Department of Orthopedics, Changzheng Hospital, Second Military Medical University, Shanghai, China.
State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
Acta Biochim Biophys Sin (Shanghai). 2019 Jun 20;51(6):571-579. doi: 10.1093/abbs/gmz036.
The functional changes of nucleus pulposus (NP) cells are considered to be the initiating factors of intervertebral disc degeneration (IDD), and the differentially expressed circRNAs in NP cells may play an important role in the process of IDD. To identify circular RNAs (circRNAs) associated with human IDD, we isolated the NP cells from human degenerated and non-degenerated intervertebral disc and identified NP cells by microscopy and cell proliferation. CircRNA microarray expression profiles were obtained from NP cells of degenerated and non-degenerated intervertebral disc and further validated by quantitative reverse transcription PCR (qRT-PCR). The expression data were analyzed by bioinformatics. Microarray analysis identified 7294 circRNAs differentially expressed in degenerated human IDD NP cells. Among them, 3724 circRNAs were up-regulated and 3570 circRNAs were down-regulated by more than 2 folds. After validating by qRT-PCR, we predicted the possible miRNAs of the top dysregulated circRNAs using TargetScan, and miRanda. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the most modulated circRNAs regulate the viability, degradation, apoptosis and oxidative stress in NP cells, and the possible mechanism underlying IDD was discussed. These results revealed that circRNAs may play a role in IDD and might be a promising candidate molecular target for gene therapy.
核髓细胞（NP）的功能变化被认为是椎间盘退行性变（IDD）的起始因素，而 NP 细胞中差异表达的 circRNAs 可能在 IDD 过程中发挥重要作用。为了鉴定与人类 IDD 相关的环状 RNA（circRNA），我们从人退变和非退变椎间盘分离 NP 细胞，并通过显微镜和细胞增殖鉴定 NP 细胞。从退变和非退变椎间盘的 NP 细胞中获得 circRNA 微阵列表达谱，并通过定量逆转录 PCR（qRT-PCR）进一步验证。通过生物信息学分析表达数据。微阵列分析鉴定出 7294 个在退变人类 IDD NP 细胞中差异表达的 circRNAs。其中，3724 个 circRNAs 上调，3570 个 circRNAs 下调 2 倍以上。通过 qRT-PCR 验证后，我们使用 TargetScan 和 miRanda 预测了 top 失调 circRNAs 的可能 miRNA。此外，京都基因与基因组百科全书（KEGG）通路分析表明，最调节的 circRNAs 调节 NP 细胞的活力、降解、凋亡和氧化应激，讨论了 IDD 的可能机制。这些结果表明 circRNAs 可能在 IDD 中发挥作用，并且可能是基因治疗的有前途的候选分子靶标。
