构建多柔比星诱导心肌损伤中铁死亡调控的 ceRNA 网络。

Construction of a ceRNA network of regulated ferroptosis in doxorubicin-induced myocardial injury.

机构信息

Department of Physiology, Bengbu Medical College, Bengbu, Anhui, China.

Key Laboratory of Basic and Clinical Cardiovascular Diseases, Bengbu Medical College, Bengbu, Anhui, China.

出版信息

PeerJ. 2023 Jan 25;11:e14767. doi: 10.7717/peerj.14767. eCollection 2023.

DOI:10.7717/peerj.14767

PMID:36718444

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9884038/

Abstract

BACKGROUND

Ferroptosis and long-noncoding RNAs (lncRNAs) play crucial roles in doxorubicin (DOX)-induced myocardial injury (DIMI). Nevertheless, there is no research to construct competing endogenous RNAs (ceRNAs) network between lncRNAs and ferroptosis-related key gene. So our research was designed to screen ferroptosis-related genes from differentially expressed mRNAs in DIMI and construct lncRNAs regulated ferroptosis-related key gene ceRNAs network.

METHODS

The male mice were injected with DOX intraperitoneally to induce myocardial injury, myocardial injury was evaluated by hematoxylin and eosin (HE) staining, and ferroptosis-related protein-glutathione peroxidase 4 (GPx4) protein expression was detected. The differentially expressed lncRNAs and mRNAs were detected by microarray, and the ferroptosis-related genes were screened to construct a protein-protein associations (PPA) network, the highest maximal clique centrality (MCC) score gene were identified by Cytoscape software, miRNAs bound to key genes and lncRNAs bound to miRNAs were predicted; then, the obtained lncRNAs were intersected with differentially expressed lncRNAs detected by microarray. Finally, the lncRNA/miRNA/mRNA ceRNA network of the highest MCC score gene regulating ferroptosis in DIMI was constructed. The expressions of the key components in ceRNA network were detected by qRT-PCR.

RESULTS

Compared with the control group, in the DOX group, myocardial enzymes and HE staining showed that myocardium structure was changed, and GPx4 protein expression was decreased. The differentially expressed 10,265 lncRNAs and 6,610 mRNAs in the DOX group were detected microarray. Among them, 114 ferroptosis-related genes were obtained to construct PPA networks, and Becn1 was identified as the key gene. Finally, the ceRNA network including Becn1, three miRNAs and four lncRNAs was constructed by predicting data of the Starbase database. The relative expressions of these components in ceRNA net were up-regulated and consistent with microarray results.

CONCLUSIONS

Based on the microarray detection results and bioinformatics analysis, we screened ferroptosis-related gene Becn1 and constructed the lncRNA/miRNA/mRNA ceRNA network of regulated ferroptosis in DIMI.

摘要

背景

铁死亡和长链非编码 RNA（lncRNA）在多柔比星（DOX）诱导的心肌损伤（DIMI）中发挥着重要作用。然而，目前还没有研究构建 lncRNA 与铁死亡相关关键基因之间的竞争性内源 RNA（ceRNA）网络。因此，我们的研究旨在从 DIMI 中差异表达的 mRNA 中筛选铁死亡相关基因，并构建 lncRNA 调控铁死亡相关关键基因 ceRNA 网络。

方法

雄性小鼠腹腔注射 DOX 诱导心肌损伤，通过苏木精和伊红（HE）染色评估心肌损伤，检测铁死亡相关蛋白-谷胱甘肽过氧化物酶 4（GPx4）蛋白表达。通过微阵列检测差异表达的 lncRNA 和 mRNA，筛选铁死亡相关基因构建蛋白质-蛋白质相互作用（PPA）网络，使用 Cytoscape 软件识别最高最大团中心度（MCC）评分基因，预测与关键基因结合的 miRNA 和与 miRNA 结合的 lncRNA；然后，将获得的 lncRNA 与微阵列检测到的差异表达 lncRNA 进行交集。最后，构建 DIMI 中调节铁死亡的最高 MCC 评分基因的 lncRNA/miRNA/mRNA ceRNA 网络。通过 qRT-PCR 检测 ceRNA 网络中关键成分的表达。

结果

与对照组相比，DOX 组心肌酶和 HE 染色显示心肌结构发生改变，GPx4 蛋白表达降低。通过微阵列检测到 DOX 组差异表达的 10265 个 lncRNA 和 6610 个 mRNA。其中，获得 114 个铁死亡相关基因构建 PPA 网络，鉴定出 Becn1 为关键基因。最后，通过预测 Starbase 数据库的数据构建了包括 Becn1、三个 miRNA 和四个 lncRNA 的 ceRNA 网络。ceRNA 网络中这些成分的相对表达均上调，与微阵列结果一致。