血管内皮生长因子A调控人脐静脉内皮细胞miRNA的全基因表达谱分析

doi:10.3969/j.issn.1674-8115.2021.09.008

摘要/Abstract

摘要：

目的·分析人脐静脉内皮细胞（human umbilical vein endothelial cell，HUVEC）中微小RNA（microRNA，miRNA）在血管内皮生长因子A（vascular endothelial growth factor A，VEGFA）刺激下的变化，探索miRNA在调节VEGFA下游基因表达变化中的作用，并寻找新的血管新生调控miRNA。方法·通过NanoString nCounter分别检测了HUVEC在VEGFA刺激后0、1、4、12 h时miRNA的表达量，将具有相同变化趋势的差异表达miRNA聚类并预测其靶基因。通过基因本体数据库富集分析和京都基因与基因组百科全书通路分析，预测差异表达miRNA靶基因的相关功能，并建立互作网络。结果·VEGFA刺激HUVEC后，有39个miRNA的表达量发生了显著差异性变化。生物信息学分析结果显示差异表达miRNA所对应的靶mRNA有129个，动态表达的miRNA与其靶基因间的调控参与了血管新生中多个生物过程和信号通路。结论·VEGFA刺激诱导HUVEC差异性表达miRNA，miRNA在调节血管新生中具有重要作用；其中，miR-107和miR-21可能是血管新生的调控因子。

关键词: 微小RNA, 血管内皮生长因子A, 人脐静脉内皮细胞, 生物信息学分析

Abstract:

Objective·To analyze the changes of microRNA (miRNA) in human umbilical vein endothelial cells (HUVECs) under the stimulation of vascular endothelial growth factor A (VEGFA), in order to further explore the roles of miRNA in regulating the expression of downstream genes of VEGFA, and to find new regulatory miRNAs for angiogenesis.

Methods·The miRNA expression profile was analyzed by using NanoString nCounter in HUVECs 0, 1, 4, and 12 h after VEGFA stimulation. The differentially expressed miRNAs with the same trend were clustered and their target genes were predicted. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze and predict the functions of these target genes, and the interaction networks were established.

Results·The expression of 39 miRNAs changed significantly after VEGFA stimulation in HUVECs. Bioinformatic analysis showed that there were 129 target mRNAs corresponding to the differentially expressed miRNAs which were involved in multiple biological processes and signal pathways related to angiogenesis.

Conclusion·miRNAs are differentially expressed in HUVECs after stimulation by VEGFA, and miRNAs play important roles in regulating angiogenesis. MiR-107 and miR-21 may severe as candidate functional regulators of angiogenesis.

Key words: microRNA (miRNA), vascular endothelial growth factor A (VEGFA), human umbilical vein endothelial cell (HUVEC), bioinformatic analysis

中图分类号:

刘俐, 耿子龙, 陈嘉焕, 张沙沙, 张冰. 血管内皮生长因子A调控人脐静脉内皮细胞miRNA的全基因表达谱分析[J]. 上海交通大学学报(医学版), 2021, 41(9): 1183-1189.

Li LIU, Zi-long GENG, Jia-huan CHEN, Sha-sha ZHANG, Bing ZHANG. Whole gene expression profile analysis of miRNAs in human umbilical vein endothelial cells regulated by vascular endothelial growth factor A[J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(9): 1183-1189.

图/表 5

图1 miDEGs的靶基因

Fig 1 Target genes of miDEGs

图2 差异性表达miRNAs（miDEGs）的热图Note： Adjusted P value<0.01. Pearson′s r of counts 0 h, counts 1 h, counts 4 h and counts 12 h are 0.999 905 4, 0.999 899 1, 0.999 587 0, 0.999 643 5, respectively.

Fig 2 Heat map of differentially expressed miRNAs (miDEGs)

图3 miDEGs靶基因GO功能分析

Fig 3 GO function analysis of target genes of miDEGs

图4 miDEGs靶基因的KEGG通路分析

Fig 4 KEGG pathway analysis of target genes of miDEGs

图5 差异性表达miRNAs和它们靶基因的互作网络Note： HK2—hexokinase 2; PPARA—peroxisome proliferator activated receptor alpha; TNFAIP3—TNF alpha induced protein 3; BCL6—BCL6 transcription repressor; PRDM1—PR/SET domain 1; MXD1—MAX Dimerization Protein 1; ETS1—ETS proto-oncogene 1; PIK3R1—phosphoinositide-3-kinase regulatory subunit 1; KAT2B—lysine acetyltransferase 2B; BBC3—BCL2 binding component 3; MYCN—MYCN proto-oncogene, bHLH transcription factor; SIRT1—sirtuin 1; ICAM1—intercellular adhesion molecule 1; NFIA—nuclear factor I A; MCL1—MCL1 apoptosis regulator; CDK6—cyclin dependent kinase 6; DNMT3B—DNA methyltransferase 3 beta; IL6—interleukin 6; DDIT4—DNA damage inducible transcript 4; TMED7—transmembrane p24 trafficking protein 7; PAK1—p21 (RAC1) activated kinase 1.

Fig 5 Network of miDEGs and their target genes

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