Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Manuscript
  • Published:

Analysis of IL-6-mediated growth control of myeloma cells using a gp130 chimeric receptor approach

Abstract

Interleukin 6 (IL-6) has been shown to be a key growth factor for myeloma cells. To study IL-6 signal transduction in multiple myeloma (MM), we employed chimeric receptors composed of the epidermal growth factor receptor (EGFR) extracellular domain, gp130 transmembrane domain, and full-length or truncated gp130 cytoplasmic domains lacking regions previously shown to be necessary for MAPK, STAT1, and STAT3 activation. The IL-6-dependent KAS-6/1 MM cell line was transfected with various chimeric receptor constructs and assayed for EGF responsiveness. EGF stimulation surprisingly stimulated DNA synthesis in all transfectants, regardless of receptor length. When cell proliferation was assayed instead, only transfectants capable of inducing high levels of STAT3 activation proliferated in response to EGF. Additional studies revealed that EGF stimulation resulted in tyrosine phosphorylation of endogenous gp130 in cells expressing the chimeric receptor. Replacing the gp130 transmembrane region with the EGFR transmembrane domain diminished but did not disrupt this interaction. This receptor interaction was also observed in the IL-6-dependent MM cell line ANBL-6. In summary, although our results suggest that STAT activation is crucial in gp130-mediated proliferation of myeloma cells, these results must be interpreted with caution given our demonstration of the interaction between chimeric and endogenous receptors in myeloma cells. Importantly, this interaction has not been noted in studies utilizing the same gp130 chimeric receptor system in non-MM cells.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Hallek M, Leif Bergsagel P, Anderson KC . Multiple myeloma: increasing evidence for a multistep transformation process Blood 1998 91: 3–21

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Zhang XG, Gaillard JP, Robillard N, Lu ZY, Gu ZJ, Jourdan M, Boiron JM, Bataille R, Klein B . Reproducible obtaining of human myeloma cell lines as a model for tumor stem cell study in human multiple myeloma Blood 1994 83: 3654–3663

    CAS  PubMed  Google Scholar 

  3. Westendorf JJ, Ahmann GJ, Greipp PR, Witzig TE, Lust JA, Jelinek DF . Establishment and characterization of three myeloma cell lines that demonstrate variable cytokine responses and abilities to produce autocrine interleukin-6 Leukemia 1996 10: 866–876

    CAS  PubMed  Google Scholar 

  4. Taga T, Kishimoto T . Gp130 and the interleukin-6 family of cytokines Ann Rev Immunol 1997 15: 797–819

    Article  CAS  Google Scholar 

  5. Bennett AM, Tang TL, Sugimoto S, Walsh CT, Neel BG . Protein-tyrosine-phosphatase SHPTP2 couples platelet-derived growth factor receptor beta to Ras Proc Natl Acad Sci USA 1994 91: 7335–7339

    Article  CAS  Google Scholar 

  6. Lee IS, Liu Y, Narazaki M, Hibi M, Kishimoto T, Taga T . Vav is associated with signal transducing molecules gp130, Grb2 and Erk2, and is tyrosine phosphorylated in response to interleukin-6 FEBS Lett 1997 401: 133–137

    Article  CAS  Google Scholar 

  7. Nishida K, Yoshida Y, Itoh M, Fukada T, Ohtani T, Shirogane T, Atsumi T, Takahashi-Tezuka M, Ishihara K, Hibi M . Gab-family adapter proteins act downstream of cytokine and growth factor receptors and T- and B-cell antigen receptors Blood 1999 93: 1809–1816

    CAS  PubMed  Google Scholar 

  8. Takahashi-Tezuka M, Yoshida Y, Fukada T, Ohtani T, Yamanaka Y, Nishida K, Nakajima K, Hibi M, Hirano T . Gab1 acts as an adapter molecule linking the cytokine receptor gp130 to ERK mitogen-activated protein kinase Mol Cell Biol 1998 18: 4109–4117

    Article  CAS  Google Scholar 

  9. Yamanaka Y, Nakajima K, Fukada T, Hibi M, Hirano T . Differentiation and growth arrest signals are generated through the cytoplasmic region of gp130 that is essential for Stat3 activation EMBO J 1996 15: 1557–1565

