CD147通过MAPK途径促进RANKL介导的破骨细胞分化成熟的机制研究

Osteoclasts are large, multinucleated cells, which originate from the fusion of macrophages.They play a central role in bone resorption associated diseases and are thus important targets of anti-osteolytic therapy.Extracellular matrix metalloproteinase inducer (CD147) is a highly glycosylated transmembrane protein that is known to play a role in inflammation and tumor invasion and metastasis. CD147 is widely expressed in many tissues,and highly expressed in osteolytic associated diseases. But the relationship between CD147 and osteoclast is still unclear.In our previous work, we found that CD147 could promote the activation and lytic ability of osteoclast when co-incubate with RANKL(receptor activator of NF-kappaB ligand), and this effect can be significantly reduced by the CD147 blocking antibody in vitro，and ERK may play a key role in this reaction .Therefore, we assume that CD147 can promote the activation of osteoclast with the synergies of RANKL by up-regulating the MAPK pathway.In this study,we propose to use several osteoclast differentiation and maturation model，through gene transfection, silencing to observe the effect of CD147 on the RANKL related regulation factors and osteoclast differentiation related facors.The key focus is the change in the MAPK pathway under the effect of CD147.The work is to clarify the molecular mechanism of CD147 in promoting osteoclast differentiation and maturation, and provide the therapeutic targets for anti-osteolytic therapy.

破骨细胞（Osteoclast，OC）是各种溶骨性病变的基础，也是抗溶骨治疗的重要靶点。细胞外基质金属蛋白酶诱导剂（CD147）是广泛表达于人体多个组织的跨膜糖蛋白，主要参与炎症、肿瘤等的浸润和转移，在多种溶骨性病变中也有较高表达，而其与破骨细胞的关系尚不明确。前期工作中我们发现CD147可明显促进破骨细胞分化因子RANKL作用下的OC活化和溶骨能力，而阻断CD147能明显延缓OC的分化和成熟，进一步研究发现这一作用与MAPK通路中的胞外信号调节激酶ERK相关。因此我们认为：CD147通过协同RANKL可促进OC的分化成熟，MAPK通路在其中可能起着重要作用。本课题拟通过深入研究在CD147在协同RANKL作用中破骨细胞MAPK通路中的分子改变，同时观察CD147与RANKL通路中重要调控因子、OC分化成熟标志分子间的关系，明确CD147在促进OC分化成熟的作用及分子机制。

破骨细胞（Osteoclast，OC）是各种溶骨性病变的基础，也是抗溶骨治疗的重要靶点。细胞外基质金属蛋白酶诱导剂（CD147）是广泛表达于人体多个组织的跨膜糖蛋白，主要参与炎症、肿瘤等的浸润和转移，在多种溶骨性病变中也有较高表达，而其与破骨细胞的关系尚不明确。通过对课题进行深入研究发现：过表达肺癌A549细胞来源CD147促进肿瘤细胞的增殖、迁移及侵袭能力，干扰肺癌A549细胞来源CD147抑制肿瘤细胞的增殖、迁移及侵袭能力，表明肺癌A549细胞来源CD147促进肺癌肿瘤细胞的增殖、迁移及侵袭能力，其机制可能与β-catenin 的激活有关。同时发现：肺癌A549细胞来源CD147抑制成骨细胞分化，可能与敲除A549细胞CD147导致的DKK1表达下调存在一定关联；另外我们发现肺癌A549细胞条件培养基可明显促进破骨细胞的分化，而敲除CD147后该促进作用明显减弱，通过荧光定量PCR及ELISA检测发现敲除CD147后，A549细胞的IL-8 mRNA表达及条件培养基中IL-8蛋白表达均显著减少，而外源性IL-8重组蛋白可以促进破骨细胞的分化，利用IL-8中和抗体则可以阻断IL-8对破骨细胞的促分化作用，结果表明CD147通过影响肿瘤细胞IL-8的分泌促进破骨细胞的活化。我们还通过TRAP染色对破骨细胞在溶骨性骨转移病灶中进行定位，便于进一步分析CD147表达与破骨细胞引起的溶骨性破坏的相关性。相关实验结果已经发表在国外SCI期刊及国内核心期刊。通过本课题研究，我们初步阐明了肺癌细胞来源CD147对肺癌细胞生物学行为及其对破骨、成骨细胞分化的影响及相关机制，从而验证了CD147在肿瘤细胞的侵袭、转移以及肿瘤细胞导致骨损伤过程中的重要作用，为肿瘤细胞骨转移及肿瘤细胞导致骨损伤的预防提供了相关理论依据及治疗靶点。