MicroRNA-125b regulates the expression of aggrecanase-1 (ADAMTS-4) in human osteoarthritic chondrocytes.
MicroRNA-125b regulates the expression of aggrecanase-1 (ADAMTS-4) in human osteoarthritic chondrocytes.
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MicroRNA-125b 调节人骨关节炎软骨细胞中聚集蛋白聚糖酶 1 (ADAMTS-4) 的表达。
DOI：
10.1186/ar4164
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发表时间：
2013-02-13
影响因子：
4.9
通讯作者：
Ishiguro N
中科院分区：
文献类型：
--
作者：
Matsukawa T;Sakai T;Yonezawa T;Hiraiwa H;Hamada T;Nakashima M;Ono Y;Ishizuka S;Nakahara H;Lotz MK;Asahara H;Ishiguro N
聚集蛋白聚糖酶-1（ADAMTS-4）的表达增加已成为骨关节炎（OA）和其他关节疾病的重要因素。本研究旨在确定ADAMTS-4在人软骨细胞中的表达是否受miRNA的调控。使用生物信息学鉴定miRNA靶标。从膝关节软骨中分离软骨细胞，并用白细胞介素-1 β（IL-1β）处理。使用TaqMan测定法定量基因表达，并通过免疫印迹法测定蛋白质产生。荧光素酶报告基因分析用于验证miRNA与靶信使RNA（mRNA）之间的相互作用。计算机模拟分析预测了ADAMTS-4上miR-125 b的推定靶序列。miR-125 b在正常和OA软骨细胞中均有表达，OA软骨细胞中的表达显著低于正常软骨细胞。此外，IL-1β诱导的ADAMTS-4上调被人OA软骨细胞中miR-125 b的过表达所抑制。在荧光素酶报告基因测定中，ADAMTS-4 3 'UTR中推定的miR-125 b结合位点的突变消除了miR 125的抑制作用。我们的研究结果表明，miR-125 b在调节人软骨细胞中ADAMTS-4的表达中起重要作用，这将miR-125 b确定为OA的新治疗靶点。
Increased expression of aggrecanase-1 (ADAMTS-4) has emerged as an important factor in osteoarthritis (OA) and other joint diseases. This study aimed to determine whether the expression of ADAMTS-4 in human chondrocytes is regulated by miRNA. MiRNA targets were identified using bioinformatics. Chondrocytes were isolated from knee cartilage and treated with interleukin-1 beta (IL-1β). Gene expression was quantified using TaqMan assays and protein production was determined by immunoblotting. Luciferase reporter assay was used to verify interaction between miRNA and target messenger RNA (mRNA). In silico analysis predicted putative target sequence of miR-125b on ADAMTS-4. MiR-125b was expressed in both normal and OA chondrocytes, with significantly lower expression in OA chondrocytes than in normal chondrocytes. Furthermore, IL-1β-induced upregulation of ADAMTS-4 was suppressed by overexpression of miR-125b in human OA chondrocytes. In the luciferase reporter assay, mutation of the putative miR-125b binding site in the ADAMTS-4 3'UTR abrogated the suppressive effect of miR125. Our results indicate that miR-125b plays an important role in regulating the expression of ADAMTS-4 in human chondrocytes and this identifies miR-125b as a novel therapeutic target in OA.
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影响因子：
56.9
作者：
Lagos-Quintana, M;Rauhut, R;Tuschl, T
通讯作者：
Tuschl, T
影响因子：
2.2
作者：
Naito, Satoko;Shiomi, Takayuki;Okada, Yasunori
通讯作者：
Okada, Yasunori
影响因子：
5
作者：
Goldring, M B
通讯作者：
Goldring, M B
DOI：
10.1016/0167-4889(90)90145-4
发表时间：
1990-05-22
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
LEFEBVRE, V;PEETERSJORIS, C;VAES, G
通讯作者：
VAES, G
影响因子：
7
作者：
Tortorella, MD;Malfait, AM;Arner, E
通讯作者：
Arner, E