Interleukin-6 and tumour necrosis factor-α cooperatively promote cell cycle regulators and proliferate rheumatoid arthritis fibroblast-like synovial cells
| Autor: | Kohjin Suzuki, Kenta Kaneshiro, Koji Tateishi, Akira Hashiramoto, Nao Shibanuma, Yoshiko Kawasaki, Yasuhiro Terashima, Yoshitada Sakai, Kohsuke Yoshida, Koto Uchida |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Cyclin E
Cyclin D Blotting Western Immunology Cell Enzyme-Linked Immunosorbent Assay Arthritis Rheumatoid 03 medical and health sciences 0302 clinical medicine Rheumatology Humans Immunology and Allergy Medicine 030212 general & internal medicine Viability assay neoplasms Cells Cultured 030203 arthritis & rheumatology biology Interleukin-6 Tumor Necrosis Factor-alpha business.industry Cell Cycle Retinoblastoma protein General Medicine Fibroblasts Cell cycle Synoviocytes Molecular biology Cyclin E1 medicine.anatomical_structure Gene Expression Regulation biology.protein RNA Cyclin-dependent kinase 6 biological phenomena cell phenomena and immunity business |
| Zdroj: | Scandinavian Journal of Rheumatology. 48(5):353-361 |
| ISSN: | 0300-9742 |
| Popis: | Objective: To elucidate the roles of interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) in cell cycle regulation and proliferation of rheumatoid arthritis fibroblast-like synovial cells (RA-FLSs). Methods: Under stimulation with IL-6/soluble interleukin-6 receptor (sIL-6R) and TNF-α, we examined the expression of cell cycle regulators [p16INK4a, p21Cip1, p27Kip1, cyclin-dependent kinase-4 (CDK4), CDK6, Cyclin D, Cyclin E, and retinoblastoma protein (pRB)] by quantitative polymerase chain reaction, Western blotting, and immunofluorescence staining. The expression of pRB, with or without 10% foetal bovine serum, was examined by Western blotting. DNA synthesis and cell viability were examined by the BrdU assay and WST-8 assay, respectively. After transfection with siRNA/p16INK4a, siRNA/p21Cip1, siRNA/p27Kip1, siRNA/CDK4, or siRNA/CDK6, RA-FLSs were successively stimulated with or without IL-6/sIL-6R or TNF-α to determine cell viability. Results: IL-6/sIL-6R significantly decreased the expression of p16INK4a, and increased p21Cip1, Cyclin E1, CYCLIN D, and pRB. TNF-α decreased the expression of CDK4, and significantly increased p27Kip1, CDK6, Cyclin E1/E2, CYCLIN D, CYCLIN E, pRB, and phosphorylated pRB (phospho-pRB). By immunofluorescence staining, CYCLIN D and phospho-pRB were simultaneously stained in the single cell. In serum-free culture, the expression of pRB was apparently decreased. DNA synthesis and cell viability were significantly increased by IL-6/sIL-6R and TNF-α. Silencing of CDK6 attenuated the cell viability induced by IL-6 and TNF-α. Conclusion: The results indicate that IL-6 and TNF-α interact with each other in regulating the cell cycle and accelerate the proliferation of RA-FLSs. |
| Databáze: | OpenAIRE |
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