miR-182 的过表达通过靶向 NAMPT 抑制黄韧带细胞的骨化。
Overexpression of miR-182 inhibits ossification of ligamentum flavum cells by targeting NAMPT.
机构信息
Department of Orthopaedics, The Affiliated Changzhou No. 2 People's Hospital with Nanjing Medical University, Changzhou, Jiangsu, China; Department of Spine Surgery, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
Department of Orthopaedics, The Affiliated Changzhou No. 2 People's Hospital with Nanjing Medical University, Changzhou, Jiangsu, China.
出版信息
Exp Cell Res. 2018 Jun 15;367(2):119-131. doi: 10.1016/j.yexcr.2018.03.008. Epub 2018 Mar 27.
Ossification of the ligamentum flavum (OLF) is a debilitating disease resulting from the development of ectopic bone formation, which leads to the compression of the spinal cord. Nicotinamide phosphoribosyltransferase (NAMPT) was found to be upregulated and microRNA-182 (miR-182) was downregulated in OLF tissue. We investigated the effects of NAMPT and miR-182 expression in OLF cells and the influence on proteins associated with osteogenic differentiation. MiR-182 overexpression inhibited NAMPT, RUNX2, OCN and OPN mRNA and protein expression in OLF cells. Alkaline phosphatase (ALP) assay and Alizarin red staining confirmed reduced levels of osteogenic differentiation and mineralized nodule formation. Knockdown of NAMPT and the NAMPT inhibitor FK866 also inhibits RUNX2, OCN and OPN mRNA expression and protein levels, whereas overexpression of NAMPT promotes the expression of RUNX2, OCN and OPN and the generation of bone nodules. Dual-luciferase reporter assays revealed that miR-182 directly targets NAMPT and downregulates its expression. Transfection of OLF cells with miR-182 downregulated NAMPT and suppressed the regulation of RUNX2, OCN, and OPN by NAMPT overexpression. Overall, these data demonstrate that miR-182 suppresses OLF by downregulating NAMPT.
黄韧带骨化（OLF）是一种使人衰弱的疾病，由异位骨形成引起，导致脊髓受压。研究发现，烟酰胺磷酸核糖转移酶（NAMPT）在 OLF 组织中上调，微小 RNA-182（miR-182）下调。我们研究了 NAMPT 和 miR-182 在 OLF 细胞中的表达及其对与成骨分化相关的蛋白质的影响。miR-182 过表达抑制 OLF 细胞中 NAMPT、RUNX2、OCN 和 OPN mRNA 和蛋白的表达。碱性磷酸酶（ALP）检测和茜素红染色证实成骨分化和矿化结节形成水平降低。NAMPT 敲低和 NAMPT 抑制剂 FK866 也抑制 RUNX2、OCN 和 OPN mRNA 表达和蛋白水平，而 NAMPT 过表达促进 RUNX2、OCN 和 OPN 的表达和骨结节的生成。双荧光素酶报告基因检测显示，miR-182 直接靶向 NAMPT 并下调其表达。转染 OLF 细胞 miR-182 下调 NAMPT 并抑制 NAMPT 过表达对 RUNX2、OCN 和 OPN 的调控。综上所述，这些数据表明 miR-182 通过下调 NAMPT 抑制 OLF。
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