    Article  CAS  Google Scholar 

  10. Stahl N, Farruggella TJ, Boulton TG, Zhong Z, Darnell JE Jr, Yancopoulos GD . Choice of STATs and other substrates specified by modular tyrosine-based motifs in cytokine receptors Science 1995 267: 1349–1353

    Article  CAS  Google Scholar 

  11. Berger LC, Hawley TS, Lust JA, Goldman SJ, Hawley RG . Tyrosine phosphorylation of JAK-TYK kinases in malignant plasma cell lines growth-stimulated by interleukins 6 and 11 Biochem Biophys Res Commun 1994 202: 596–605

    Article  CAS  Google Scholar 

  12. Ogata A, Chauhan D, Teoh G, Treon SP, Urashima M, Schlossman RL, Anderson KC . IL-6 triggers cell growth via the Ras-dependent mitogen-activated protein kinase cascade J Immunol 1997 159: 2212–2221

    CAS  PubMed  Google Scholar 

  13. Catlett-Falcone R, Landowski TH, Oshiro MM, Turkson J, Levitzki A, Savino R, Ciliberto G, Moscinski L, Fernandez-Luna JL, Nunez G . Constitutive activation of Stat3 signaling confers resistance to apoptosis in human U266 myeloma cells Immunity 1999 10: 105–115

    Article  CAS  Google Scholar 

  14. Kumar G, Gupta S, Wang S, Nel AE . Involvement of Janus kinases, p52shc, Raf-1, and MEK-1 in the IL-6-induced mitogen-activated protein kinase cascade of a growth-responsive B cell line J Immunol 1994 153: 4436–4447

    CAS  PubMed  Google Scholar 

  15. Hallek M, Neumann C, Schaffer M, Danhauser-Riedl S, von Bubnoff N, de Vos G, Druker BJ, Yasukawa K, Griffin JD, Emmerich B . Signal transduction of interleukin-6 involves tyrosine phosphorylation of multiple cytosolic proteins and activation of Src-family kinases Fyn, Hck, and Lyn in multiple myeloma cell lines Exp Hematol 1997 25: 1367–1377

    CAS  PubMed  Google Scholar 

  16. Neumann C, Zehentmaier G, Danhauser-Riedl S, Emmerich B, Hallek M . Interleukin-6 induces tyrosine phosphorylation of the Ras activating protein Shc, and its complex formation with Grb2 in the human multiple myeloma cell line LP-1 Eur J Immunol 1996 26: 379–384

    Article  CAS  Google Scholar 

  17. Berger LC, Hawley RG . Interferon-beta interrupts interleukin-6-dependent signaling events in myeloma cells Blood 1997 89: 261–271

    CAS  PubMed  Google Scholar 

  18. Ogata A, Chauhan D, Urashima M, Teoh G, Treon SP, Anderson KC . Blockade of mitogen-activated protein kinase cascade signaling in interleukin 6-independent multiple myeloma cells Clin Cancer Res 1997 3: 1017–1022

    CAS  PubMed  Google Scholar 

  19. De Vos J, Jourdan M, Tarte K, Jasmin C, Klein B . JAK2 tyrosine kinase inhibitor tyrphostin AG490 downregulates the mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription (STAT) pathways and induces apoptosis in myeloma cells Br J Haematol 2000 109: 823–828

    Article  CAS  Google Scholar 

  20. Lai CF, Ripperger J, Morella KK, Wang Y, Gearing DP, Fey GH, Baumann H . Separate signaling mechanisms are involved in the control of STAT protein activation and gene regulation via the interleukin 6 response element by the box 3 motif of gp130 J Biol Chem 1995 270: 14847–14850

    Article  CAS  Google Scholar 

  21. Schmitz J, Dahmen H, Grimm C, Gendo C, Muller-Newen G, Heinrich PC, Schaper F . The cytoplasmic tyrosine motifs in full-length glycoprotein 130 have different roles in IL-6 signal transduction J Immunol 2000 164: 848–854

    Article  CAS  Google Scholar 

  22. Kim H, Baumann H . Dual signaling role of the protein tyrosine phosphatase SHP-2 in regulating expression of acute-phase plasma proteins by interleukin-6 cytokine receptors in hepatic cells Mol Cell Biol 1999 19: 5326–5338

    Article  CAS  Google Scholar 

  23. Baumann H, Gearing D, Ziegler SF . Signaling by the cytoplasmic domain of hematopoietin receptors involves two distinguishable mechanisms in hepatic cells J Biol Chem 1994 269: 16297–16304

