EGF-like growth factors induce COX-2-derived PGE2 production through ERK1/2 in human granulosa cells

J Clin Endocrinol Metab. 2013 Dec;98(12):4932-41. doi: 10.1210/jc.2013-2662. Epub 2013 Oct 3.

Abstract

Context: Aberrant regulation of ovulation is one of the major causes of infertility. In animal models, 3 epidermal growth factor (EGF)-like growth factors, amphiregulin (AREG), betacellulin (BTC), and epiregulin (EREG), have been shown to be involved in ovulation by regulating cyclooxygenase-2 (COX-2) expression and prostaglandin E2 (PGE2) production. However, whether the same is true in humans remains largely unknown.

Objective: Our objective was to investigate the effects of AREG, BTC, and EREG on COX-2 expression and PGE2 production in human granulosa cells.

Design and setting: SVOG cells are human granulosa cells that were obtained from women undergoing in vitro fertilization and immortalized with SV40 large T antigen. SVOG cells were used to investigate the effect of AREG, BTC, and EREG on ovulation-related functions at an academic research center.

Main outcome measures: Levels of mRNA and protein were examined by quantitative RT-PCR and Western blotting, respectively. The protein levels of PGE2 were measured by ELISA.

Results: LH treatment upregulated AREG, BTC, EREG, and COX-2. Knockdown of EGF receptor (EGFR) attenuated LH-induced COX-2 expression and PGE2 production. Treatment with AREG, BTC, and EREG upregulated COX-2 expression and PGE2 production. The stimulatory effects of AREG, BTC, and EREG on COX-2 expression and PGE2 production were blocked by inhibition of EGFR activity and expression. AREG-, BTC-, and EREG-activated ERK1/2 signaling, but not Akt signaling, was required for AREG-, BTC-, and EREG-induced COX-2 expression and PGE2 production.

Conclusion: AREG, BTC, and EREG induced PGE2 production by upregulating COX-2 expression through ERK1/2 signaling in human granulosa cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amphiregulin
  • Betacellulin
  • Cell Line, Transformed
  • Cyclooxygenase 2 / chemistry
  • Cyclooxygenase 2 / genetics
  • Cyclooxygenase 2 / metabolism*
  • Dinoprostone / metabolism
  • EGF Family of Proteins
  • Enzyme Induction*
  • Epidermal Growth Factor / genetics
  • Epidermal Growth Factor / metabolism*
  • Epiregulin
  • ErbB Receptors / antagonists & inhibitors
  • ErbB Receptors / genetics
  • ErbB Receptors / metabolism
  • Female
  • Glycoproteins / genetics
  • Glycoproteins / metabolism*
  • Granulosa Cells / enzymology
  • Granulosa Cells / metabolism*
  • Humans
  • Intercellular Signaling Peptides and Proteins / genetics
  • Intercellular Signaling Peptides and Proteins / metabolism*
  • Luteinizing Hormone / metabolism
  • MAP Kinase Signaling System*
  • RNA Interference
  • RNA, Messenger
  • RNA, Small Interfering
  • Recombinant Proteins / metabolism
  • Up-Regulation

Substances

  • AREG protein, human
  • Amphiregulin
  • BTC protein, human
  • Betacellulin
  • EGF Family of Proteins
  • EREG protein, human
  • Epiregulin
  • Glycoproteins
  • Intercellular Signaling Peptides and Proteins
  • RNA, Messenger
  • RNA, Small Interfering
  • Recombinant Proteins
  • Epidermal Growth Factor
  • Luteinizing Hormone
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • EGFR protein, human
  • ErbB Receptors
  • Dinoprostone