IL-6-accelerated calcification by induction of ROR2 in human adipose tissue-derived mesenchymal stem cells is STAT3 dependent

Shunsuke Fukuyo, Kunihiro Yamaoka, Koshiro Sonomoto, Koichi Oshita, Yosuke Okada, Kazuyoshi Saito, Yasuhiro Yoshida, Tamotsu Kanazawa, Yasuhiro Minami, Yoshiya Tanaka

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51 Citations (Scopus)


Objective: The mechanisms of ectopic calcification in inflammatory diseases are poorly understood. We investigated the effects of inflammatory cytokines on the mechanisms of calcification in human adipose tissue-derived mesenchymal stem cells (hADSCs). Methods: The effects of inflammatory cytokines were evaluated using hADSCs cultured in osteoblast induction medium. mRNA expression was measured by real-time PCR and protein levels were measured by western blotting. Cell mineralization was evaluated by Alizarin Red S staining. Results: In hADSCs, administration of IL-6/soluble IL-6 receptor (sIL-6R), TNF or IL-1b accelerated calcification through enhanced expression of an osteoblast differentiation marker, runt-related transcription factor 2 (RUNX2). IL-6/sIL-6R had the greatest effect. The transcription of mRNA for receptor tyrosine kinase-like orphan receptor 2 (ROR2), involved in the non-canonical wingless-type (WNT) MMTV integration site pathway, was increased, while b-catenin expression, an essential factor in the canonical WNT signalling pathway for osteoblast differentiation, did not change. Suppression of signal transducer and activator of transcription 3 (STAT3), but not STAT1, by small interfering RNA (siRNA) exerted a strong inhibitory effect on RUNX2 and ROR2 expression, and inhibited accelerated calcification. Conclusion: IL-6/sIL-6R stimulation accelerated the ROR2/WNT5A pathway in hADSCs in a STAT3- dependent manner, resulting in augmented calcification. These results suggest that the mechanisms of ectopic calcification accelerated by IL-6 in hADSCs may be involved in chronic inflammatory tissues and that IL-6 inhibitors may be beneficial in the treatment of ectopic calcification in inflammatory diseases.

Original languageEnglish
Article numberket496
Pages (from-to)1282-1290
Number of pages9
JournalRheumatology (United Kingdom)
Issue number7
Publication statusPublished - 2014 Jul
Externally publishedYes


  • ADSCs
  • Ectopic calcification
  • IL-6
  • ROR2
  • STAT3
  • WNT5A

ASJC Scopus subject areas

  • Rheumatology
  • Pharmacology (medical)


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