The Second Clinical Medical College, Guangdong Medical University, Dongguan, 523808, Guangdong, China.
College of Life Science and Technology, Dalian University, Dalian, 116622, Liaoning, China.
J Transl Med. 2019 Oct 1;17(1):327. doi: 10.1186/s12967-019-2078-x.
Oxidative stress has a determinantal effect on human dental pulp stromal cells (hDPSCs), including affecting their longevity and functionality. Circular RNAs (circRNAs) play an essential role in stromal cell behavior; however, the exact mechanism in which circRNAs functions within hDPSCs were undergoing oxidative stress remains unclear. The purpose of this study is to assess the global changes and characteristics of circRNAs in hDPSCs undergoing oxidative stress.
Using an oxidative stress model of hDPSCs, we applied microarray analysis to examine the circRNAs profiles. We confirmed the changes in circRNAs by quantitative Real-Time PCR (qRT-PCR). Furthermore, bioinformatics tools, including a miRcode map, TargetScan, gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, were reconstructed for further assessment. SIRT1 gene and protein expression were tested by qRT-PCR and In Cell-Western analysis.
We revealed 330 upregulated, and 533 downregulated circRNAs undergoing oxidative stress in hDPSCs and confirmed three circRNAs distinct expressions (hsa_circ_0000257, hsa_circ_0087354, and hsa_circ_0001946) in hDPSCs undergoing oxidative stress by qRT-PCR. GO, and KEGG pathway enrichment revealed the differentially expressed circRNAs might participate in p53 and cell cycle signaling networks associated with oxidative stress. SIRT1 gene and protein expression was reduced in the oxidatively stressed cells (OSC) group compared to untreated cells (UC).
The findings of this study has provided new insights into circRNAs and a basis for further studies assessing the potential functions of hsa_circ_0000257, hsa_circ_0087354, and hsa_circ_0001946 in oxidatively stressed hDPSCs.
氧化应激对人牙髓基质细胞（hDPSCs）有决定性影响，包括影响其寿命和功能。环状 RNA（circRNA）在基质细胞行为中起着至关重要的作用；然而，circRNA 在 hDPSCs 中发挥作用的确切机制仍不清楚。本研究旨在评估氧化应激下人牙髓基质细胞中 circRNA 的整体变化和特征。
我们应用氧化应激模型的人牙髓基质细胞，通过微阵列分析来检测 circRNA 谱。我们通过定量实时 PCR（qRT-PCR）来验证 circRNA 的变化。此外，我们还使用生物信息学工具，包括 miRcode 图谱、TargetScan、基因本体论（GO）分析、京都基因与基因组百科全书（KEGG）通路分析，进行进一步评估。通过 qRT-PCR 和 In Cell-Western 分析测试 SIRT1 基因和蛋白表达。
我们发现 330 个上调和 533 个下调的 circRNA 在 hDPSCs 中经历氧化应激，并通过 qRT-PCR 证实了三个在 hDPSCs 中经历氧化应激的 circRNA 表达差异（hsa_circ_0000257、hsa_circ_0087354 和 hsa_circ_0001946）。GO 和 KEGG 通路富集显示，差异表达的 circRNA 可能参与与氧化应激相关的 p53 和细胞周期信号网络。与未处理细胞（UC）相比，氧化应激细胞（OSC）组中 SIRT1 基因和蛋白表达降低。
本研究的结果为 circRNA 提供了新的见解，并为进一步研究评估 hsa_circ_0000257、hsa_circ_0087354 和 hsa_circ_0001946 在氧化应激下人牙髓基质细胞中的潜在功能提供了基础。
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