Abstract
Endothelial cells are important components of peripheral nerve stumps that contribute to Schwann cell migration and peripheral nerve regeneration. Let-7d modulates the phenotype of Schwann cells and affected peripheral nerve regeneration. However, the regulatory roles of let-7d on endothelial cells remain undetermined. In this study, by transfecting cultured human umbilical vein endothelial cells (HUVECs) with let-7d mimic or let-7d inhibitor, we investigated the biological effects of let-7d on endothelial cells. EdU proliferation assay showed that upregulated let-7d decreased the proliferation rates of HUVECs while downregulated let-7d increased the proliferation rates of HUVECs. Transwell-based migration assay and wound-healing assay demonstrated that let-7d inhibited the migration ability of HUVECs. Matrigel assay suggested that let-7d decreased the numbers of formed meshes and suppressed the tubulogenesis of HUVECs. RNA sequencing, bioinformatic analysis, gene expression validation, and luciferase assay suggested that let-7d directly targeted interferon-induced protein 44 like (IFI44L) gene and negatively regulated the expression of IFI44L. Taken together, our study illuminated the inhibitory roles of let-7d on the proliferation, migration, and tubulogenesis of endothelial cells, identified the target gene of let-7d, and deepened the understanding of the biological effects of let-7d on key elements of peripheral nerve regeneration.
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Acknowledgements
This study was supported by National Natural Science Foundation of China (Grant No. 31700926) and Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD). The authors are grateful for Dr. Mi Shen for providing human umbilical vein endothelial cells (HUVECs).
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SY: conceived and designed the experiments. XJ, HH, SY and YS: performed the experiments. XJ, HH, SB and SY: analyzed the data. BS and SY: contributed reagents/materials/analysis tools. XJ and SY: wrote the manuscript.
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11010_2019_3611_MOESM1_ESM.xlsx
Supplementary material 1—List of potential targets of let-7d predicted by Miranda, RNAhybrid, and TargetScan. (XLSX 6819 kb)
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Ji, X., Hua, H., Shen, Y. et al. Let-7d modulates the proliferation, migration, tubulogenesis of endothelial cells. Mol Cell Biochem 462, 75–83 (2019). https://doi.org/10.1007/s11010-019-03611-x
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DOI: https://doi.org/10.1007/s11010-019-03611-x