Effect of estrogen-active compounds on the expression of RACK1 and immunological implications

Arch Toxicol. 2020 Jun;94(6):2081-2095. doi: 10.1007/s00204-020-02756-9. Epub 2020 Apr 23.

Abstract

We previously demonstrated the existence of a balance among steroid hormones, i.e. glucocorticoids and androgens, in RACK1 (receptor for activated C kinase 1) expression and innate immunity activation, which may offer the opportunity to use RACK1 expression as marker to evaluate immunotoxicity of hormone-active substances. Because of the existence of close interconnections between the different steroid hormone receptors with overlapping ligand specificities and signaling pathways, in this study, we wanted to investigate a possible effect of estrogenic active compounds, namely 17β-estradiol, diethylstilbestrol, and zearalenone, on RACK-1 expression and innate immune responses using THP-1 cells as experimental model. All compounds increased RACK1 transcriptional activity as evaluated by reporter luciferase activity, mRNA expression as assessed by real time-PCR and protein expression by western blot analysis, which paralleled an increase in LPS-induced IL-8, TNF-α production, and CD86 expression, which we previously demonstrated to be dependent on RACK1/PKCβ activation. As the induction of RACK1 expression can be blocked by the antagonist G15, induced by the agonist G1 and by the non-cell permeable 17β-estradiol conjugated with BSA, a role of GPER (previously named GPR30) activation in estrogen-induced RACK1 expression could be demonstrated. In addition, a role of androgen receptor (AR) in RACK1 transcription was also demonstrated by the ability of flutamide, a nonsteroidal antiandrogen, to completely prevent diethylstilbestrol-induced RACK1 transcriptional activity and protein expression. Altogether, our data suggest that RACK1 may represent an interesting target of steroid-active compounds, and its evaluation may offer the opportunity to screen the immunotoxic potential of hormone-active substances.

Keywords: Cytokines; Endocrine-disrupting chemicals; Estrogens; Hormones; Immune system; Signal transduction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cytokines / metabolism
  • Diethylstilbestrol / toxicity*
  • Endocrine Disruptors
  • Estradiol / toxicity*
  • Estrogens / toxicity*
  • Humans
  • Immunity, Innate / drug effects*
  • Macrophages / drug effects*
  • Macrophages / immunology
  • Macrophages / metabolism
  • Monocytes / drug effects*
  • Monocytes / immunology
  • Monocytes / metabolism
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism*
  • Proof of Concept Study
  • Receptors for Activated C Kinase / genetics
  • Receptors for Activated C Kinase / metabolism*
  • Receptors, Androgen / drug effects
  • Receptors, Androgen / metabolism
  • Receptors, Estrogen / metabolism
  • Receptors, G-Protein-Coupled / agonists
  • Receptors, G-Protein-Coupled / metabolism
  • Signal Transduction
  • THP-1 Cells
  • Up-Regulation
  • Zearalenone / toxicity*

Substances

  • AR protein, human
  • Cytokines
  • Endocrine Disruptors
  • Estrogens
  • GPER1 protein, human
  • Neoplasm Proteins
  • RACK1 protein, human
  • Receptors for Activated C Kinase
  • Receptors, Androgen
  • Receptors, Estrogen
  • Receptors, G-Protein-Coupled
  • Estradiol
  • Zearalenone
  • Diethylstilbestrol