Mutant monocyte chemoattractant protein-1 protein (7ND) inhibits osteoclast differentiation and reduces oral squamous carcinoma cell bone invasion.
Mutant monocyte chemoattractant protein-1 protein (7ND) inhibits osteoclast differentiation and reduces oral squamous carcinoma cell bone invasion.
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突变单核细胞趋化蛋白-1蛋白（7ND）抑制破骨细胞分化并减少口腔鳞癌细胞骨侵袭
DOI：
10.3892/ol.2018.8308
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发表时间：
2018-05
期刊：
影响因子：
2.9
通讯作者：
Quan J
中科院分区：
文献类型：
--
作者：
Luo S;Zhou C;Zhang J;Chen M;Li H;Zheng S;Quan J
7-氨基酸截短(7ND)蛋白是单核细胞趋化蛋白-1(MCP-1)的N端缺失突变体，具有显性负性抑制作用。7ND与野生型MCP-1形成异源二聚体，与MCP-1受体结合，抑制单核细胞趋化。本研究对7ND蛋白进行了克隆、表达和纯化。用四甲基偶氮唑盐比色法检测合成的7ND蛋白对口腔鳞癌SCC25细胞的增殖无明显影响。连续稀释的7ND蛋白用于单核细胞迁移和破骨细胞分化，抗酒石酸酸性磷酸酶染色显示，磁活化细胞分选显示，从分化簇14+(CD14+)单核细胞分化出的破骨细胞明显减少。免疫荧光证实了这些结果，在7ND处理的破骨细胞中观察到的F-肌动蛋白染色明显减少。此外，通过将SCC25细胞皮下注射到覆盖裸鼠颅骨的区域来检测骨侵袭。结果表明，含7ND蛋白的SCC25细胞的平均肿瘤体积与未处理的SCC25细胞的平均肿瘤体积相似。流式细胞仪分析显示，7ND处理组小鼠骨髓中CD14+细胞亚群较未处理组小鼠减少。显微计算机断层成像显示，注射SCC25细胞和7ND蛋白的颅骨骨吸收明显减少。综上所述，本研究的结果证明了7ND蛋白的潜在治疗价值。7ND MCP-1变异体不仅在体外具有抑制破骨细胞分化的功能，而且在体内还可以减缓口腔鳞癌细胞的骨侵袭进展。
The seven-amino acid truncated (7ND) protein is an N-terminal deletion mutant of monocyte chemoattractant protein-1 (MCP-1) and it functions as a dominant-negative inhibitor. 7ND and wild-type MCP-1 form a heterodimer, which binds to MCP-1 receptors and inhibits monocyte chemotaxis. In the present study, the 7ND protein was cloned, expressed and purified. An MTT assay revealed that the proliferation of oral squamous cell carcinoma (OSCC) SCC25 cells was not affected following 3 days of treatment with synthetic 7ND protein. Serial dilutions of the 7ND protein were tested for monocyte migration and osteoclast differentiation, and tartrate-resistant acid phosphatase staining demonstrated that significantly fewer osteoclasts were differentiated from cluster of differentiation 14+ (CD14+) monocytes using magnetic activated cell sorting. Immunofluorescence confirmed these results and significantly less F-actin staining was observed in 7ND-treated osteoclasts. Furthermore, bone invasion was examined by subcutaneously injecting SCC25 cells into the area overlaying the calvariae of nude mice. The results demonstrated that the average tumor volume of SCC25 cells with 7ND protein was similar to the average volume of tumors formed by untreated SCC25 cells. Flow cytometric analysis suggested that the CD14+ subpopulation in the bone marrow of 7ND-treated mice was reduced compared with that of untreated mice. Micro-computed tomography imaging revealed significantly less bone resorption in the calvariae injected with SCC25 cells plus the 7ND protein. Taken together, the results of the present study demonstrated the potential therapeutic value of the 7ND protein. The 7ND MCP-1 variant not only functions in vitro to inhibit osteoclast differentiation, but also reduces the progression of bone invasion by OSCC cells in vivo.
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发表时间：
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