Characterization of different osteoclast phenotypes in the progression of bone invasion by oral squamous cell carcinoma.
Characterization of different osteoclast phenotypes in the progression of bone invasion by oral squamous cell carcinoma.
复制标题
口腔鳞状细胞癌骨侵袭进展中不同破骨细胞表型的特征
DOI：
10.3892/or.2017.6166
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发表时间：
2018-03
期刊：
影响因子：
4.2
通讯作者：
Wang Z
中科院分区：
文献类型：
--
作者：
Quan J;Hou Y;Long W;Ye S;Wang Z
本研究的目的是描述口腔鳞状细胞癌（OSCC）骨浸润过程中破骨细胞的不同表型。将SCC-25人口腔鳞癌细胞接种于裸鼠颅骨中央，建立口腔鳞癌局部骨浸润模型，所有小鼠均出现典型的骨吸收区。抗酒石酸酸性磷酸酶（TRAP）染色显示肿瘤-骨界面有各种类型的巨大破骨细胞。从裸鼠体内分离骨髓细胞进行破骨细胞原代培养，但仅产生少量巨破骨细胞。此外，使用特殊的血液离心管来获得大量的外周血单核细胞（PBMC）。应用磁活化细胞分选技术（MACS），结合细胞因子集落刺激因子（CSF）和核因子-κB受体激活因子配体（RANKL），从人外周血单个核细胞（PBMC）中分离出破骨细胞。通过骨吸收测定进一步证实骨吸收。最后，将Transwell插入物用于SCC 25细胞和CD 14+单核细胞的间接细胞共培养。实时荧光定量PCR和免疫印迹法检测破骨细胞特异性标志物的表达。共培养3天和6天后，SCC 25细胞的条件培养基（CM）刺激破骨细胞标志物的表达，并通过TRAP和F-actin染色检测到额外的破骨细胞。在本研究中，在建立的骨侵袭动物模型中观察到不同的破骨细胞表型，并使用各种原代破骨细胞培养物进行证实。CM可促进破骨细胞标志物的表达，并诱导单核细胞分化为成熟的破骨细胞，其可吸收邻近的骨组织。
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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