Effects of Oxidative Stress on the MicroRNAs Profiles of EA.hy926 Cells and Bioinformatics Analysis Under Simulated Microgravity
Author(s): Liu Jia, Hu Taohong, Jia Mingwen, Zheng Hongwei, Yao Jing, Yuan Min, Wang Jingyu, Yuan Ming, State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center
Pages: 393-400
Year: 2019 Issue: 5
Journal: Space Medicine & Medical Engineering
Keyword: miRNA sequencing; vascular endothelial cells; simulated microgravity; oxidative stress;
Abstract: Objective To explore the differentially expressed microRNAs(miRNAs)profiles and their biological significance induced by oxidative stress in EA.hy926 cells under simulated microgravity induced by clinostat.Methods EA.hy926 cells were divided into the control group(Con),oxidative stress for control group(Con+H2 O2),simulated microgravity group(MG)and oxidative stress for simulated microgravity group(MG+H2 O2).The Illumina HiSeq 2500 platform sequencing technology was used to perform miRNA sequencing(miRNA-seq)and screen differentially expressed miRNAs.The selected differentially expressed miRNAs were verified by qRT-PCR technology and their predicted target genes were analyzed with Gene Ontology(GO)enrichment,KEGG pathway and STRING protein interaction analysis by online analysis tools.Results The miRNA profiles changed in EA.hy926 cells induced by oxidative stress under simulated microgravity.qRT-PCR showed the expression of hsa-miR-29 b-3 p and hsa-miR-200 c-3 p increased in MG+H2 O2 group compared with Con+H2 O2 group,which were consistent with the results of miRNA-seq.GO and KEGG pathway significant enrichment analysis showed that the target genes of hsa-miR-29 b-3 p were significantly enriched in the biological process of extracellular matrix organization and focal adhesion pathway,etc.Target genes of hsa-miR-200 c-3 p were significantly enriched in the biological process of regulation of cellular metabolic process and miRNA in cancer pathway,etc.Protein-protein interaction analysis results showed that proteins of COL family were in the key position of hsa-miR-29 b-3 p target genes,and RHOA protein was in the key position of hsa-miR-200 c-3 p target genes.Conclusion Oxidative stress under simulated microgravity induces significantly different expression of miRNAs in EA.hy926 cells.Hsa-miR-29 b-3 p and hsa-miR-200 c-3 p may participate in the dysfunction regulation of endothelial cells under microgravity by regulating target genes and pathways.
Pages: 393-400
Year: 2019 Issue: 5
Journal: Space Medicine & Medical Engineering
Keyword: miRNA sequencing; vascular endothelial cells; simulated microgravity; oxidative stress;
Abstract: Objective To explore the differentially expressed microRNAs(miRNAs)profiles and their biological significance induced by oxidative stress in EA.hy926 cells under simulated microgravity induced by clinostat.Methods EA.hy926 cells were divided into the control group(Con),oxidative stress for control group(Con+H2 O2),simulated microgravity group(MG)and oxidative stress for simulated microgravity group(MG+H2 O2).The Illumina HiSeq 2500 platform sequencing technology was used to perform miRNA sequencing(miRNA-seq)and screen differentially expressed miRNAs.The selected differentially expressed miRNAs were verified by qRT-PCR technology and their predicted target genes were analyzed with Gene Ontology(GO)enrichment,KEGG pathway and STRING protein interaction analysis by online analysis tools.Results The miRNA profiles changed in EA.hy926 cells induced by oxidative stress under simulated microgravity.qRT-PCR showed the expression of hsa-miR-29 b-3 p and hsa-miR-200 c-3 p increased in MG+H2 O2 group compared with Con+H2 O2 group,which were consistent with the results of miRNA-seq.GO and KEGG pathway significant enrichment analysis showed that the target genes of hsa-miR-29 b-3 p were significantly enriched in the biological process of extracellular matrix organization and focal adhesion pathway,etc.Target genes of hsa-miR-200 c-3 p were significantly enriched in the biological process of regulation of cellular metabolic process and miRNA in cancer pathway,etc.Protein-protein interaction analysis results showed that proteins of COL family were in the key position of hsa-miR-29 b-3 p target genes,and RHOA protein was in the key position of hsa-miR-200 c-3 p target genes.Conclusion Oxidative stress under simulated microgravity induces significantly different expression of miRNAs in EA.hy926 cells.Hsa-miR-29 b-3 p and hsa-miR-200 c-3 p may participate in the dysfunction regulation of endothelial cells under microgravity by regulating target genes and pathways.
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