Asporin Expression Is Highly Regulated in Human Chondrocytes
Asporin Expression Is Highly Regulated in Human Chondrocytes
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DOI：
10.2119/molmed.2011.00052
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发表时间：
2011-07-01
影响因子：
5.7
通讯作者：
Bauge, Catherine
中科院分区：
文献类型：
--
作者：
Duval, Elise;Bigot, Nicolas;Bauge, Catherine
最近报道了编码阿孢菌素的基因 D 重复序列的多态性与骨关节炎（最常见的关节疾病）之间的显着关联。本研究的目的是研究这种新的 I 类小富含亮氨酸蛋白聚糖 (SLRP) 在人关节软骨细胞中的表达。首先，我们通过蛋白质印迹和逆转录聚合酶链反应研究了细胞因子对阿孢菌素（ASPN）表达的调节。白介素 1 β 和肿瘤坏死因子 α 下调 ASPN，而转化生长因子 β 1（在无血清培养基中孵育时）上调 ASPN。与促炎细胞因子类似，细胞连续传代诱导的软骨细胞去分化伴随着阿孢菌素表达的减少，而通过三维培养的再分化则恢复了其表达。最后，我们发现转录因子Sp1在ASPN表达调控中的重要作用。 Sp1 异位表达增加了 ASPN mRNA 水平和启动子活性。此外，使用基因报告基因测定和电泳迁移率变动测定，我们发现Sp1通过位于ASPN基因转录起始位点上游-473和-140 bp之间的区域介导其作用。总之，本报告是第一个关于不同细胞因子在人关节软骨细胞中调节阿孢菌素表达的研究。我们的数据表明该基因的表达在软骨中受到精细调节，并表明 Sp1 的主要作用。 (C) 2011 年范斯坦医学研究所，www.feinsteininstitute.org
A significant association between a polymorphism in the D repeat of the gene encoding asporin and osteoarthritis, the most frequent of articular diseases, has been recently reported. The goal of the present study was to investigate the expression of this new class I small leucine-rich proteoglycan (SLRP) in human articular chondrocytes. First, we studied the modulation of asporin (ASPN) expression by cytokines by Western blot and reverse transcription-polymerase chain reaction. Interleukin-1 beta and tumor necrosis factor-alpha downregulated ASPN, whereas transforming growth factor-beta 1 (when incubated in a serum-free medium) upregulated it. Similarly to proinflammatory cytokines, chondrocyte dedifferentiation induced by a successive passages of cells was accompanied by a decreased asporin expression, whereas their redifferentiation by three-dimensional culture restored its expression. Finally, we found an important role of the transcription factor Sp1 in the regulation of ASPN expression. Sp1 ectopic expression increased ASPN mRNA level and promoter activity. In addition, using gene reporter assay and electrophoretic mobility shift assay, we showed that Sp1 mediated its effect through a region located between -473 and -140 bp upstream of the transcription start site in ASPN gene. In conclusion, this report is the first study on the regulation of asporin expression by different cytokines in human articular chondrocytes. Our data indicate that the expression of this gene is finely regulated in cartilage and suggest a major role of Sp1. (C) 2011 The Feinstein Institute for Medical Research, www.feinsteininstitute.org