Department of Orthopedics and Traumatology, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nangjing 210028, China; Department of Orthopedics and Traumatology, Jiangsu Province Academy of Traditional Chinese Medicine, Nangjing 210028, China.
Department of Traditional Chinese Medicine, Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, Nanjing Medical University Affiliated Cancer Hospital, Nangjing 210009, China.
Biomed Pharmacother. 2018 Jan;97:607-615. doi: 10.1016/j.biopha.2017.10.152. Epub 2017 Nov 6.
Osteoarthritis is a common cause of functional deterioration in older adults and is an immense burden on the aging population. The molecular mechanism underlying the regulation of chondrocyte requires further elucidation, particularly with respect to the role of microRNAs. The aim of this study was to identify and characterize the expression of miR-20 in normal and OA chondrocytes, and to determine its role in OA pathogenesis. MiR-20 expression in cartilage specimens was examined in 30 patients with knee osteoarthritis and 30 traumatic amputees. The effect of miR-20 on chondrocyte was also investigated in chondrocyte cell line. Transfection with miR-20 mimic or inhibitor was employed to investigate the effect of miR-20 on chondrocyte proliferation and autophagy. Cell proliferation activity was detected by MTT assay and clone formation, cell autophagy were evaluated by monodansylcadaverine staining and GFP-LC3 fluorescence microscopy. Western blotting and immunohistochemical were utilized to detect expressions of autophagy markers (LC3, Beclin1 and p62) and of relevant proteins in the PI3K/AKT/mTOR signaling pathway. The results demonstrated that miR-20 inhibit chondrocyte proliferation and autophagy by targeting ATG10 via PI3K/AKT/mTOR signaling pathway. Our data suggest that miR-20 has an important role in the pathogenesis of osteoarthritis and is a potential therapeutic target.
骨关节炎是老年人功能恶化的常见原因，也是老龄化人口的巨大负担。软骨细胞调控的分子机制尚需进一步阐明，特别是 microRNAs 的作用。本研究旨在鉴定和描述正常和 OA 软骨细胞中 miR-20 的表达，并确定其在 OA 发病机制中的作用。在 30 例膝关节骨关节炎患者和 30 例创伤性截肢患者的软骨标本中检测 miR-20 的表达。还在软骨细胞系中研究了 miR-20 对软骨细胞的影响。转染 miR-20 模拟物或抑制剂，以研究 miR-20 对软骨细胞增殖和自噬的影响。通过 MTT 检测和克隆形成检测细胞增殖活性，通过单丹磺酰戊二醛染色和 GFP-LC3 荧光显微镜评估细胞自噬。Western blot 和免疫组织化学用于检测自噬标志物（LC3、Beclin1 和 p62）和 PI3K/AKT/mTOR 信号通路中相关蛋白的表达。结果表明，miR-20 通过 PI3K/AKT/mTOR 信号通路靶向 ATG10 抑制软骨细胞增殖和自噬。我们的数据表明，miR-20 在骨关节炎发病机制中具有重要作用，是一个潜在的治疗靶点。
