Grupos

• Biología Reproductiva y Bioingeniería en Reproducción Humana

• Medicina Fetal y Reproductiva

Abstract

Background: Uterine leiomyoma is a benign tumor with unclear pathogenesis and inaccurate treatment. This tumor exhibits altered DNA methylation related to disease progression. DNMT inhibitors as 5-aza-2'-deoxycytidine (5-aza-CdR), have been suggested to treat tumors in which DNA methylation is altered. We aimed to evaluate whether DNA methylation reversion with 5-aza-CdR reduces cell proliferation and extracellular matrix (ECM) formation in uterine leiomyoma cells to provide a potential treatment option. Methods: Prospective study using uterine leiomyoma and adjacent myometrium tissues and human uterine leiomyoma primary (HULP) cells (n = 16). In tissues, gene expression was analyzed by qRT-PCR and DNMT activity by ELISA. Effects of 5-aza-CdR treatment on HULP cells were assessed by CellTiter, western blot, and qRT-PCR. Results: DNMT1 gene expression was higher in uterine leiomyoma vs myometrium. Similarly, DNMT activity was greater in uterine leiomyoma and HULP cells (6.5 vs 3.8 OD/h/mg; 211.3 vs 63.7 OD/h/mg, respectively). After 5-azaCdR treatment on HULP cells, cell viability was reduced, significantly so at 10 mu M (85.3%). Treatment with 10 mu M 5-aza-CdR on HULP cells significantly decreased expression of proliferation marker PCNA (FC = 0.695) and of ECM proteins (COLLAGEN I FC = 0.654; PAI-1, FC = 0.654; FIBRONECTIN FC = 0.733). 5-aza-CdR treatment also decreased expression of Wnt/beta-catenin pathway final targets, including WISP1 protein expression (10 pM, FC = 0.699), c-MYC gene expression (2 mu M, FC = 0.745 and 10 mu M, FC = 0.728), and MMP7 gene expression (5 mu M, FC = 0.520 and 10 mu M, FC = 0.577). Conclusions: 5-aza-CdR treatment inhibits cell proliferation, ECM formation, and Wnt/beta-catenin signaling pathway targets in HULP cells, suggesting that DNA methylation inhibition is a viable therapeutic target in uterine leiomyoma.

Keywords

• Uterine leiomyoma; Epigenetics; 5-aza-2 '-deoxycitidine; Cell proliferation; Wnt/beta-catenin pathway