Interleukin‑1 induces receptor activator of nuclear factor‑κB ligand‑independent osteoclast differentiation in RAW264.7 cells

Liao,Rongdong; Feng,Zhuoxi; Li,Wei; Liu,Rubing; Xu,Xinrou; Yao,Shun; Tian,Jing

doi:10.3892/etm.2021.10072

Interleukin‑1 induces receptor activator of nuclear factor‑κB ligand‑independent osteoclast differentiation in RAW264.7 cells

Authors:
- Rongdong Liao
- Zhuoxi Feng
- Wei Li
- Rubing Liu
- Xinrou Xu
- Shun Yao
- Jing Tian
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Affiliations: Department of Orthopedics, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510280, P.R. China, The Second Clinical Medical School, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510280, P.R. China, Department of Clinical Skills Training Center, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510280, P.R. China
Published online on: April 16, 2021 https://doi.org/10.3892/etm.2021.10072
Article Number: 640
Copyright: © Liao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Interleukin‑1 (IL‑1) is a pro‑inflammatory cytokine which induces bone destruction in various diseases, such as osteoporosis and rheumatoid arthritis. RAW264.7 cells are frequently used in studies as osteoclast precursors, however it remains unclear whether IL‑1 can induce osteoclast differentiation from RAW264.7 cells without the stimulation of receptor activator of nuclear factor‑κB ligand (RANKL). Hence, the present study aimed to investigate the effects of IL‑1 on the formation of osteoclasts from RAW264.7 cells. The cell viability was determined via the Cell Counting Kit‑8 (CCK‑8) assay. Protein and gene expression were measured by western blotting and reverse transcription‑quantitative PCR, respectively. Tartrate‑resistant acid phosphatase (TRAP) staining and the resorption pit assay were performed to determine the formation and activity of osteoclasts. A significantly increased quantity of osteoclasts were found in the IL‑1 group compared with the control group, and also in the RANKL+IL‑1 group compared with the RANKL group. In addition IL‑1 significantly increased both the protein and mRNA expression of specific genes associated with osteoclastogenesis, including nuclear factor of activated T cells cytoplasmic 1, matrix metalloprotein‑9, cathepsin K and TRAP. The findings of the present study suggested that IL‑1 can induce osteoclast differentiation and upregulate the quantity of osteoclasts differentiated from RAW264.7 cells. These results may lay a foundation for further study of diseases involving inflammation‑associated bone loss. The combined blockade of IL‑1 and RANKL may be effective for the prevention of inflammatory bone loss.

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