Utilization of E-cadherin by monocytes from tumour cells plays key roles in the progression of bone invasion by oral squamous cell carcinoma
Utilization of E-cadherin by monocytes from tumour cells plays key roles in the progression of bone invasion by oral squamous cell carcinoma
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肿瘤细胞单核细胞对 E-钙粘蛋白的利用在口腔鳞状细胞癌骨侵袭进展中发挥关键作用
DOI：
10.3892/or.2017.5749
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发表时间：
2017-08-01
期刊：
影响因子：
4.2
通讯作者：
Zhang, Jingyuan
中科院分区：
文献类型：
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作者：
Quan, Jingjing;Du, Qian;Zhang, Jingyuan
最近有报道称，e -钙粘蛋白（E-cad）在破骨细胞发生的早期阶段表达，用中和抗体阻断E-cad可减少破骨细胞分化。由于我们之前的研究表明，E-cad蛋白在口腔鳞状细胞癌（OSCC）的骨侵袭中丢失，我们假设E-cad可能被单核细胞融合并分化为破骨细胞。本研究使用了两个研究模型来验证我们的假设。一方面，我们利用SCC25的OSCC细胞建立OSCC侵袭骨的动物模型，研究E-cad蛋白在体内是否消失；另一方面，我们采用SCC25与RAW 264.7细胞间接共培养模型，在转化生长因子- β 1 （tgf - β 1）的处理下，观察体外减少的E-cad蛋白是否被“劫持”。结果成功建立了骨侵犯的骨鳞癌动物模型。免疫组织化学（IHC）发现E-cad蛋白也有类似的变化，肿瘤细胞对其染色较弱。使用5 ng/ml tgf - β 1，我们证实SCC25细胞发生了人工上皮-间质转化（EMT），并改变了EMT标志物的表达和细胞形态。Real-time PCR结果显示，间接细胞共培养模型SCC25中E-cad mRNA表达减少，RAW 264.7中E-cad mRNA表达增加，免疫荧光（IF）观察到E-cad染色从SCC25向RAW 264.7明显切换。补充nf - κ B配体受体激活剂（RANKL）、抗酒石酸酸性磷酸酶（TRAP）和F-actin染色证实破骨细胞数量增加。综上所述，我们的研究发现了E-cad蛋白在OSCC骨侵袭过程中的转换。肿瘤细胞中E-cad的缺失可能被单核细胞利用分化为破骨细胞，从而进一步解释OSCC侵袭骨的潜在机制，这可能为未来的分子生物治疗提供线索。
E-cadherin (E-cad) is recently reported to be expressed in early stages of osteoclastogenesis, and blocking E-cad with neutralizing antibodies decreases osteoclast differentiation. Since our previous research demonstrates the loss of E-cad protein in the bone invasion by oral squamous cell carcinoma (OSCC), we hypothesize that E-cad may be utilized by monocytes to fuse and differentiate into osteoclasts. Two research models are used in the present study to explore our hypothesis. On one hand, we use OSCC cells of SCC25 to establish an animal model of bone invasion by OSCC, and investigate whether E-cad protein disappears in vivo; on the other hand, we use the indirect co-culture model of SCC25 and RAW 264.7 cells, with the treatment of transforming growth factor-beta 1 (TGF-beta 1), and observe whether the decreased E-cad protein is 'hijacked' in vitro. Results showed the animal model of OSCC with bone invasion was successfully established. Immunohistochemistry (IHC) found similar changes of E-cad protein, which was weakly stained by tumour cells. By using 5 ng/ml of TGF-beta 1, we confirmed the artificial epithelial-mesenchymal transition (EMT) of SCC25 cells, with changes of EMT marker expression and cell morphology. Real-time PCR showed E-cad mRNA decreased in SCC25 while increased in RAW 264.7 of the indirect cell co-culture model, and immunofluoresence (IF) observed the evident switch of E-cad staining from SCC25 to RAW 264.7. With the supplement of receptor activator of NF-kappa B ligand (RANKL), tartrate-resistant acid phosphatase (TRAP) and F-actin staining confirmed the increased number of osteoclasts. Taken together, our study found the switch of E-cad protein in the progression of bone invasion by OSCC. The loss of E-cad in tumour cells may be utilized by monocytes to differentiate into osteoclasts, thus further explaining the underlying mechanisms of bone invasion by OSCC, which may supply clues for future molecular biotherapies.