Regulation of the IGFBP-5 and MMP-13 genes by the microRNAs miR-140 and miR-27a in human osteoarthritic chondrocytes.
Regulation of the IGFBP-5 and MMP-13 genes by the microRNAs miR-140 and miR-27a in human osteoarthritic chondrocytes.
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DOI：
10.1186/1471-2474-10-148
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发表时间：
2009-11-30
影响因子：
2.3
通讯作者：
Martel-Pelletier J
中科院分区：
文献类型：
--
作者：
Tardif G;Hum D;Pelletier JP;Duval N;Martel-Pelletier J
MMP-13和IGFBP-5是参与骨关节炎（OA）的重要因子。我们研究了miR-140和miR-27a这两个高度预测的microrna （mirna）是否在人OA软骨细胞中调节这两个基因。实时荧光定量PCR检测基因表达。通过将每种miRNA的前体（pre-miRNA）和抑制剂（anti-miRNA）瞬时转染人OA软骨细胞，评估每种miRNA对IGFBP-5和MMP-13表达/产生的影响。通过细胞因子和生长因子治疗OA软骨细胞，检测IGFBP-5、miR-140和miR-27a表达的调节。IGFBP-5在人软骨细胞中表达，在OA中表达水平显著降低（p < 0.04）。五种计算算法确定miR-140和miR-27a可能是MMP-13和IGFBP-5表达的调节因子。数据显示，这两种mirna均在软骨细胞中表达。与正常软骨细胞相比，OA中miR-140的表达显著降低（77%,p < 0.01），而miR-27a的表达仅轻微降低（23%）。转染pre-miR-140后，24小时IGFBP-5表达显著降低（p = 0.0002），转染anti-miR-140后，IGFBP-5表达显著升高（p = 0.05），而pre-miR-27a对MMP-13和IGFBP-5均无影响。抗mir -27a而非抗mir -140在72h后显著增加了MMP-13 （p < 0.05）和IGFBP-5 （p < 0.01）的表达。MMP-13和IGFBP-5蛋白的产生遵循与其表达谱相同的模式。这些数据表明，IGFBP-5是miR-140的直接靶点，而miR-27a可能间接下调MMP-13和IGFBP-5。这项研究首次显示了这些mirna在人OA软骨细胞中的调控作用。它们对参与OA病理生理的两个基因的影响增加了基因调控的复杂性，这可能为OA治疗策略开辟新的途径。
MMP-13 and IGFBP-5 are important factors involved in osteoarthritis (OA). We investigated whether two highly predicted microRNAs (miRNAs), miR-140 and miR-27a, regulate these two genes in human OA chondrocytes. Gene expression was determined by real-time PCR. The effect of each miRNA on IGFBP-5 and MMP-13 expression/production was evaluated by transiently transfecting their precursors (pre-miRNAs) and inhibitors (anti-miRNAs) into human OA chondrocytes. Modulation of IGFBP-5, miR-140 and miR-27a expression was determined upon treatment of OA chondrocytes with cytokines and growth factors. IGFBP-5 was expressed in human chondrocytes with its level significantly lower (p < 0.04) in OA. Five computational algorithms identified miR-140 and miR-27a as possible regulators of MMP-13 and IGFBP-5 expression. Data showed that both miRNAs were expressed in chondrocytes. There was a significant reduction (77%, p < 0.01) in miR-140 expression in OA compared to the normal chondrocytes, whereas miR-27a expression was only slightly decreased (23%). Transfection with pre-miR-140 significantly decreased (p = 0.0002) and with anti-miR-140 significantly increased (p = 0.05) IGFBP-5 expression at 24 hours, while pre-miR-27a did not affect either MMP-13 or IGFBP-5. Treatment with anti-miR-27a, but not with anti-miR-140, significantly increased the expression of both MMP-13 (p < 0.05) and IGFBP-5 (p < 0.01) after 72 hours of incubation. MMP-13 and IGFBP-5 protein production followed the same pattern as their expression profile. These data suggest that IGFBP-5 is a direct target of miR-140, whereas miR-27a down-regulates, likely indirectly, both MMP-13 and IGFBP-5. This study is the first to show the regulation of these miRNAs in human OA chondrocytes. Their effect on two genes involved in OA pathophysiology adds another level of complexity to gene regulation, which could open up novel avenues in OA therapeutic strategies.
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