Cytosolic phospholipase A2 Group IValpha but not secreted phospholipase A2 Group IIA, V, or X induces interleukin-8 and cyclooxygenase-2 gene and protein expression through peroxisome proliferator-activated receptors gamma 1 and 2 in human lung cells

J Biol Chem. 2004 Nov 19;279(47):48550-61. doi: 10.1074/jbc.M408926200. Epub 2004 Aug 25.

Abstract

It has been reported that interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2) expression is regulated by peroxisome proliferator-activated receptor (PPAR)-gamma synthetic ligands. We have shown previously that cytosolic phospholipase A2 (cPLA2) is able to activate gene expression through PPAR-gamma response elements (Pawliczak, R., Han, C., Huang, X. L., Demetris, A. J., Shelhamer, J. H., and Wu, T. (2002) J. Biol. Chem. 277, 33153-33163). In this study we investigated the influence of cPLA2 and secreted phospholipase A2 (sPLA2) Group IIA, Group V, and Group X on IL-8 and COX-2 expression in human lung epithelial cells (A549 cells). We also studied the results of cPLA2 activation by epidermal growth factor (EGF) and calcium ionophore (A23187) on IL-8 and COX-2 reporter gene activity, mRNA level, and protein synthesis. cPLA2 overexpression and activation increased both IL-8 and COX-2 reporter gene activity. Overexpression and activation of Group IIA, Group V, or Group X sPLA2s did not increase IL-8 and COX-2 reporter gene activity. Methyl arachidonyl fluorophosphate, a cPLA2 inhibitor, inhibited the effect of A23187 and of EGF on both IL-8 and COX-2 reporter gene activity, steady state levels of IL-8 and COX-2 mRNA, and IL-8 and COX-2 protein expression. Small inhibitory RNAs directed against PPAR-gamma1 and -gamma2 blunted the effect of A23187 and of EGF on IL-8 and COX-2 protein expression. Moreover small inhibitory RNAs directed against cPLA2 decreased the effect of A23187 and EGF on IL-8 and COX-2 protein expression. These results demonstrate that cPLA2 has an influence on IL-8 and COX 2 gene and protein expression at least in part through PPAR-gamma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arachidonic Acid / metabolism
  • Calcimycin / pharmacology
  • Cell Line, Tumor
  • Cell Nucleus / metabolism
  • Cyclooxygenase 2
  • Cytosol / enzymology
  • Epidermal Growth Factor / pharmacology
  • Gene Expression Regulation
  • Genes, Reporter
  • Group II Phospholipases A2
  • Group IV Phospholipases A2
  • Group V Phospholipases A2
  • Group X Phospholipases A2
  • Humans
  • Immunoblotting
  • Interleukin-8 / metabolism*
  • Ionophores / pharmacology
  • Isoenzymes / metabolism*
  • Lung / metabolism*
  • Membrane Proteins
  • PPAR gamma / metabolism*
  • Phospholipases / metabolism
  • Phospholipases A / physiology*
  • Phospholipases A2
  • Plasmids / metabolism
  • Prostaglandin-Endoperoxide Synthases / metabolism*
  • RNA / metabolism
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors
  • Transfection

Substances

  • Interleukin-8
  • Ionophores
  • Isoenzymes
  • Membrane Proteins
  • PPAR gamma
  • RNA, Messenger
  • RNA, Small Interfering
  • Arachidonic Acid
  • Calcimycin
  • Epidermal Growth Factor
  • RNA
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • Prostaglandin-Endoperoxide Synthases
  • Phospholipases
  • Phospholipases A
  • Group II Phospholipases A2
  • Group IV Phospholipases A2
  • Group V Phospholipases A2
  • Group X Phospholipases A2
  • PLA2G10 protein, human
  • PLA2G4A protein, human
  • PLA2G5 protein, human
  • Phospholipases A2