hsa-miR-204-5p对人脐静脉内皮细胞生物学行为的影响
R543%R363.2%Q752; 目的:探究hsa-miR-204-5p对人血管内皮细胞活力、迁移、周期及凋亡等生物学行为的影响.方法:使用人脐静脉内皮细胞株EA.hy926构建过表达hsa-miR-204-5p(hsa-miR-204-5p mimics)模型.通过细胞划痕、Transwell、CCK-8、细胞周期和凋亡率等指标评估hsa-miR-204-5p对内皮细胞功能状态的影响.接着,使用RNA测序和RT-qPCR寻找并验证hsa-miR-204-5p的下游靶基因,将RNA测序中log2FC≤-0.5、P＜0.05的基因和miRWalk数据库预测的hsa-miR-204-5p的下游靶...
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| Published in | 中国病理生理杂志 Vol. 40; no. 2; pp. 230 - 237 |
|---|---|
| Main Authors | , , |
| Format | Journal Article |
| Language | Chinese |
| Published |
广西医科大学基础医学院病理生理学教研室,广西 南宁 530021
2024
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| Subjects | |
| Online Access | Get full text |
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| Abstract | R543%R363.2%Q752; 目的:探究hsa-miR-204-5p对人血管内皮细胞活力、迁移、周期及凋亡等生物学行为的影响.方法:使用人脐静脉内皮细胞株EA.hy926构建过表达hsa-miR-204-5p(hsa-miR-204-5p mimics)模型.通过细胞划痕、Transwell、CCK-8、细胞周期和凋亡率等指标评估hsa-miR-204-5p对内皮细胞功能状态的影响.接着,使用RNA测序和RT-qPCR寻找并验证hsa-miR-204-5p的下游靶基因,将RNA测序中log2FC≤-0.5、P＜0.05的基因和miRWalk数据库预测的hsa-miR-204-5p的下游靶基因取交集,并进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析.结果:过表达hsa-miR-204-5p可以抑制EA.hy926细胞的活力和迁移能力,降低EA.hy926细胞的凋亡率,并使聚集在S期的细胞减少.富集分析结果发现hsa-miR-204-5p下游靶基因富集在MAPK信号通路,富集在此通路的基因为MAPT、PPP3R1、PRKACB、PTPRR、MAP2K4、CACNA2D2和RPS6KA6.RT-qPCR结果显示,过表达hsa-miR-204-5p后,MAPT和MAP2K4扩增结果较好且表达为下调.其中,MAPT下降趋势最明显.结论:hsa-miR-204-5p可能通过抑制MAPT/MAPK信号通路而影响内皮细胞的活力、迁移和凋亡等生物学行为. |
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| AbstractList | R543%R363.2%Q752; 目的:探究hsa-miR-204-5p对人血管内皮细胞活力、迁移、周期及凋亡等生物学行为的影响.方法:使用人脐静脉内皮细胞株EA.hy926构建过表达hsa-miR-204-5p(hsa-miR-204-5p mimics)模型.通过细胞划痕、Transwell、CCK-8、细胞周期和凋亡率等指标评估hsa-miR-204-5p对内皮细胞功能状态的影响.接着,使用RNA测序和RT-qPCR寻找并验证hsa-miR-204-5p的下游靶基因,将RNA测序中log2FC≤-0.5、P＜0.05的基因和miRWalk数据库预测的hsa-miR-204-5p的下游靶基因取交集,并进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析.结果:过表达hsa-miR-204-5p可以抑制EA.hy926细胞的活力和迁移能力,降低EA.hy926细胞的凋亡率,并使聚集在S期的细胞减少.富集分析结果发现hsa-miR-204-5p下游靶基因富集在MAPK信号通路,富集在此通路的基因为MAPT、PPP3R1、PRKACB、PTPRR、MAP2K4、CACNA2D2和RPS6KA6.RT-qPCR结果显示,过表达hsa-miR-204-5p后,MAPT和MAP2K4扩增结果较好且表达为下调.其中,MAPT下降趋势最明显.结论:hsa-miR-204-5p可能通过抑制MAPT/MAPK信号通路而影响内皮细胞的活力、迁移和凋亡等生物学行为. |
| Abstract_FL | AIM:This study aimed to investigate the effects of hsa-miR-204-5p on the viability,migration,cell cycle,and apoptosis of human vascular endothelial cells.METHODS:We established a model using the hsa-miR-204-5p mimic in the human umbilical vein endothelial cell line EA.hy926.We evaluated the effects of hsa-miR-204-5p on endothelial cell functionality through various analyses,including cell scratch,Transwell,CCK-8,cell cycle,and apopto-sis assays.Subsequently,we employed RNA sequencing and RT-qPCR to predict and verify the downstream target genes of hsa-miR-204-5p.Genes meeting the criteria of log2FC≤-0.5 and P＜0.05 in RNA sequencing and those predicted as downstream target genes of hsa-miR-204-5p by the miRWalk database were intersected.Furthermore,we conducted Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.RESULTS:Overexpres-sion of hsa-miR-204-5p inhibited the viability and migration of EA.hy926 cells,and reduced their apoptotic rate and the proportion of cells in S phase.Enrichment analyses showed that downstream target genes of hsa-miR-204-5p,including MAPT,PPP3R1,PRKACB,PTPRR,MAP2K4,CACNA2D2 and RPS6KA6,exhibited enrichment in MAPK signaling pathway.RT-qPCR results revealed that the mRNA expression levels of MAPT and MAP2K4,especially MAPT,were sig-nificantly down-regulated after overexpression of hsa-miR-204-5p.CONCLUSION:The findings suggest that hsa-miR-204-5p suppresses the biological behaviors of endothelial cells,such as viability,migration,and apoptosis,likely through the inhibition of MAPT/MAPK signaling pathway. |
| Author | 杨英 张义炜 潘尚领 |
| AuthorAffiliation | 广西医科大学基础医学院病理生理学教研室,广西 南宁 530021 |
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| Author_FL | ZHANG Yiwei YANG Ying PAN Shangling |
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| DocumentTitle_FL | Effects of hsa-miR-204-5p on biological behaviors of human umbilical vein endothelial cells |
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| Keywords | MAPK signaling pathway hsa-miR-204-5p MAPK信号通路 vascular endothelial cells 血管内皮细胞 RNA测序 RNA seqencing |
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| Title | hsa-miR-204-5p对人脐静脉内皮细胞生物学行为的影响 |
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