IL-1 beta- and IL-4-induced down-regulation of autotaxin mRNA and PC-1 in fibroblast-like synoviocytes of patients with rheumatoid arthritis (RA).
Autotaxin (ATX) is a 125-kD ectonucleotide pyrophosphate/phosphodiesterase, which was initially isolated and cloned from human melanoma cells as a potent stimulator of tumour cell motility. ATX shows 44% identity to the plasma cell membrane marker PC-1. Recently, we described the decreased expression of ATX mRNA in cultured fibroblast-like synoviocytes (SFC) of patients with RA by interferon-gamma. In this study using a competitive reverse transcriptase-polymerase chain reaction, we show an increased ATX mRNA expression in SFC from patients with RA in comparison with synoviocytes from non-RA patients. The median ATX mRNA amount in SFC of RA patients (440 pg/microg total RNA) was five-fold higher than the expression in synoviocytes from non-RA patients (80 pg/microg total RNA) or foreskin fibroblasts (MRHF cells, 90 pg/microg total RNA). In contrast to the elevated ATX mRNA expression in SFC of patients with RA, we did not measure increased mRNA amounts of PC-1 in these cells. Both the ATX mRNA amount and the 5'-nucleotide phosphodiesterase (PDE) activity of SFC lysate were reduced after treatment of SFC with the cytokines IL-1beta or IL-4. IL-1beta and IL-4 induced a down-regulation of PC-1 mRNA and protein expression in SFC. In SFC treated with transforming growth factor-beta the expression of PC-1 mRNA and protein was increased, whereas no significant effect on ATX mRNA expression was detectable. Pharmacological drugs used in therapy for RA, such as dexamethasone, cyclosporin, methotrexate and indomethacin, did not show a statistically significant effect on either ATX mRNA or PC-1 mRNA expression. Only pentoxifylline suppressed ATX mRNA as well as PC-1 mRNA expression. In conclusion, we show a tight regulation of ATX and PC-1 gene expression by cytokines detectable in the inflamed tissue of RA. Further investigations will deal with the regulation of ATX protein expression as well as with the function of ATX in RA.
Kehlen
A
A
Institute of Medical Immunology and Institute of Physiological Chemistry, Martin Luther University, Halle, Germany. astrid.kehlen@medizin.uni-halle.de
Lauterbach
R
R
Santos
A N
AN
Thiele
K
K
Kabisch
U
U
Weber
E
E
Riemann
D
D
Langner
J
J
D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D001172:Arthritis, Rheumatoid / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D002478:Cells, Cultured; D016207:Cytokines / Q000502:physiology; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D004789:Enzyme Activation / Q000276:immunology; D005347:Fibroblasts / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology; D005956:Glucose-6-Phosphate Isomerase / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D006023:Glycoproteins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007375:Interleukin-1 / Q000502:physiology*; D015847:Interleukin-4 / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D009097:Multienzyme Complexes; D043264:Phosphodiesterase I; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D011755:Pyrophosphatases; D012333:RNA, Messenger / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D013256:Steroids; D013583:Synovial Membrane / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D000099189:Lysophospholipase D
D000893:Anti-Inflammatory Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D016207:Cytokines; D005956:Glucose-6-Phosphate Isomerase; D006023:Glycoproteins; D007375:Interleukin-1; D015847:Interleukin-4; D008562:Membrane Glycoproteins; D009097:Multienzyme Complexes; D043264:Phosphodiesterase I; D010726:Phosphodiesterase Inhibitors; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases; D012333:RNA, Messenger; D013256:Steroids; D000099189:Lysophospholipase D; C065962:ectonucleotide pyrophosphatase phosphodiesterase 1