Expression of arachidonate platelet-type 12-lipoxygenase in human rheumatoid arthritis type B synoviocytes

FEBS Lett. 1997 Sep 1;414(1):159-64. doi: 10.1016/s0014-5793(97)00904-6.

Abstract

In the present study, we have demonstrated platelet-type 12-lipoxygenase (12-LOX) expression in human rheumatoid arthritis (RA) type B synoviocytes by reverse-transcription polymerase chain reaction (RT-PCR). The presence of 12-LOX mRNA in these cells was revealed by classical RT-PCR analysis using platelet-type 12-LOX cDNA primers and the PCR fragment (246 bp) was purified, amplified and sequenced. By sequence analysis, this fragment was determined to be 100% identical to that from platelet-type 12-LOX cDNA. Immunofluorescence data demonstrate that interleukin-1beta (IL-1beta) increases cellular 12-LOX protein. Other results associate specific inflammatory cytokines with the activity of 12-LOX in human RA type B synoviocytes. IL-1beta increased 12S-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) production (4-fold) and we also observed an increase in 12-HETE production (2.5-fold) after incubation of human RA type B synoviocytes with TNF alpha. In contrast to the action of IL-1beta on 12-HETE synthesis, IL-4 and IL-6 did not enhance 12-HETE production. This is the first demonstration of platelet-type 12-LOX cDNA derived from the mRNA of cultured human RA type B synoviocytes.

MeSH terms

  • 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / biosynthesis*
  • Arachidonate 12-Lipoxygenase / biosynthesis
  • Arachidonate 12-Lipoxygenase / genetics*
  • Arachidonic Acid / metabolism*
  • Arthritis, Rheumatoid / metabolism*
  • Arthritis, Rheumatoid / pathology
  • Blood Platelets / enzymology
  • Calcimycin / pharmacology
  • Cells, Cultured
  • Cloning, Molecular
  • DNA Primers
  • Fluorescent Antibody Technique
  • Gene Expression*
  • Humans
  • Interleukin-1 / pharmacology
  • Interleukin-4 / pharmacology
  • Interleukin-6 / pharmacology
  • Microscopy, Fluorescence
  • Polymerase Chain Reaction
  • RNA, Messenger / analysis
  • Sequence Analysis, DNA
  • Synovial Membrane / enzymology*
  • Synovial Membrane / metabolism
  • Synovial Membrane / pathology
  • Tumor Necrosis Factor-alpha / pharmacology

Substances

  • DNA Primers
  • Interleukin-1
  • Interleukin-6
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha
  • Interleukin-4
  • Arachidonic Acid
  • Calcimycin
  • 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid
  • Arachidonate 12-Lipoxygenase