IL-6 and TNF-a Cooperate to Modulate the Cell Cycle of RA-Fibroblast-like Synoviocytes Via Cyclin Dependent Kinase Inhibitors

Kenta Kaneshiro¹, Kohsuke Yoshida¹, Ayako Nakai¹, Kohjin Suzuki¹, Koto Uchida¹, Teppei Hashimoto², Yoshiko Kawasaki³, Natsuko Nakagawa⁴, Koji Tateishi⁵, Nao Shibanuma⁶, Yoshitada Sakai⁷ and Akira Hashiramoto¹, ¹Department of Biophysics, Kobe University Graduate School of Health Sciences, Kobe, Japan, ²Department of Rheumatology, Kobe Kaisei Hospital, Kobe, Japan, ³The Center of Rheumatic Diseases, Department of Rheumatology, Kobe Kaisei Hospital, Kobe, Japan, ⁴Department of Orthopaedic Surgery, Konan-Kakogawa Hospital, Kakogawa, Japan, ⁵Orthpaedic Surgery, Kobe University Graduate School of Medicine, Kobe, Japan, ⁶Departmant of Orthopaedic Surgery, Kobe Kaisei Hospital, Kobe, Japan, ⁷Division of Rehabilitation Medicine, Kobe University Graduate School of Medicine, Kobe, Japan

Meeting: 2017 ACR/ARHP Annual Meeting

Date of first publication: September 18, 2017

Keywords: Fibroblasts, IL-6, rheumatoid arthritis (RA) and tumor necrosis factor (TNF)

Session Information

Date: Monday, November 6, 2017

Title: Cytokines, Mediators, Cell-Cell Adhesion, Cell Trafficking and Angiogenesis Poster II: Mesenchymal Cells Do React - But How?

Session Type: ACR Poster Session B

Session Time: 9:00AM-11:00AM

Background/Purpose:

IL-6 and TNF-α play an important role in the pathogenesis of RA, and the proliferation of RA-synoviocytes (FLS) is controlled by cell cycle regulators including Cyclins, Cyclin dependent kinases (CDK), CDK inhibitors (CKDIs) and retinoblastoma protein (RB). To reveal actions of proinflammatory cytokines on the cell cycle of RA-FLS, we examined the expressions of the cell cycle regulators and the cellular viability under stimulations of IL-6 and TNF-α.

Methods:

RA-FLS were cultured with or without IL-6/soluble IL-6 receptor (sIL-6R) (100ng/ml) or TNF-α (10ng/ml). The expressions of CDKIs (p16^INK4a, p21^Cip1, p27^Kip1) and Cyclin E1/2 mRNA were measured by Real-time PCR, CYCLIN D and CYCLIN E protein were measured by Western blot, the expression of CYCLIN D and the phosphorylation of RB were observed by immunofluorescence, and the cellular viabilities were measured by WST-8. In addition, siRNA/p27^Kip1 was introduced into RA-FLS to analyze the expression of Cyclin E1 mRNA by Real-time PCR, under stimulations with or without IL-6/sIL-6R or TNF-α.

Results:

IL-6/sIL-6R decreased the expression of p16^INK4a, whereas increased CYCLIN D and the phosphorylation of RB. TNF-α increased the expressions of p27^Kip1, Cyclin E1/2 mRNA, CYCLIN D, and the phosphorylation of RB. The expression of CYCLIN D and the phosphorylation of RB were synergistically increased by IL-6 and TNF-α. The expression of Cyclin E1 mRNA was decreased by siRNA/p27^Kip1.

Conclusion:

Results clearly indicate that IL-6 and TNF-α interact with each other in regulating the cell cycle of RA-FLS.

Disclosure: K. Kaneshiro, None; K. Yoshida, None; A. Nakai, None; K. Suzuki, None; K. Uchida, None; T. Hashimoto, None; Y. Kawasaki, None; N. Nakagawa, None; K. Tateishi, None; N. Shibanuma, None; Y. Sakai, None; A. Hashiramoto, None.

To cite this abstract in AMA style:

Kaneshiro K, Yoshida K, Nakai A, Suzuki K, Uchida K, Hashimoto T, Kawasaki Y, Nakagawa N, Tateishi K, Shibanuma N, Sakai Y, Hashiramoto A. IL-6 and TNF-a Cooperate to Modulate the Cell Cycle of RA-Fibroblast-like Synoviocytes Via Cyclin Dependent Kinase Inhibitors [abstract]. Arthritis Rheumatol. 2017; 69 (suppl 10). https://acrabstracts.org/abstract/il-6-and-tnf-a-cooperate-to-modulate-the-cell-cycle-of-ra-fibroblast-like-synoviocytes-via-cyclin-dependent-kinase-inhibitors/. Accessed .

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