Direct crosstalk between cancer and osteoblast lineage cells fuels metastatic growth in bone via auto-amplification of IL-6 and RANKL signaling pathways

J Bone Miner Res. 2014 Sep;29(9):1938-49. doi: 10.1002/jbmr.2231.

Abstract

The bone microenvironment and its modification by cancer and host cell interactions is a key driver of skeletal metastatic growth. Interleukin-6 (IL-6) stimulates receptor activator of NF-κB ligand (RANKL) expression in bone cells, and serum IL-6 levels are associated with poor clinical outcomes in cancer patients. We investigated the effects of RANKL on cancer cells and the role of tumor-derived IL-6 within the bone microenvironment. Using human breast cancer cell lines to induce tumors in the bone of immune-deficient mice, we first determined whether RANKL released by cells of the osteoblast lineage directly promotes IL-6 expression by cancer cells in vitro and in vivo. We then disrupted of IL-6 signaling in vivo either via knockdown of IL-6 in tumor cells or through treatment with specific anti-human or anti-mouse IL-6 receptor antibodies to investigate the tumor effect. Finally, we tested the effect of RANK knockdown in cancer cells on cancer growth. We demonstrate that osteoblast lineage-derived RANKL upregulates secretion of IL-6 by breast cancers in vivo and in vitro. IL-6, in turn, induces expression of RANK by cancer cells, which sensitizes the tumor to RANKL and significantly enhances cancer IL-6 release. Disruption in vivo of this auto-amplifying crosstalk by knockdown of IL-6 or RANK in cancer cells, or via treatment with anti-IL-6 receptor antibodies, significantly reduces tumor growth in bone but not in soft tissues. RANKL and IL-6 mediate direct paracrine-autocrine signaling between cells of the osteoblast lineage and cancer cells, significantly enhancing the growth of metastatic breast cancers within bone.

Keywords: BONE METASTASIS; BREAST CANCER; INTERLEUKIN-6; OSTEOBLAST; RANKL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Neoplasms / pathology
  • Bone Neoplasms / secondary*
  • Bone Resorption / pathology
  • Bone and Bones / drug effects
  • Bone and Bones / metabolism
  • Bone and Bones / pathology
  • Cell Line, Tumor
  • Cell Lineage* / drug effects
  • Coculture Techniques
  • Disease Progression
  • Gene Knockdown Techniques
  • Humans
  • Interleukin-6 / metabolism*
  • Mice
  • Neoplasm Metastasis
  • Neoplasms / blood
  • Neoplasms / pathology*
  • Osteoblasts / drug effects
  • Osteoblasts / pathology*
  • RANK Ligand / blood
  • RANK Ligand / metabolism*
  • Receptor Activator of Nuclear Factor-kappa B / metabolism
  • Receptors, Interleukin-6 / metabolism
  • Recombinant Proteins / pharmacology
  • Signal Transduction* / drug effects
  • Xenograft Model Antitumor Assays

Substances

  • Interleukin-6
  • RANK Ligand
  • Receptor Activator of Nuclear Factor-kappa B
  • Receptors, Interleukin-6
  • Recombinant Proteins
  • TNFRSF11A protein, human