环状 RNA 在牙髓间充质干细胞修复受损子宫内膜的过程中大量表达并上调。
Circular RNAs are abundantly expressed and upregulated during repair of the damaged endometrium by Wharton's jelly-derived mesenchymal stem cells.
机构信息
Department of Obstetrics and Gynecology, Affiliated Hospital of Nantong University, Nantong, People's Republic of China.
Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Soochow University, Soochow, People's Republic of China.
出版信息
Stem Cell Res Ther. 2018 Nov 15;9(1):314. doi: 10.1186/s13287-018-1046-3.
BACKGROUND
Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) exhibit strong and powerful potential in repairing different diseases. The expression profile of circular RNA (circRNA) provides valuable insight for regulation of the repair process and exploration of reparative effect mechanisms.
METHODS
Human endometrial stromal cells (ESCs) were cultured with mifepristone to obtain damaged ESCs, which were then cocultured with or without WJ-MSCs (cocultured group versus non-cocultured group) to observe the reparative effect upon damaged ESCs by WJ-MSCs. CircRNA microarray was performed between the two groups. Based on the transcriptomics data, the differential gene expression profiles of the two groups were analyzed by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and network analysis methods. Screening of a circRNA database was performed, and the results were confirmed by quantitative polymerase chain reaction (qPCR).
RESULTS
WJ-MSCs exerted a reparative effect upon damaged ESCs in the cocultured group such as improved cell morphology, higher proliferative ability, and lower apoptosis rate. CircRNA array showed that 7757 circRNAs were differentially expressed in ESCs from the cocultured group. Mitotic cell cycle, cell cycle process, and nuclear division ranked top in the GO upregulated list of the two groups, while DNA replication and cell cycle ranked top in the KEGG pathway analysis upregulated list of the two groups. The nine most aberrantly expressed circRNAs were selected for further verification in the same cohort of samples by microarray analysis. Seven of the nine most aberrantly circRNAs were confirmed to be significantly upregulated in the cocultured group. And four of the seven circRNAs (hsa_circ_0015825, hsa-circRNA4049-38, hsa-circRNA5028-15, and hsa_circ_0111659) expression both in ESCs and WJ-MSCs tended to decrease with time by qPCR. The levels of the remaining three circRNAs (hsa-circRNA8881-21, hsa_circ_0020492 and hsa_circ_ 0026141) did not change significantly over time in either ESCs or WJ-MSCs. Moreover, we focused on hsa_circRNA_0111659 and predicted its miRNAs and targeted mRNA. The association of circRNA-miRNA-mRNA is likely to be involved in regulating the repair of endometrial damage.
CONCLUSIONS
Our results presented the abundant and upregulated circRNAs profile during repair of the damaged endometrium by WJ-MSCs and provided a novel perspective for circRNAs in the regulation of WJ-MSCs for endometrial repair.
背景
华通氏胶来源的间充质干细胞（WJ-MSCs）在修复多种疾病方面表现出强大的潜力。环状 RNA（circRNA）的表达谱为修复过程的调控和探索修复作用机制提供了有价值的见解。
方法
用米非司酮培养人子宫内膜基质细胞（ESCs）以获得受损的 ESCs，然后将其与 WJ-MSCs 共培养（共培养组与非共培养组），观察 WJ-MSCs 对受损 ESCs 的修复作用。对两组间进行环状 RNA 微阵列。基于转录组学数据，通过基因本体论（GO）、京都基因与基因组百科全书（KEGG）通路和网络分析方法分析两组间差异基因表达谱。对环状 RNA 数据库进行筛选，并通过定量聚合酶链反应（qPCR）进行验证。
结果
WJ-MSCs 在共培养组中对受损的 ESCs 发挥了修复作用，如改善细胞形态、提高增殖能力和降低凋亡率。环状 RNA 阵列显示，共培养组的 ESCs 中有 7757 个环状 RNA 差异表达。GO 上调列表中，有丝分裂细胞周期、细胞周期过程和核分裂排名最高，KEGG 通路分析上调列表中，DNA 复制和细胞周期排名最高。选择最显著差异表达的 9 个环状 RNA 进行同一队列样本的微阵列分析进一步验证。9 个最显著差异表达的环状 RNA 中有 7 个在共培养组中被证实显著上调。其中 4 个环状 RNA（hsa_circ_0015825、hsa-circRNA4049-38、hsa-circRNA5028-15 和 hsa_circ_0111659）在 ESCs 和 WJ-MSCs 中的表达均随时间呈下降趋势，通过 qPCR 验证。其余 3 个环状 RNA（hsa-circRNA8881-21、hsa_circ_0020492 和 hsa_circ_0026141）在 ESCs 或 WJ-MSCs 中随时间变化不明显。此外，我们专注于 hsa_circRNA_0111659，并预测其 miRNA 和靶向 mRNA。环状 RNA-miRNA-mRNA 的关联可能参与调控子宫内膜损伤的修复。
结论
我们的研究结果呈现了 WJ-MSCs 修复受损子宫内膜过程中丰富且上调的环状 RNA 图谱，为环状 RNA 在调控 WJ-MSCs 修复子宫内膜提供了新的视角。
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