Protein kinase C delta and eta isoenzymes control the shedding of the interleukin 6 receptor alpha in myeloma cells

Biochem J. 2001 Aug 15;358(Pt 1):193-200. doi: 10.1042/0264-6021:3580193.

Abstract

The soluble interleukin 6 receptor alpha is an agonistic molecule of interleukin 6 (IL-6) and is important in the biology of multiple myeloma. More precisely, it potentiates the deleterious effects of IL-6 during tumour progression, facilitating angiogenesis and bone resorption. Because the mechanisms involved in the shedding of the interleukin 6 receptor alpha (IL-6Ralpha) in multiple myeloma are not known, we have investigated them in the XG-6 human myeloma cell line. Here we provide evidence that PMA-induced IL-6Ralpha shedding is controlled by a metalloproteinase and by protein kinase C (PKC) isoenzymes that do not require Ca(2+) for their activation. We show that XG-6 cells express PKC-delta, -eta and -zeta isoenzymes. However, after stimulation with PMA, only PKC-delta and PKC-eta are activated, as shown by their translocation to the membrane. Treatment with PMA induces an increase in PKC-delta phosphorylation in its active loop. In addition, by using rottlerin, a specific inhibitor of PKC-delta, we demonstrate that PKC-delta is involved in the PMA-induced shedding of IL-6Ralpha. With the use of UO126, a specific inhibitor of the mitogen-activated protein kinase (MAPK) pathway, we show that the PMA-induced IL-6Ralpha shedding is mediated in part by the MAPK pathway. Finally, whereas GF109203X, a general PKC inhibitor, inhibits the activation of ERK1/2 (extracellular signal-regulated protein kinase 1/2), rottlerin has no inhibitory effect, indicating that the Ras/MAPK activation is PKC-dependent but PKC-delta-independent. Taken together, these results suggest that the PMA-induced shedding of IL-6Ralpha is mediated by a PKC isoenzyme network.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetophenones / pharmacology
  • Antibodies, Monoclonal / metabolism
  • Benzopyrans / pharmacology
  • Blotting, Western
  • Calcium / metabolism
  • Down-Regulation
  • Enzyme Activation
  • Enzyme Inhibitors / pharmacology
  • Flow Cytometry
  • Humans
  • Isoenzymes / physiology*
  • MAP Kinase Signaling System
  • Microscopy, Fluorescence
  • Multiple Myeloma / metabolism*
  • Neovascularization, Pathologic
  • Phosphorylation
  • Protein Isoforms
  • Protein Kinase C / physiology*
  • Protein Kinase C-delta
  • Protein Transport
  • Receptors, Interleukin-6 / metabolism*
  • Subcellular Fractions
  • Tetradecanoylphorbol Acetate
  • Tumor Cells, Cultured

Substances

  • Acetophenones
  • Antibodies, Monoclonal
  • Benzopyrans
  • Enzyme Inhibitors
  • Isoenzymes
  • Protein Isoforms
  • Receptors, Interleukin-6
  • rottlerin
  • protein kinase C eta
  • PRKCD protein, human
  • Protein Kinase C
  • Protein Kinase C-delta
  • Tetradecanoylphorbol Acetate
  • Calcium