牛卵丘细胞中与 DNA 双链断裂相关的非编码 RNA 竞争性内源性 RNA 网络。

DNA Double-Strand Break-Related Competitive Endogenous RNA Network of Noncoding RNA in Bovine Cumulus Cells.

机构信息

Department of Laboratory Animals, College of Animal Sciences, Jilin University, Changchun 130062, China.

Experimental Testing Center, Jilin Agricultural Science and Technology University, Jilin 132101, China.

出版信息

Genes (Basel). 2023 Jan 22;14(2):290. doi: 10.3390/genes14020290.

DOI:10.3390/genes14020290

PMID:36833217

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9956238/

Abstract

(1) Background: DNA double strand breaks (DSBs) are the most serious form of DNA damage that affects oocyte maturation and the physiological state of follicles and ovaries. Non-coding RNAs (ncRNAs) play a crucial role in DNA damage and repair. This study aims to analyze and establish the network of ncRNAs when DSB occurs and provide new ideas for next research on the mechanism of cumulus DSB. (2) Methods: Bovine cumulus cells (CCs) were treated with bleomycin (BLM) to construct a DSB model. We detected the changes of the cell cycle, cell viability, and apoptosis to determine the effect of DSBs on cell biology, and further evaluated the relationship between the transcriptome and competitive endogenous RNA (ceRNA) network and DSBs. (3) Results: BLM increased γH2AX positivity in CCs, disrupted the G1/S phase, and decreased cell viability. Totals of 848 mRNAs, 75 long noncoding RNAs (lncRNAs), 68 circular RNAs (circRNAs), and 71 microRNAs (miRNAs) in 78 groups of lncRNA-miRNA-mRNA regulatory networks, 275 groups of circRNA-miRNA-mRNA regulatory networks, and five groups of lncRNA/circRNA-miRNA-mRNA co-expression regulatory networks were related to DSBs. Most differentially expressed ncRNAs were annotated to cell cycle, p53, PI3K-AKT, and WNT signaling pathways. (4) Conclusions: The ceRNA network helps to understand the effects of DNA DSBs activation and remission on the biological function of CCs.

摘要

（1）背景：DNA 双链断裂（DSBs）是影响卵母细胞成熟和卵泡及卵巢生理状态的最严重的 DNA 损伤形式。非编码 RNA（ncRNA）在 DNA 损伤和修复中发挥着至关重要的作用。本研究旨在分析并建立 DSB 发生时 ncRNA 的网络，为下一步研究卵丘 DSB 的机制提供新的思路。

（2）方法：用博来霉素（BLM）处理牛卵丘细胞（CCs）构建 DSB 模型。我们检测细胞周期、细胞活力和细胞凋亡的变化，以确定 DSB 对细胞生物学的影响，进一步评估转录组与竞争内源性 RNA（ceRNA）网络和 DSBs 之间的关系。

（3）结果：BLM 增加了 CCs 中的 γH2AX 阳性，破坏了 G1/S 期，降低了细胞活力。在 78 组 lncRNA-miRNA-mRNA 调控网络、275 组 circRNA-miRNA-mRNA 调控网络和 5 组 lncRNA/circRNA-miRNA-mRNA 共表达调控网络中，共鉴定出 848 个 mRNAs、75 个长非编码 RNA（lncRNA）、68 个环状 RNA（circRNA）和 71 个 microRNA（miRNA）与 DSBs 相关。大多数差异表达的 ncRNA 被注释到细胞周期、p53、PI3K-AKT 和 WNT 信号通路。

（4）结论：ceRNA 网络有助于理解 DNA DSBs 激活和缓解对 CCs 生物学功能的影响。