Characterization of different osteoclast phenotypes in the progression of bone invasion by oral squamous cell carcinoma

Quan,Jingjing; Hou,Yuluan; Long,Weiling; Ye,Shu; Wang,Zhiyuan

doi:10.3892/or.2017.6166

Characterization of different osteoclast phenotypes in the progression of bone invasion by oral squamous cell carcinoma

Authors:
- Jingjing Quan
- Yuluan Hou
- Weiling Long
- Shu Ye
- Zhiyuan Wang
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Affiliations: Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University and Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, Guangdong 510080, P.R. China, Affiliated High School-South China Normal University, Guangzhou, Guangdong 510630, P.R. China
Published online on: December 19, 2017 https://doi.org/10.3892/or.2017.6166
Pages: 1043-1051
Copyright: © Quan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to characterize different phenotypes of osteoclasts in the progression of bone invasion by oral squamous cell carcinoma (OSCC). A local bone invasion model of OSCC was established by injecting SCC25 human OSCC cells into the center of calvariae in nude mice, and all mice were found to have a typical bone resorption area. Staining for tartrate-resistant acid phosphatase (TRAP) revealed various types of giant osteoclasts in the tumour-bone interface. Bone marrow cells (BMCs) were isolated from the nude mice for primary osteoclast culture, but only a few giant osteoclasts were generated. Additionally, special blood centrifuge tubes were utilized to obtain large numbers of peripheral blood mononuclear cells (PBMCs). Using magnetic activated cell sorting (MACS) and the cytokines colony-stimulating factor (CSF) and receptor activator of nuclear factor-κb ligand (RANKL), we differentiated human osteoclasts from CD14+ monocytes of PBMCs. Bone resorption was further confirmed by a bone resorption assay. Finally, Transwell inserts were used for indirect cell co-culture of SCC25 cells and CD14+ monocytes. Expression of specific osteoclast markers was detected by real-time PCR and western blotting. After co-culture for 3 and 6 days, conditioned medium (CM) of SCC25 cells stimulated the expression of osteoclast markers, and additional osteoclasts were detected through staining of TRAP and F-actin. In the present study distinct osteoclast phenotypes were observed in the established bone invasion animal model, and were confirmed using various primary osteoclast cultures. CM of OSCC cells may promote the expression of osteoclast markers and induce the differentiation of monocytes to mature osteoclasts, which can resorb adjacent bone tissue.

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