MicroRNA-9 regulates the development of knee osteoarthritis through the NF-kappaB1 pathway in chondrocytes.
MicroRNA-9 regulates the development of knee osteoarthritis through the NF-kappaB1 pathway in chondrocytes.
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DOI：
10.1097/md.0000000000004315
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发表时间：
2016-09
期刊：
影响因子：
1.6
通讯作者：
Yang J
中科院分区：
文献类型：
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作者：
Gu R;Liu N;Luo S;Huang W;Zha Z;Yang J
已有研究表明，microRNA-9 （miR-9）与膝关节骨关节炎（OA）的发展有关。本研究旨在探讨miR-9靶向核因子κ b1 （NF-κB1）与膝关节OA软骨细胞增殖和凋亡的关系机制。取25例膝关节OA患者和10例外伤性截肢患者的软骨标本，建立OA大鼠模型15只，无膝关节OA病变大鼠15只。采用实时荧光定量PCR检测MiR-9在膝关节OA软骨和正常软骨中的表达。同时检测两组相关基因NF-κB1、IL-6、MMP-13的表达。采用双荧光素酶报告基因法检测miR-9对NF-κB1 3'UTR荧光素酶活性的影响。分别用miR-9模拟物、miR-9抑制剂和NF-κB1 siRNA转染膝关节OA软骨细胞，分别通过MTT法和流式细胞术检测细胞增殖和凋亡的变化。Western blotting检测NF-κB1、白细胞介素-6 （IL-6）、基质金属蛋白酶-13 （MMP-13）的表达。人OA样本和大鼠OA模型结果显示，与正常软骨组织相比，miR-9在膝关节OA软骨组织中明显下调（P < 0.01）。膝关节OA软骨组织中NF-κB1、IL-6、MMP-13的表达显著高于正常软骨组织（P < 0.01）。双荧光素酶报告基因检测显示，miR-9可结合NF-κB1的3'UTR，显著抑制荧光素酶活性37% （P < 0.01）。上调miR-9或下调NF-κB1可促进细胞增殖，抑制细胞凋亡。总之，下调的miR-9可以通过直接结合NF-kB1促进膝关节OA软骨细胞的增殖和抗凋亡，这意味着刺激miR-9的表达可能有助于治疗膝关节OA。
Supplemental Digital Content is available in the text It has been suggested that microRNA-9 (miR-9) is associated with the development of knee osteoarthritis (OA). This study was aimed to investigate the association between the mechanism of miR-9 targeting nuclear factor kappa-B1 (NF-κB1) and the proliferation and apoptosis of knee OA chondrocytes. Cartilage samples were collected from 25 patients with knee OA and 10 traumatic amputees, and another 15 OA rat models, together with 15 rats without knee OA lesions were also established. MiR-9 expressions in both knee OA cartilage and normal cartilage samples were detected using quantitative real-time PCR. The expressions of related genes (NF-κB1, IL-6, and MMP-13) in the two groups were also detected. Dual luciferase reporter gene assay was employed to examine the effect of miR-9 on the luciferase activity of NF-κB1 3′UTR. Knee OA chondrocytes were transfected with miR-9 mimics, miR-9 inhibitor, and NF-κB1 siRNA, respectively, and changes in cellular proliferation and apoptosis were detected via MTT assay and flow cytometric analysis, respectively. Western blotting assay was used to detect the expressions of NF-κB1, interleukin-6 (IL-6), and matrix metalloproteinase-13 (MMP-13). According to results from human OA samples and rat OA models, miR-9 was significantly downregulated in knee OA cartilage tissues compared with normal cartilage tissues (P < 0.01). The expressions of NF-κB1, IL-6, and MMP-13 in knee OA cartilage tissues were significantly higher than those in normal cartilage tissues (P < 0.01). Dual luciferase reporter gene assay showed that miR-9 could bind to the 3′UTR of NF-κB1 and significantly inhibit the luciferase activity by 37% (P < 0.01). Upregulation of miR-9 or downregulation of NF-κB1 could promote cell proliferation and suppress cell apoptosis. Conclusively, downregulated miR-9 can facilitate proliferation and antiapoptosis of knee OA chondrocytes by directly binding to NF-kB1, implying that stimulating miR-9 expressions might assist in treatment of knee OA.