Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan.
J Orthop Res. 2012 Dec;30(12):1915-22. doi: 10.1002/jor.22157. Epub 2012 Jun 1.
MicroRNAs (miRNAs) are small RNAs of ∼22 base pairs that regulate gene expression. We harvested cartilage tissue from patients with polydactylism, anterior cruciate ligament injury, and osteoarthritis undergoing total knee arthroplasty and used microarrays to identify miRNAs whose expression is upregulated or downregulated with age. The results were assessed by real-time PCR and MTT assay in a mimic group, in which synthetic double-stranded RNA from the isolated miRNA was transfected to upregulate expression, and in an inhibitor group, in which the miRNA was bound specifically to downregulate expression. The expression of two miRNAs (miR-199a-3p and miR-193b) was upregulated with age and that of one miRNA (miR-320c) was downregulated with age. A real-time PCR assay showed that type 2 collagen, aggrecan, and SOX9 expression were downregulated in the miR-199a-3p mimic group but was upregulated in the inhibitor group. Similar results were observed for miR-193b. By contrast, ADAMTS5 expression was downregulated in the miR-320c mimic group and upregulated in the inhibitor group. Cell proliferative activity was upregulated significantly in the miR-193b inhibitor group compared with the control group. We believe that miR-199a-3p and miR-193b are involved in the senescence of chondrocytes, and miR-320c is involved in the juvenile properties of chondrocytes.
MicroRNAs (miRNAs) 是一种约 22 个碱基的小 RNA,可调节基因表达。我们从患有多指症、前交叉韧带损伤和骨关节炎的患者的软骨组织中采集样本,这些患者正在接受全膝关节置换术。我们使用微阵列来鉴定与年龄相关的表达上调或下调的 miRNA。通过实时 PCR 和 MTT 测定法在模拟组中评估结果,在该组中,分离的 miRNA 的合成双链 RNA 被转染以上调表达,在抑制剂组中,miRNA 被特异性结合以下调表达。两种 miRNA(miR-199a-3p 和 miR-193b)的表达随年龄的增长而上调,而一种 miRNA(miR-320c)的表达随年龄的增长而下调。实时 PCR 检测显示,miR-199a-3p 模拟组中的 2 型胶原、聚集蛋白聚糖和 SOX9 表达下调,而抑制剂组中的表达上调。miR-193b 也观察到类似的结果。相比之下,miR-320c 模拟组中的 ADAMTS5 表达下调,而抑制剂组中的表达上调。miR-193b 抑制剂组的细胞增殖活性与对照组相比显著上调。我们认为 miR-199a-3p 和 miR-193b 参与软骨细胞的衰老,miR-320c 参与软骨细胞的幼态性质。