    CAS  PubMed  Google Scholar 

  24. Fukada T, Hibi M, Yamanaka Y, Takahashi-Tezuka M, Fujitani Y, Yamaguchi T, Nakajima K, Hirano T . Two signals are necessary for cell proliferation induced by a cytokine receptor gp130: involvement of STAT3 in anti-apoptosis Immunity 1996 5: 449–460

    Article  CAS  Google Scholar 

  25. Faris M, Kokot N, Stahl N, Nel AE . Involvement of Stat3 in interleukin-6-induced IgM production in a human B-cell line Immunol 1997 90: 350–357

    Article  CAS  Google Scholar 

  26. Murakami M, Narazaki M, Hibi M, Yawata H, Yasukawa K, Hamaguchi M, Taga T, Kishimoto T . Critical cytoplasmic region of the interleukin 6 signal transducer gp130 is conserved in the cytokine receptor family Proc Natl Acad Sci USA 1991 88: 11349–11353

    Article  CAS  Google Scholar 

  27. Jelinek DF, Ahmann GJ, Greipp PR, Jalal SM, Westendorf JJ, Katzmann JA, Kyle RA, Lust JA . Coexistence of aneuploid subclones within a myeloma cell line that exhibits clonal immunoglobulin gene rearrangement: clinical implications Cancer Res 1993 53: 5320–5327

    CAS  PubMed  Google Scholar 

  28. Horton RM, Hunt HD, Ho SN, Pullen JK, Pease LR . Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension Gene 1989 77: 61–68

    Article  CAS  Google Scholar 

  29. Simpson RJ, Hammacher A, Smith DK, Matthews JM, Ward LD . Interleukin-6: structure–function relationships Protein Sci 1997 6: 929–955

    Article  CAS  Google Scholar 

  30. Grotzinger J, Kernebeck T, Kallen KJ, Rose-John S . IL-6 type cytokine receptor complexes: hexamer, tetramer or both? Biol Chem 1999 380: 803–813

    Article  CAS  Google Scholar 

  31. Hammacher A, Wijdenes J, Hilton DJ, Nicola NA, Simpson RJ, Layton JE . Ligand-specific utilization of the extracellular membrane-proximal region of the gp130-related signalling receptors Biochem J 2000 345: 25–32

    Article  CAS  Google Scholar 

  32. Bagby SP, O'Reilly MM, Kirk EA, Mitchell LH, Stenberg PE, Makler MT, Bakke AC . EGF is incomplete mitogen in porcine aortic smooth muscle cells: DNA synthesis without cell division Am J Phys 1992 262: C578–C588

    Article  CAS  Google Scholar 

  33. Schiemann WP, Nathanson NM . Box 3-independent signaling mechanisms are involved in leukemia inhibitory factor receptor alpha- and gp130-mediated stimulation of mitogen-activated protein kinase. Evidence for participation of multiple signaling pathways which converge at Ras J Biol Chem 1997 272: 16631–16636

    Article  CAS  Google Scholar 

  34. Chauhan D, Kharbanda SM, Ogata A, Urashima M, Frank D, Malik N, Kufe DW, Anderson KC . Oncostatin M induces association of Grb2 with Janus kinase JAK2 in multiple myeloma cells J Exp Med 1995 182: 1801–1806

    Article  CAS  Google Scholar 

  35. Zandstra PW, Lauffenburger DA, Eaves CJ . A ligand-receptor signaling threshold model of stem cell differentiation control: a biologically conserved mechanism applicable to hematopoiesis Blood 2000 96: 1215–1222

    CAS  PubMed  Google Scholar 

  36. Vereer PJ, Wouters FS, Reynolds ARH . BPI. Quantitative imaging of lateral ErbB1 receptor signal propagagtion in the plasma membrane Science 2000 290: 1567–1570

    Article  Google Scholar 

Download references

Acknowledgements

We thank Dr Anne Novak for critical reading of the manuscript. This work was supported by National Institutes of Health grant CA62242.

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

French, J., Tschumper, R. & Jelinek, D. Analysis of IL-6-mediated growth control of myeloma cells using a gp130 chimeric receptor approach. Leukemia 16, 1189–1196 (2002). https://doi.org/10.1038/sj.leu.2402516

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2402516

Keywords

This article is cited by

Search

Quick links