SNAP-23 and VAMP-3 contribute to the release of IL-6 and TNFα from a human synovial sarcoma cell line

FEBS J. 2014 Feb;281(3):750-65. doi: 10.1111/febs.12620. Epub 2013 Dec 13.

Abstract

Fibroblast-like synoviocytes are important mediators of inflammatory joint damage in arthritis through the release of cytokines, but it is unknown whether their exocytosis from these particular cells is SNARE-dependent. Here, the complement of soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) in human synovial sarcoma cells (SW982) was examined with respect to the secretion of interleukin-6 (IL-6) and tumour necrosis factor α (TNFα), before and after knockdown of a synaptosome-associated protein of molecular mass 23 kDa (SNAP-23) or the vesicle-associated membrane protein 3 (VAMP-3). Wild-type SW982 cells expressed SNAP-23, VAMP-3, syntaxin isoforms 2-4 and synaptic vesicle protein 2C (SV2C). These cells showed Ca²⁺-dependent secretion of IL-6 and TNFα when stimulated by interleukin-1β (IL-1β) or in combination with K⁺ depolarization. Specific knockdown of SNAP-23 or VAMP-3 decreased the exocytosis of IL-6 and TNFα; the reduced expression of SNAP-23 caused accumulation of SV2 in the peri-nuclear area. A monoclonal antibody specific for VAMP-3 precipitated SNAP-23 and syntaxin-2 (and syntaxin-3 to a lesser extent). The formation of SDS-resistant complexes by SNAP-23 and VAMP-3 was reduced upon knockdown of SNAP-23. Although the syntaxin isoforms 2, 3 and 4 are expressed in SW982 cells, knockdown of each did not affect the release of cytokines. Collectively, these results show that SNAP-23 and VAMP-3 participate in IL-1β-induced Ca²⁺-dependent release of IL-6 and TNFα from SW982 cells.

Keywords: IL-6; SNAREs; SW982; TNFα; arthritis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Inflammatory Agents / pharmacology
  • Anti-Inflammatory Agents / therapeutic use
  • Arthritis / drug therapy
  • Arthritis / immunology
  • Arthritis / metabolism
  • Calcium Signaling / drug effects
  • Cell Line, Tumor
  • Exocytosis* / drug effects
  • Humans
  • Interleukin-1beta / metabolism
  • Interleukin-6 / metabolism*
  • Membrane Glycoproteins / antagonists & inhibitors
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / metabolism
  • Molecular Targeted Therapy
  • Nerve Tissue Proteins / antagonists & inhibitors
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Protein Isoforms / antagonists & inhibitors
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Protein Transport / drug effects
  • Qa-SNARE Proteins / antagonists & inhibitors
  • Qa-SNARE Proteins / genetics
  • Qa-SNARE Proteins / metabolism
  • Qb-SNARE Proteins / antagonists & inhibitors
  • Qb-SNARE Proteins / genetics
  • Qb-SNARE Proteins / metabolism*
  • Qc-SNARE Proteins / antagonists & inhibitors
  • Qc-SNARE Proteins / genetics
  • Qc-SNARE Proteins / metabolism*
  • RNA Interference
  • RNA, Small Interfering
  • Receptors, Interleukin-1 / metabolism
  • Synovial Membrane / drug effects
  • Synovial Membrane / immunology
  • Synovial Membrane / metabolism*
  • Tumor Necrosis Factor-alpha / metabolism*
  • Vesicle-Associated Membrane Protein 3 / antagonists & inhibitors
  • Vesicle-Associated Membrane Protein 3 / genetics
  • Vesicle-Associated Membrane Protein 3 / metabolism*

Substances

  • Anti-Inflammatory Agents
  • IL6 protein, human
  • Interleukin-1beta
  • Interleukin-6
  • Membrane Glycoproteins
  • Nerve Tissue Proteins
  • Protein Isoforms
  • Qa-SNARE Proteins
  • Qb-SNARE Proteins
  • Qc-SNARE Proteins
  • RNA, Small Interfering
  • Receptors, Interleukin-1
  • SNAP23 protein, human
  • SV2C protein, human
  • TNF protein, human
  • Tumor Necrosis Factor-alpha
  • VAMP3 protein, human
  • Vesicle-Associated Membrane Protein 3