Blockade of mitogen-activated protein kinase cascade signaling in interleukin 6-independent multiple myeloma cells

Clin Cancer Res. 1997 Jun;3(6):1017-22.

Abstract

Interleukin 6 (IL-6) is a growth factor for multiple myeloma (MM) cells, yet not all MM cell lines or patient cells require IL-6 for their growth. It is well known that IL-6 activates the signal transducers and activators of transcription (stat) 1-stat3 heterodimer, stat3 homodimer, and Ras-dependent mitogen-activated protein kinase (MAPK) cascades in multiple cell systems. We have shown previously that the MAPK pathway is an important pathway for IL-6-mediated MM cell growth. In this study, we delineate the pattern of upstream MAPK cascade activation in IL-6-responsive B9 cells and in IL-6-nonresponsive U266, OCI-My5, and RPMI8226 MM cells to define sites of blockade of this pathway associated with loss of responsiveness to IL-6. In B9 cells, IL-6 triggered the following in sequence: gp130 phosphorylation, gp130-to-protein tyrosine phosphatase 1D (PTP1D) binding, PTP1D phosphorylation, PTP1D complex formation with Grb2-Son of sevenless 1 (Sos1), and Sos1 phosphorylation. gp130 phosphorylation, gp130-to-PTP1D binding, PTP1D phosphorylation, and PTP1D-to-Grb2 binding are also induced by IL-6 in all IL-6-independent MM cell lines studied. However, Grb2 is not associated with Sos1, and neither Grb2-to-Sos1 binding nor Sos1 phosphorylation is triggered by IL-6 in OCI-My5 MM cells. On the other hand, Grb2 and Sos1 are associated constitutively in U266 and RPMI8226 MM cells, but phosphorylation of Sos1 is not induced by IL-6. These data suggest that lack of Sos1 activation is associated with loss of IL-6 responsiveness in MM cell lines that grow independently of IL-6.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Animals
  • Calcium-Calmodulin-Dependent Protein Kinases / antagonists & inhibitors
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
  • Cell Division
  • GRB2 Adaptor Protein
  • Guanine Nucleotide Exchange Factors
  • Humans
  • Interleukin-6 / pharmacology*
  • Interleukin-6 / physiology
  • Mice
  • Multiple Myeloma
  • Phosphorylation
  • Protein Tyrosine Phosphatases / metabolism
  • Proteins / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction / physiology*
  • Transfection
  • Tumor Cells, Cultured
  • ras Guanine Nucleotide Exchange Factors

Substances

  • Adaptor Proteins, Signal Transducing
  • GRB2 Adaptor Protein
  • GRB2 protein, human
  • Grb2 protein, mouse
  • Guanine Nucleotide Exchange Factors
  • Interleukin-6
  • Proteins
  • Recombinant Fusion Proteins
  • ras Guanine Nucleotide Exchange Factors
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Protein Tyrosine Phosphatases