Interleukin-1beta stimulates placental leucine aminopeptidase/oxytocinase expression in BeWo choriocarcinoma cells

Mol Hum Reprod. 2003 Feb;9(2):103-10. doi: 10.1093/molehr/gag015.

Abstract

In addition to prostaglandins, inflammatory cytokines induce uterine contraction via oxytocin (OT). Placental leucine aminopeptidase (P-LAP), an oxytocinase that is identical to cystine aminopeptidase, destroys OT activity. Patients with spontaneous preterm delivery have higher concentrations of inflammatory cytokines and lower P-LAP activities than those with normal delivery. In addition, the P-LAP promoter region contains putative binding sites for cytokine-induced transcription factors. We therefore postulated that inflammatory cytokines suppress P-LAP expression and examined this notion using BeWo choriocarcinoma cells cultured in the presence of cytokines. However, interleukin-1beta (IL-1beta) increased P-LAP activity in a time- and dose-dependent manner. Furthermore, Western blot analysis showed a dose-dependent increase of P-LAP proteins. We also detected IL-1 type I receptor mRNA in BeWo cells by RT-PCR. Semi-quantitative RT-PCR and Southern blot analysis showed that IL-1beta also increased P-LAP mRNA, which was abrogated by prior exposure to cycloheximide. Luciferase assays did not reveal any regulatory regions that could explain IL-1beta-induced P-LAP mRNA accumulation within 1.1 kb upstream of the P-LAP gene. Immunohistochemical analysis of human placenta with chorioamnionitis demonstrated prominent P-LAP staining at sites of abundant inflammatory cell infiltration. These findings indicated that prolonged exposure to IL-1beta induces P-LAP in the trophoblasts, possibly via other de-novo protein synthesis, which contradicted our initial hypothesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Choriocarcinoma
  • Cycloheximide / pharmacology
  • Cystinyl Aminopeptidase / drug effects
  • Cystinyl Aminopeptidase / genetics*
  • Cystinyl Aminopeptidase / metabolism
  • Female
  • Gene Expression Regulation, Enzymologic / drug effects
  • Gene Expression Regulation, Neoplastic / drug effects
  • Genes, Reporter
  • Humans
  • Interleukin-1 / pharmacology*
  • Luciferases / genetics
  • Pregnancy
  • RNA, Messenger / genetics
  • Receptors, Interleukin-1 / genetics
  • Recombinant Fusion Proteins / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription, Genetic / drug effects
  • Transfection
  • Tumor Cells, Cultured
  • Uterine Neoplasms

Substances

  • Interleukin-1
  • RNA, Messenger
  • Receptors, Interleukin-1
  • Recombinant Fusion Proteins
  • Cycloheximide
  • Luciferases
  • Cystinyl Aminopeptidase
  • leucyl-cystinyl aminopeptidase