Trauma Centre, Department of Trauma and Orthopedics, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai 201620, P.R. China.
Sino Euro Orthopaedics Network, Hamburg D-66421, Germany.
Mol Med Rep. 2021 Mar;23(3). doi: 10.3892/mmr.2021.11824. Epub 2021 Jan 5.
Stem cell‑based therapy is a promising alternative to conventional approaches to treating intervertebral disc degeneration (IDD). However, comprehensive understanding of stem cell‑based therapy at the gene level is still lacking. In the present study, we identified the expression profiles of messenger RNAs (mRNAs) and long non‑coding RNAs (lncRNAs) expressed within a co‑culture system of adipose‑derived mesenchymal stem cells (ASCs) and degenerative nucleus pulposus cells (NPCs) and explored the signaling pathways involved and their regulatory networks. Microarray analysis was used to compare ASCs co‑cultured with degenerative NPCs to ASCs cultured alone, and the underlying regulatory pattern, including the signaling pathways and competing endogenous RNA (ceRNA) network, was analyzed with robust bioinformatics methods. The results showed that 360 lncRNAs and 1757 mRNAs were differentially expressed by ASCs, and the microarray results were confirmed by quantitative PCR. Moreover, 589 Gene Ontology terms were upregulated, whereas 661 terms were downregulated. A total of 299 signaling pathways were significantly altered. A Path‑net and a Signal‑net were built to show interactions among differentially expressed genes. An mRNA‑lncRNA co‑expression network was constructed to reveal the interplay among differentially expressed mRNAs and lncRNAs, whereas a ceRNA network was built to investigate their connections with microRNAs involved in IDD. To the best of our knowledge, this original and comprehensive exploration reveals differentially expressed lncRNAs and mRNAs of ASCs stimulated by degenerative NPCs, underscoring the regulation pattern within the co‑culture system at the gene level. These data may further understanding of NPC‑directed differentiation of ASCs and facilitate the application of ASCs in future treatments for IDD.
基于干细胞的治疗是一种有前途的替代传统方法来治疗椎间盘退行性变(IDD)。然而,在基因水平上对基于干细胞的治疗的全面理解仍然缺乏。在本研究中,我们鉴定了脂肪间充质干细胞(ASCs)和退行性核髓细胞(NPCs)共培养体系中表达的信使 RNA(mRNA)和长非编码 RNA(lncRNA)的表达谱,并探讨了涉及的信号通路及其调控网络。微阵列分析用于比较与退行性 NPC 共培养的 ASC 与单独培养的 ASC,并使用强大的生物信息学方法分析潜在的调控模式,包括信号通路和竞争内源性 RNA(ceRNA)网络。结果显示,360 个 lncRNA 和 1757 个 mRNA 在 ASC 中差异表达,微阵列结果通过定量 PCR 得到验证。此外,589 个基因本体论术语上调,而 661 个术语下调。总共 299 个信号通路发生显著改变。构建了 Path-net 和 Signal-net 以显示差异表达基因之间的相互作用。构建了 mRNA-lncRNA 共表达网络,以揭示差异表达的 mRNA 和 lncRNA 之间的相互作用,而构建了 ceRNA 网络以研究它们与参与 IDD 的 microRNA 的连接。据我们所知,这项原创性和全面的探索揭示了退行性 NPC 刺激的 ASC 中差异表达的 lncRNA 和 mRNA,突出了共培养体系中基因水平的调控模式。这些数据可能进一步了解 NPC 指导的 ASC 分化,并促进 ASC 在未来 IDD 治疗中的应用。