Effects of methotrexate and salazosulfapyridine on protein profiles of exosomes derived from a human synovial sarcoma cell line of SW982

Purpose 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. Experimental design SW982 was treated with SASP and/or MTX under interleukin‐1β (IL‐1β)‐treated or nontreated condition...

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Published in:Proteomics. Clinical applications 2016-02, Vol.10 (2), p.164-171
Main Authors: Tsuno, Hirotaka, Suematsu, Naoya, Sato, Toshiyuki, Arito, Mitsumi, Matsui, Toshihiro, Iizuka, Nobuko, Omoteyama, Kazuki, Okamoto, Kazuki, Tohma, Shigeto, Kurokawa, Manae S., Kato, Tomohiro
Format: Article
Language:English
Subjects:
2D-DIGE
Antirheumatic Agents - pharmacology
Electrophoresis, Gel, Two-Dimensional
Exosome
Exosomes - drug effects
Exosomes - metabolism
Humans
Interleukin-1beta - metabolism
Methotrexate
Methotrexate - pharmacology
Neoplasm Proteins - analysis
Neoplasm Proteins - metabolism
Rheumatoid arthritis
Salazosulfapyridine
Sulfasalazine - pharmacology
Tumor Cells, Cultured
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Summary:Purpose 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. Experimental design 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 and clinical relevance 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.
ISSN:1862-8346
1862-8354
DOI:10.1002/prca.201500064