RECK Is Up-Regulated and Involved in Chondrocyte Cloning in Human Osteoarthritic Cartilage
RECK Is Up-Regulated and Involved in Chondrocyte Cloning in Human Osteoarthritic Cartilage
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DOI：
10.2353/ajpath.2010.091003
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发表时间：
2010-06-01
影响因子：
6
通讯作者：
Okada, Yasunori
中科院分区：
文献类型：
--
作者：
Kimura, Tokuhiro;Okada, Aiko;Okada, Yasunori
具有Kazal基序的逆转诱导富含半胱氨酸蛋白(RECK)是一种膜锚定的基质金属蛋白酶调节因子，但其在软骨中的功能尚不完全清楚。本研究的目的是研究RECK在人骨关节炎(OA)软骨中的表达和功能。定量RT-PCR结果显示，RECK在骨性关节炎软骨中的表达水平明显高于正常软骨。免疫组织化学分析显示，RECK定位于OA软骨中的软骨细胞，其免疫反应性与软骨细胞的Mankin评分、细胞克隆和增殖程度直接相关。在培养的骨性关节炎软骨细胞中，RECK通过糖基磷脂酰肌醇锚定在细胞表面表达。胰岛素样生长因子-1刺激其表达，白介素1和肿瘤坏死因子-a抑制其表达。小干扰RNA下调RECK的表达显示软骨细胞的铺展减少，局部粘连减少。在单层损伤实验中，RECK的下调可促进软骨细胞的迁移，而RECK的过表达则以一种依赖于基质金属蛋白酶活性的方式抑制软骨细胞的迁移。另一方面，RECK的沉默抑制了软骨细胞的增殖，这与粘着斑激酶和细胞外信号调节激酶的磷酸化减少有关，而RECK的过表达则促进了软骨细胞的增殖。这些数据首次证明RECK在人骨关节炎软骨中表达上调，并提示RECK可能通过抑制和促进软骨细胞的迁移和增殖而在软骨细胞克隆中发挥作用。(Am J Pathol2010,17628582867；DOI：10.2353/ajpath.2010.091003)
Reversion-inducing cysteine-rich protein with Kazal motifs (RECK) is a membrane-anchored matrix metalloproteinase regulator, but its functions in cartilage are not fully understood. The aim of the present study was to examine the expression and functions of RECK in human osteoarthritic (OA) cartilage. Quantitative RT-PCR indicated that the expression level of RECK is significantly higher in OA cartilage than in normal cartilage. By immunohistochemical analysis, RECK was localized to chondrocytes in OA cartilage, and the immunoreactivity directly correlated with the Mankin score and degree of chondrocyte cloning and proliferation. In cultured OA chondrocytes, RECK was expressed on the cell surface by glycosylphosphatidylinositol anchoring. The expression was stimulated by insulin-like growth factor-1 and suppressed by interleukin-1 and tumor necrosis factor-a. Down-regulation of RECK by small interfering RNA showed reduced spreading and smaller focal adhesions in the chondrocytes. Chondrocyte migration in a monolayer wounding assay was increased by down-regulation of RECK and inhibited by RECK overexpression in an matrix metalloproteinase activity-dependent manner. On the other hand, chondrocyte proliferation was suppressed by RECK silencing, and this was associated with reduced phosphorylation of focal adhesion kinase and extracellular signal-regulated kinase, whereas the proliferation was enhanced by RECK overexpression. These data are the first to demonstrate that RECK is up-regulated in human OA cartilage and suggest that RECK plays a role in chondrocyte cloning probably through suppression and promotion of chondrocyte migration and proliferation, respectively. (Am J Pathol 2010, 176-2858-2867; DOI: 10.2353/ajpath.2010.091003)