Effects of Ca2+/calmodulin‑dependent protein kinase pathway inhibitor KN93 on osteoclastogenesis

Fu,Yingxiao; Niu,Dequn; Su,Wenfang; Yang,Qingling; Wang,Wenrui; Tang,Baoding; Li,Zhongwen; Zhang,Ding; Mao,Yingji; Li,Chuang; Li,Xue; Ye,Shihao; Su,Xu; Xu,Fanyuan; Sun,Xuemin; Chen,Changjie

doi:10.3892/ijmm.2018.3793

Effects of Ca2+/calmodulin‑dependent protein kinase pathway inhibitor KN93 on osteoclastogenesis

Authors:
- Yingxiao Fu
- Dequn Niu
- Wenfang Su
- Qingling Yang
- Wenrui Wang
- Baoding Tang
- Zhongwen Li
- Ding Zhang
- Yingji Mao
- Chuang Li
- Xue Li
- Shihao Ye
- Xu Su
- Fanyuan Xu
- Xuemin Sun
- Changjie Chen
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Affiliations: Department of Bioscience, Bengbu Medical College, Bengbu, Anhui 233000, P.R. China, Department of Gynecology and Obstetrics, The Second Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233000, P.R. China, Department of Medical Laboratory, Bengbu Medical College, Bengbu, Anhui 233000, P.R. China, Department of Clinical Medicine, Bengbu Medical College, Bengbu, Anhui 233000, P.R. China
Published online on: July 25, 2018 https://doi.org/10.3892/ijmm.2018.3793
Pages: 2294-2302

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Abstract

The aim of the present study was to determine the effects of the Ca2+/calmodulin‑dependent protein kinase pathway inhibitor KN93 on osteoclastogenesis. RAW264.7 cells were incubated with macrophage colony‑stimulating factor (M‑CSF) + receptor activator of nuclear factor kappa‑light‑chain‑enhancer of activated B cells ligand (RANKL) to stimulate osteoclastogenesis and then treated with 10 µM KN93. The methods included tartrate‑resistant acid phosphatase (TRAP) staining, bone resorption activity assays, filamentous (F)‑actin staining, determination of intracellular calcium ([Ca2+]i) levels, monitoring of osteoclast‑specific gene expression levels and measurement of key transcription factors protein levels. The results suggested that KN93 inhibited the formation of TRAP‑positive multinucleated cells, shaping of F‑actin rings and resorption activity of the cells. In addition, KN93 decreased the concentration of [Ca2+]i, expression levels of osteoclast specific genes and protein levels of critical transcription factors in the M‑CSF + RANKL‑induced osteoclast model. In summary, KN93 may directly affect the differentiation and activation of osteoclasts, potentially through the Ca2+/calmodulin‑dependent protein kinase signaling pathway.

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