Identification of the Transcriptional Networks and the Involvement in Angiotensin II-Induced Injury after CRISPR/Cas9-Mediated Knockdown of Cyr61 in HEK293T Cells
Identification of the Transcriptional Networks and the Involvement in Angiotensin II-Induced Injury after CRISPR/Cas9-Mediated Knockdown of Cyr61 in HEK293T Cells
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HEK293T 细胞中 CRISPR/Cas9 介导的 Cyr61 敲低后转录网络及其参与血管紧张素 II 诱导损伤的鉴定
DOI：
10.1155/2019/8697257
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发表时间：
2019-04
影响因子：
4.6
通讯作者：
You Yanwu
中科院分区：
文献类型：
--
作者：
Wang Junjie;Fu Dongdong;Senouthai Soulixay;Jiang Yan;Hu Rentong;You Yanwu
背景Cyr 61的转录网络及其在细胞损伤中的功能尚不清楚。本研究描述了在HEK 293 T细胞中由CRISPR/Cas9介导的Cyr 61敲低后lncRNA和mRNA谱以及参与血管紧张素II诱导的损伤。方法培养HEK 293 T细胞，通过转染CRISPR/Cas9 KO质粒实现Cyr 61敲低。lncRNA和mRNA微阵列用于鉴定差异表达基因（DEG）。进行基因本体论（GO）和京都基因和基因组百科全书（KEGG）途径分析以确定生物功能和信号传导途径。用RT-PCR验证芯片结果。将细胞分为四组：对照组、Cyr 61敲低组、无Cyr 61敲低的血管紧张素II（Ang II）组和Cyr 61敲低的Ang II组。进行CCK 8、蛋白质印迹和流式细胞术分析以剖析细胞功能。结果共归一化23184个lncRNA和28264个mRNA。Cyr 61基因敲除后，26种lncRNA和212种mRNA表达上调，74种lncRNA和233种mRNA表达下调。细胞成分，分子功能，生物学过程和调控途径与差异表达的mRNA的分析揭示了Cyr 61基因的下游机制。差异表达基因影响小细胞肺癌、轴突导向、Fc γ R介导的吞噬作用、MAPK信号通路、粘着斑、胰岛素抵抗和代谢通路。此外，Cyr 61的表达增加与Ang II诱导的细胞周期阻滞和凋亡的诱导以及细胞增殖的抑制一致。敲低Cyr 61基因可促进HEK 293 T细胞的细胞周期进程，减少细胞凋亡，促进细胞增殖。结论Cyr 61基因参与了Ang II诱导的HEK 293 T细胞损伤。对这些差异表达的lncRNA和mRNAs的功能机制以及代谢途径的研究将为Cyr 61相关疾病的治疗提供新的靶点。
Background The transcriptional networks of Cyr61 and its function in cell injury are poorly understood. The present study depicted the lncRNA and mRNA profiles and the involvement in angiotensin II-induced injury after Cyr61 knockdown mediated by CRISPR/Cas9 in HEK293T cells. Methods HEK293T cells were cultured, and Cyr61 knockdown was achieved by transfection of the CRISPR/Cas9 KO plasmid. lncRNA and mRNA microarrays were used to identify differentially expressed genes (DEGs). Gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to determine biofunctions and signaling pathways. RT-PCR was used to validate the microarray results. Cells were divided into four groups: control, Cyr61 knockdown, angiotensin II (Ang II) without Cyr61 knockdown, and Ang II with Cyr61 knockdown. CCK8, western blotting, and flow cytometry analysis were carried out to dissect cellular function. Results A total of 23184 lncRNAs and 28264 mRNAs were normalized. 26 lncRNAs and 212 mRNAs were upregulated, and 74 lncRNAs and 233 mRNAs were downregulated after Cyr61 knockdown. Analysis of cellular components, molecular functions, biological processes, and regulatory pathways associated with the differentially expressed mRNAs revealed downstream mechanisms of the Cyr61 gene. The differentially expressed genes were affected for small cell lung cancer, axon guidance, Fc gamma R-mediated phagocytosis, MAPK signaling pathway, focal adhesion, insulin resistance, and metabolic pathways. In addition, Cyr61 expression was increased in accordance with induction of cell cycle arrest and apoptosis and inhibition of cell proliferation induced by Ang II. Knockdown of Cyr61 in HEK293T cells promoted cell cycle procession, decreased apoptosis, and promoted cell proliferation. Conclusions The Cyr61 gene is involved in Ang II-induced injury in HEK293T cells. Functional mechanisms of the differentially expressed lncRNAs and mRNAs as well as identification of metabolic pathways will provide new therapeutic targets for Cyr61-realated diseases.
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影响因子：
17.1
作者：
Pinter M;Jain RK
通讯作者：
Jain RK
DOI：
10.1002/(sici)1097-0045(19980701)36:2
发表时间：
1998-07
期刊：
The Prostate
影响因子：
--
作者：
C. Pilarsky;U. Schmidt;Claudia Eißrich;Jörg Stade;S. Froschermaier;M. Haase;G. Faller;Thomas W. Kirchner;M. Wirth
通讯作者：
C. Pilarsky;U. Schmidt;Claudia Eißrich;Jörg Stade;S. Froschermaier;M. Haase;G. Faller;Thomas W. Kirchner;M. Wirth
影响因子：
5.6
作者：
Li XC;Zhuo JL
通讯作者：
Zhuo JL
CCN1/Cyr61在女性生殖系统胚胎发生和肿瘤发生中的生物学功能和作用
影响因子：
3.4
作者：
Yang R;Chen Y;Chen D
通讯作者：
Chen D
血管紧张素 II 下调 nephrin-Akt 信号传导并诱导足细胞损伤：c-Abl 的作用
影响因子：
3.3
作者：
Yang Q;Ma Y;Liu Y;Liang W;Chen X;Ren Z;Wang H;Singhal PC;Ding G
通讯作者：
Ding G