Transcriptomic Analysis of Pulmonary Microvascular Endothelial Cells with IQGAP1 Knockdown.
Transcriptomic Analysis of Pulmonary Microvascular Endothelial Cells with IQGAP1 Knockdown.
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DOI：
10.1089/dna.2020.5451
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发表时间：
2020-05
影响因子：
3.1
通讯作者：
Shihong Su;Aihui Xu;Yang Chen;Wanzhen Li;Xiaojun Zha;Yani Wang;Gengyun Sun
中科院分区：
文献类型：
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作者：
Shihong Su;Aihui Xu;Yang Chen;Wanzhen Li;Xiaojun Zha;Yani Wang;Gengyun Sun
在急性呼吸窘迫综合征和急性肺损伤（ARDS/ALI）中，肺微血管内皮屏障在保护肺组织免受炎症损伤中起关键作用。iq - gtpase -激活蛋白1 （IQGAP1）的异常表达是内皮屏障损伤的重要病因。然而，由IQGAP1直接调控的显著差异表达基因（deg）和信号通路过于复杂，无法完全理解。本研究基于转录组RNA测序（RNA- seq），在大鼠肺微血管内皮细胞中共鉴定出1216个受IQGAP1敲低调控的deg。其中，665个基因表达上调，551个基因表达下调。基因本体论分析表明，上调的DEGs主要富集于DNA复制、细胞周期和染色体形成，下调的DEGs主要参与细胞增殖、细胞粘附、细胞迁移等多种细胞生物过程的调控。京都基因和基因组百科全书对DEGs的通路分析显示，上调的通路主要与DNA复制有关，而显著下调的通路主要与TNF信号通路和一些炎症和增殖相关的通路有关。此外，我们选择30℃进行qRT-PCR验证，结果与RNA-Seq相当一致。此外，我们还发现IQGAP1基因敲低对许多在ARDS/ALI中起重要作用的细胞因子和炎症因子有显著影响。综上所述，本研究在RNA-Seq的基础上，我们发现IQGAP1不仅在微血管内皮屏障中发挥重要作用，而且在炎症中也发挥重要作用，这可能为今后研究IQGAP1在ARDS/ALI等相关疾病中的作用提供新的思路。
Pulmonary microvascular endothelium barrier plays a critical role in protecting the pulmonary tissue from inflammatory injury in acute respiratory distress syndrome and acute lung injury (ARDS/ALI). The dysregulation of IQ-GTPase-activating protein 1 (IQGAP1) was an important etiology of endothelium barrier injury. However, significant differentially expressed genes (DEGs) and signaling pathways directly regulated by IQGAP1 are too complicated to fully understand. In this research, we identified a total of 1216 DEGs regulated by knockdown of IQGAP1 in rat pulmonary microvascular endothelial cells on the basis of transcriptomic RNA sequencing (RNA-Seq). Among them, 665 were upregulated DEGs and 551 were downregulated DEGs. Gene ontology analysis has revealed that upregulated DEGs were mainly enriched in DNA replication, cell cycle, and chromosome formation, while downregulated DEGs were mainly involved in the regulation of many cellular bioprocesses including cell proliferation, cell adhesion, and cell migration. Kyoto Encyclopedia of Genes and Genomes pathways analysis toward DEGs showed that upregulated pathways were mainly about DNA replication, while the significantly downregulated pathways were about TNF signaling pathway and some inflammatory- and proliferation-related pathways. Furthermore, we choose 30 DEGs for validation by qRT-PCR, the results were quite consistent with the RNA-Seq. In addition, we also found that knockdown of IQGAP1 caused a significant impact on many cytokines and inflammatory factors, which play a vital role in ARDS/ALI. In summary, in this study on the basis of RNA-Seq, we found IQGAP1 not only exerts a crucial role in microvascular endothelium barrier but also plays an important role in inflammation, which might provide a new insight for future study on IQGAP1 in the related diseases such as ARDS/ALI.