Mechanism of toxic action of 2,3,7,8-tetrachlorodibenzo-P-dioxin (TCDD) in cultured human luteinized granulosa cells

Essam Enan, Francisco Moran, Catherine A. VandeVoort, Dennis R. Stewart, James W. Overstreet, Bill L. Lasley

Research output: Contribution to journalArticle

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Abstract

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) caused a significant decrease in estradiol (E2) production when it was administered to human luteinized granulosa cells (hLGCs) in culture. We investigated the involvement of the epidermal growth factor receptor (EGFR) and protein tyrosine kinase (PTK) in this TCDD-induced toxicity. Upregulation in 125I-EGF binding to EGFR was measured after 24 h of TCDD treatment, while downregulation in EGFR binding was measured after 72 h of TCDD treatment. Upregulation of EGFR binding was associated with a significant decrease in postnuclear (7000 x g supernatant) PTK activity, but this activity was stimulated after 72 h of TCDD treatment. TCDD altered the level of tyrosine phosphorylation in proteins with molecular weights 35, 40, 43, 45, 60, and > 205 kDa. TCDD caused a significant increase in postnuclear cAMP-dependent protein kinase (PKA) after 24 h of treatment. The actions of TCDD on protein kinases were partially blocked by the protein synthesis inhibitor, cycloheximide. On the other hand, TCDD increased nuclear PTK and decreased nuclear PKA activity. E2 inhibited the postnuclear and nuclear activity of both PTK and PKA in control samples, but did not affect TCDD actions on either postnuclear or nuclear PTK activity. However, E2 abolished the stimulatory effect of TCDD on PKA activity in postnuclear protein. In the presence of insulin, TCDD did not induce any additional changes in postnuclear or nuclear PTK. Forskolin (FK) alone inhibited postnuclear PTK activity and stimulated its nuclear activity. The addition of TCDD 20 min after FK resulted in an increase in postnuclear PTK, but there was little change in nuclear PTK as compared to the effect of FK alone. The stimulatory effect of TCDD on postnuclear PKA activity was enhanced by insulin and TCDD reversed the negative effect of FK, but there was no effect of either insulin or FK on the inhibition by TCDD of nuclear PKA activity. TCDD decreased the activity of MAP2 kinase and reduced the binding activity of AP-1 DNA when given alone, and also blocked the E2 stimulation of MAP2K. These findings suggest that TCDD may interrupt the endocrine function of hLGCs through the blockage of the mitotic signal directly or indirectly through the interaction of PTK/MAP2K and PKA signaling.

Original languageEnglish (US)
Pages (from-to)497-508
Number of pages12
JournalReproductive Toxicology
Volume10
Issue number6
DOIs
StatePublished - Nov 1996

Fingerprint

Toxic Actions
Dioxins
Granulosa Cells
Protein-Tyrosine Kinases
Protein Kinases
Nuclear Proteins
Colforsin
Epidermal Growth Factor Receptor
Insulin
Focal Adhesion Kinase 1
Up-Regulation
Phosphorylation
Protein Synthesis Inhibitors

Keywords

  • activator protein-1 (AP-1)
  • EGFR
  • human luteinized granulosa cells
  • protein phosphorylation
  • TCDD

ASJC Scopus subject areas

  • Toxicology

Cite this

Mechanism of toxic action of 2,3,7,8-tetrachlorodibenzo-P-dioxin (TCDD) in cultured human luteinized granulosa cells. / Enan, Essam; Moran, Francisco; VandeVoort, Catherine A.; Stewart, Dennis R.; Overstreet, James W.; Lasley, Bill L.

In: Reproductive Toxicology, Vol. 10, No. 6, 11.1996, p. 497-508.

Research output: Contribution to journalArticle

Enan, Essam ; Moran, Francisco ; VandeVoort, Catherine A. ; Stewart, Dennis R. ; Overstreet, James W. ; Lasley, Bill L. / Mechanism of toxic action of 2,3,7,8-tetrachlorodibenzo-P-dioxin (TCDD) in cultured human luteinized granulosa cells. In: Reproductive Toxicology. 1996 ; Vol. 10, No. 6. pp. 497-508.
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