Effect of CCN2 on FGF2-induced proliferation and MMP9 and MMP13 productions by chondrocytes.
Effect of CCN2 on FGF2-induced proliferation and MMP9 and MMP13 productions by chondrocytes.
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DOI：
10.1210/en.2011-0234
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发表时间：
2011-09
期刊：
影响因子：
4.8
通讯作者：
Takashi Nishida;S. Kubota;Eriko Aoyama;Danilo Janune;A. Maeda;M. Takigawa
中科院分区：
文献类型：
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作者：
Takashi Nishida;S. Kubota;Eriko Aoyama;Danilo Janune;A. Maeda;M. Takigawa
CCN2（又称结缔组织生长因子）通过其特有的四个模块与参与软骨内成骨的几种生长因子相互作用，并修饰这些生长因子的作用。目前，我们研究了CCN2是否与成纤维细胞生长因子2 （FGF2）相互作用。固相结合实验、免疫沉淀- western blot分析和表面等离子体共振（SPR）光谱分析表明，CCN2 （CT）的c端模块直接与FGF2结合，解离常数为5.5 nm。接下来，我们研究了CCN2和FGF2对培养软骨细胞增殖和基质金属蛋白酶(MMP)-9和-13产生的联合影响。FGF2不仅促进了MMP9和-13的增殖，还促进了MMP9和-13的产生，但FGF2与CT模块联合使用对MMP的产生和增殖均无促进作用。为了阐明其机制，我们研究了CCN2或其CT模块与FGF受体1的结合。结果，我们发现CCN2与FGF受体1结合的解离常数为362 nm，而CT模块则没有。此外，当我们在FGF2与CT模块联合刺激的软骨细胞HCS-2/8细胞中检测FGF信号传导时，与单独使用FGF2相比，ERK1/2、p38 MAPK和c-Jun n端激酶磷酸化水平降低。这些发现表明，CCN2可能通过与FGF2形成复合物作为FGF2功能的新型调节剂来调节软骨细胞的增殖和基质降解。
CCN2 (also known as connective tissue growth factor) interacts with several growth factors involved in endochondral ossification via its characteristic four modules and modifies the effect of such growth factors. Presently we investigated whether CCN2 interacts with fibroblast growth factor 2 (FGF2). Solid-phase binding assay, immunoprecipitation-Western blot analysis, and surface plasmon resonance (SPR) spectroscopy revealed that the C-terminal module of CCN2 (CT) directly bound to FGF2 with a dissociation constant of 5.5 nm. Next, we examined the combinational effects of CCN2 and FGF2 on the proliferation of and matrix metalloproteinase (MMP)-9 and -13 productions by cultured chondrocytes. FGF2 promoted not only the proliferation but also the production of MMP9 and -13, however, combined of FGF2 with CT module nullified the enhancement of both MMP productions and proliferation. To clarify the mechanism, we investigated the binding of CCN2 or its CT module to FGF receptor 1. As a result, we found that CCN2 bound to FGF receptor 1 with a dissociation constant of 362 nm, whereas the CT module did not. In addition, when we tested FGF signaling in chondrocytic HCS-2/8 cells stimulated by the combination of FGF2 with CT module, the level of ERK1/2, p38 MAPK, and c-Jun N-terminal kinase phosphorylation was decreased compared with that found with FGF2 alone. These findings suggest that CCN2 may regulate the proliferation and matrix degradation of chondrocytes by forming a complex with FGF2 as a novel modulator of FGF2 functions.