Development of an in vitro test battery for the screening of the receptor-mediated mechanism and the spindle-poison mode of action of estrogenic compounds

Environ Toxicol Pharmacol. 2016 Dec;48:245-252. doi: 10.1016/j.etap.2016.11.005. Epub 2016 Nov 9.

Abstract

The implementation of the REACH regulation has imposed the urgent need of developing alternative testing methods to screen large number of compounds more quickly and at lower costs. In this study, a battery of tests, suitable for reproductive toxicology testing, was developed with the objective of detecting the mechanism of action of estrogenic and xenoestrogenic compounds. With this aim, two compounds known for their estrogenic activity, diethylstilbestrol and 17β-estradiol, were used to set up four different in vitro tests: 1) bovine oocyte in vitro maturation assay, 2) bovine preimplantation embryo in vitro culture assay and 3) MCF-7 and 4) BALB/3T3 cell lines proliferation and cytotoxicity assay, respectively. The results show that this battery of tests allows to identify and to distinguish between two major mechanisms of action of (xeno)estrogenic compounds: the receptor-mediated mechanism and the spindle-poison effect on microtubules polimerization.

Keywords: Blastocyst; Diethylstilbestrol; Estradiol; Estrogen receptor alpha; In vitro oocyte maturation techniques (MeSH); MCF-7cells.

MeSH terms

  • Animal Testing Alternatives / methods*
  • Animals
  • BALB 3T3 Cells
  • Cattle
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Diethylstilbestrol / toxicity
  • Dose-Response Relationship, Drug
  • Embryonic Development / drug effects
  • Estradiol / toxicity
  • Estrogens / toxicity*
  • Estrogens, Non-Steroidal / toxicity
  • Humans
  • In Vitro Oocyte Maturation Techniques
  • MCF-7 Cells
  • Meiosis / drug effects
  • Mice
  • Oocytes / drug effects
  • Oocytes / metabolism
  • Receptors, Estrogen / metabolism*
  • Reproduction / drug effects
  • Spindle Apparatus / drug effects*
  • Toxicity Tests / methods*

Substances

  • Estrogens
  • Estrogens, Non-Steroidal
  • Receptors, Estrogen
  • Estradiol
  • Diethylstilbestrol