The Angiogenic Effect of microRNA-21 Targeting TIMP3 through the Regulation of MMP2 and MMP9

• Published in: PloS one Vol. 11; no. 2; p. e0149537
• Main Authors: Hu, Jianzhong, Ni, Shuangfei, Cao, Yong, Zhang, Tao, Wu, Tianding, Yin, Xianzhen, Lang, Ye, Lu, Hongbin
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 12.02.2016; Public Library of Science (PLoS)
• Subjects: Analysis; Angiogenesis; Animals; Apoptosis; Biology and Life Sciences; Brain research; Care and treatment; Cell culture; Cell Death; Cell Movement; Cell Survival; Computed tomography; Deprivation; Endothelial cells; Gelatinase A; Gelatinase B; Gene expression; Gene Expression Regulation; Glucose; Human Umbilical Vein Endothelial Cells; Injuries; Ischemia; Lung cancer; Matrix metalloproteinase; Matrix Metalloproteinase 2 - genetics; Matrix Metalloproteinase 2 - metabolism; Matrix Metalloproteinase 9 - genetics; Matrix Metalloproteinase 9 - metabolism; Medical imaging; Medical research; Medicine; Medicine and Health Sciences; Metalloproteinase; Metastasis; MicroRNA; MicroRNAs; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; Neovascularization, Physiologic; Oxygen; Radiation; Radiation injuries; Rats; Research and Analysis Methods; Ribonucleic acid; RNA; RNA Interference; RNA, Small Interfering - genetics; RNA-mediated interference; Rodents; Secretion; siRNA; Spinal cord injuries; Sports medicine; Surgery; Survival; Synchrotron radiation; Tissue inhibitor of metalloproteinase 3; Tissue Inhibitor of Metalloproteinase-3 - genetics; Tissue Inhibitor of Metalloproteinase-3 - metabolism; Vascular endothelial growth factor; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0149537

microRNAs are a novel set of small, non-protein-coding nucleotide RNAs that negatively regulate the expression of target mRNAs. miRNA-21 is a microRNA that is highly enriched in endothelial cells. miRNA-21 has been shown to be a potential pro-angiogenic factor in some biological systems. Our previous study showed that the expression of miRNA-21 was up-regulated after spinal cord injury. However, the effect of miRNA-21 on angiogenesis in the spinal cord was unclear. In this study, to understand the role of miRNA-21 on injured endothelial cells exclusively, an oxygen and glucose deprivation model of endothelial cells was constructed, and the up-regulation of miRNA-21 was discovered in this model. An increased level of miRNA-21 by mimics promoted the survival, migration and tube formation of endothelial cells, which simultaneously inhibited tissue inhibitor of metalloproteinase-3 (TIMP3) expression and promoted matrix metalloproteinase-2 (MMP2) and matrix metalloproteinase-9 (MMP9) expression and secretion. A decreased level of miRNA-21 by antagomir exerted an opposite effect. As is well known, survival, migration and tube formation of endothelial cells are necessary prerequisites for angiogenesis after injury. TIMP3 was validated as a direct target of miRNA-21 by dual-luciferase reporter assay. Silencing with small interfering RNA against TIMP3 promoted tube formation and increased MMP2 and MMP9 expression at the protein level. In vivo, we found that decreased levels of miRNA-21 inhibited angiogenesis after spinal cord injury in rats using synchrotron radiation micro-computed tomography. In summary, these findings suggest that miRNA-21 has a protective effect on angiogenesis by reducing cell death and promoting cell survival, migration and tube formation via partially targeting the TIMP3 by potentially regulating MMP2 and MMP9. TIMP3 is a functional target gene. Identifying the role of miRNA-21 in the protection of angiogenesis might offer a novel therapeutic target for secondary spinal cord injury, in which angiogenesis is indispensable.