Analysis of Differentially Expressed Genes in Endothelial Cells Following Tumor Cell Adhesion, and the Role of PRKAA2 and miR-124-3p.
Analysis of Differentially Expressed Genes in Endothelial Cells Following Tumor Cell Adhesion, and the Role of PRKAA2 and miR-124-3p.
复制标题
肿瘤细胞粘附后内皮细胞差异表达基因分析以及 PRKAA2 和 miR-124-3p 的作用
DOI：
10.3389/fcell.2021.604038
复制
发表时间：
2021
影响因子：
5.5
通讯作者：
Li X
中科院分区：
文献类型：
--
作者：
Pan Y;Abdureyim M;Yao Q;Li X
肿瘤细胞与内皮细胞的粘附是肿瘤与内皮细胞相互作用的一种方式，也是肿瘤转移的关键步骤。内皮细胞的完整性是阻止肿瘤侵袭和转移的重要屏障。肿瘤细胞粘附引起的内皮细胞变化为肿瘤细胞的血管生成和转移提供了重要的信号机制。然而，当肿瘤与内皮细胞的相互作用尚不清楚时，这些变化发生在内皮细胞上。在本研究中，我们使用了Affyssin基因芯片Human Transcriptome Array 2.0。和定量实时PCR（qPCR）来阐明前列腺肿瘤细胞PC-3 M粘附的内皮细胞中详细的基因改变。通过芯片数据分析，共获得504条差异表达的mRNA和444条lncRNA。基因本体（GO）功能分析表明，差异表达基因（DEG）在生物学水平主要介导腺体发育和DNA复制;在细胞组分水平主要参与线粒体内膜;在分子功能水平主要富集ATP酶活性和催化活性。京都基因和基因组百科全书（KEGG）信号通路分析显示，DEG主要调节癌症、细胞周期、嘧啶代谢和mTOR信号通路中的通路。然后，我们使用Cytoscape v3.7.2构建了蛋白质-蛋白质相互作用功能网络和mRNA-lncRNA相互作用网络。来鉴定核心基因mRNA和lncRNA使用数据库（miRDB、RNA22和Targetscan）预测核心mRNA PRKAA 2靶向的miRNA。qPCR结果显示，在PC-3M粘附的内皮细胞中，PRKAA2的预测靶miRNA miR-124 - 3p显著下调。通过双荧光素酶报告基因测定，证实了miR-124 - 3p与PRKAA 2 3'UTR的结合。此外，通过使用miR-124 - 3p的敲低慢病毒载体来下调内皮细胞中miR-124 - 3p的表达水平，我们发现PRKAA2的表达水平相应地增加。综上所述，肿瘤细胞的粘附对内皮细胞中mRNA和lncRNA有显著影响，其中PRKAA 2是一个显著变化的分子，miR-124 - 3p可以调节其在内皮细胞中的表达和功能。
Tumor cell adhesion to the endothelium is one pattern of tumor–endothelium interaction and a key step during tumor metastasis. Endothelium integrity is an important barrier to prevent tumor invasion and metastasis. Changes in endothelial cells (ECs) due to tumor cell adhesion provide important signaling mechanisms for the angiogenesis and metastasis of tumor cells. However, the changes happened in endothelial cells when tumor–endothelium interactions are still unclear. In this study, we used Affymetrix Gene Chip Human Transcriptome Array 2.0. and quantitative real-time PCR (qPCR) to clarify the detailed gene alteration in endothelial cells adhered by prostate tumor cells PC-3M. A total of 504 differentially expressed mRNAs and 444 lncRNAs were obtained through chip data analysis. Gene Ontology (GO) function analysis showed that differentially expressed genes (DEGs) mainly mediated gland development and DNA replication at the biological level; at the cell component level, they were mainly involved in the mitochondrial inner membrane; and at the molecular function level, DEGs were mainly enriched in ATPase activity and catalytic activity. Kyoto Encyclopedia of Genes and Genomes (KEGG) signal pathway analysis showed that the DEGs mainly regulated pathways in cancer, cell cycle, pyrimidine metabolism, and the mTOR signaling pathway. Then, we constructed a protein–protein interaction functional network and mRNA–lncRNA interaction network using Cytoscape v3.7.2. to identify core genes, mRNAs, and lncRNAs. The miRNAs targeted by the core mRNA PRKAA2 were predicted using databases (miRDB, RNA22, and Targetscan). The qPCR results showed that miR-124-3p, the predicted target miRNA of PRKAA2, was significantly downregulated in endothelial cells adhered by PC-3M. With a dual luciferase reporter assay, the binding of miR-124-3p with PRKAA2 3’UTR was confirmed. Additionally, by using the knockdown lentiviral vectors of miR-124-3p to downregulate the miR-124-3p expression level in endothelial cells, we found that the expression level of PRKAA2 increased accordingly. Taken together, the adhesion of tumor cells had a significant effect on mRNAs and lncRNAs in the endothelial cells, in which PRKAA2 is a notable changed molecule and miR-124-3p could regulate its expression and function in endothelial cells.
登录
查看更多内容
HSP60 的下调通过 ROS/AMPK/mTOR 途径抑制胶质母细胞瘤细胞的增殖
影响因子：
4.6
作者：
Tang H;Li J;Liu X;Wang G;Luo M;Deng H
通讯作者：
Deng H
影响因子：
20.1
作者：
Kohlstedt, Karin;Trouvain, Caroline;Fleming, Ingrid
通讯作者：
Fleming, Ingrid
影响因子：
--
作者：
Liu, Zhong;Shao, Yongfu;Guo, Junming
通讯作者：
Guo, Junming
影响因子：
3.6
作者：
Liu Q;Wu DH;Han L;Deng JW;Zhou L;He R;Lu CJ;Mi QS
通讯作者：
Mi QS
CERNA假设：隐藏RNA语言的Rosetta石头？
影响因子：
64.5
作者：
Salmena L;Poliseno L;Tay Y;Kats L;Pandolfi PP
通讯作者：
Pandolfi PP