title: Effects of methotrexate and salazosulfapyridine on protein profiles of exosomes derived from a human synovial sarcoma cell line of SW982.
abstract: OBJECTIVE
  To elucidate effects of salazosulfapyridine (SASP) and methotrexate (MTX), major anti-rheumatic drugs, on exosomes derived from SW982 of a human synovial sarcoma cell line.
  
  
  METHODS
  SW982 was treated with SASP and/or MTX under interleukin-1β (IL-1β)-treated or nontreated conditions. Exosomes were isolated from the culture media, and exosomal proteome was analyzed by 2D-DIGE. Protein spots whose intensity was significantly altered by the above treatments were identified by MS.
  
  
  RESULTS
  Two hundred ninety-four protein spots were detected in the exosome preparations by 2D-DIGE. Compared to the nontreated cells, SASP-, MTX-, and (SASP + MTX)-treated cells displayed 8, 10, and 21 exosomal protein spots with more than ±2.0-fold intensity differences (p < 0.05), respectively. Similarly, the IL-1β-treated cells displayed 58 exosomal protein spots with more than ±1.5-fold intensity differences (p < 0.05). In about half of the 58 spots, the IL-1β-induced intensity changes were suppressed by simultaneous addition of SASP and/or MTX. Most of the identified proteins were immunity- or anti-oxidation-related proteins.
  
  
  CONCLUSIONS
  The SASP and/or MTX treatments altered the protein profiles of exosomes and suppressed the effects of IL-1β on the exosomal proteome. Exosomes may play roles in the actions of these anti-rheumatic drugs.
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Sugao, Miyamae, Kawasaki, Kanagawa, Japan.|Department of Rheumatology, Sagamihara National Hospital, National Hospital Organization, Sakuradai, Minami, Sagamihara, Kanagawa, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Tsuno
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Sugao, Miyamae, Kawasaki, Kanagawa, Japan.
authors.forename: Naoya
authors.initials: N
authors.lastname: Suematsu
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Sugao, Miyamae, Kawasaki, Kanagawa, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Sato
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Sugao, Miyamae, Kawasaki, Kanagawa, Japan.
authors.forename: Mitsumi
authors.initials: M
authors.lastname: Arito
authors.affiliation: Department of Rheumatology, Sagamihara National Hospital, National Hospital Organization, Sakuradai, Minami, Sagamihara, Kanagawa, Japan.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Matsui
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Sugao, Miyamae, Kawasaki, Kanagawa, Japan.
authors.forename: Nobuko
authors.initials: N
authors.lastname: Iizuka
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Sugao, Miyamae, Kawasaki, Kanagawa, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Omoteyama
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Sugao, Miyamae, Kawasaki, Kanagawa, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Okamoto
authors.affiliation: Department of Rheumatology, Clinical Research Center for Allergy and Rheumatology, Sagamihara National Hospital, National Hospital Organization, Sakuradai, Minami, Sagamihara, Kanagawa, Japan.
authors.forename: Shigeto
authors.initials: S
authors.lastname: Tohma
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Sugao, Miyamae, Kawasaki, Kanagawa, Japan.
authors.forename: Manae S
authors.initials: MS
authors.lastname: Kurokawa
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Sugao, Miyamae, Kawasaki, Kanagawa, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Kato
mesh_terms: D018501:Antirheumatic Agents / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D055354:Exosomes / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism; D008727:Methotrexate / Q000494:pharmacology*; D009363:Neoplasm Proteins / Q000032:analysis / Q000378:metabolism*; D012460:Sulfasalazine / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D018501:Antirheumatic Agents; D053583:Interleukin-1beta; D009363:Neoplasm Proteins; D012460:Sulfasalazine; D008727:Methotrexate
keywords: 2D-DIGE; Exosome; Methotrexate; Rheumatoid arthritis; Salazosulfapyridine