miR-98 protects endothelial cells against hypoxia/reoxygenation induced-apoptosis by targeting caspase-3

• Published in: Biochemical and biophysical research communications Vol. 467; no. 3; pp. 595 - 601
• Main Authors: Li, He-wen, Meng, Yan, Xie, Qun, Yi, Wen-jing, Lai, Xue-li, Bian, Qi, Wang, Jun, Wang, Jia-feng, Yu, Guang
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20.11.2015
• Subjects: 3' Untranslated Regions; Animals; Apoptosis; Caspase 3 - genetics; Caspase 3 - metabolism; Caspase-3; Cell Hypoxia; Endothelium, Vascular - enzymology; Endothelium, Vascular - metabolism; Endothelium, Vascular - pathology; Human Umbilical Vein Endothelial Cells; Humans; Ischemia reperfusion injury; Male; Mice; Mice, Inbred C57BL; MicroRNAs - physiology; miR-98; Oxygen - administration & dosage; miR-98; Ischemia reperfusion injury; Caspase-3; Apoptosis
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2015.09.058

Endothelial dysfunction is one of the main pathophysiological processes involved in renal ischemia reperfusion injury. Our previous microarray study demonstrated that miR-98 was upregulated in the kidney with ischemia reperfusion injury (IRI). The present study was performed to investigate whether miR-98 was involved in the regulation of endothelial apoptosis under hypoxia and re-oxygenation (H/R) conditions. The dynamic changes of miR-98 in mouse IRI kidney and H/R HUVECs was measured. HUVECs were treated with HIF-1α siRNA to investigate the role of HIF-1α on miR-98 expression. The potential target genes of miR-98 were predicted by bioinformatics analyses. HUVECs were transfected with miR-98 mimics or inhibitor to confirm the role of miR-98 on the expression of target genes and hypoxia-induced apoptosis. The target gene was finally confirmed by dual-luciferase reporter assay. Both of IRI and H/R induced significantly up-regulation of miR-98 in the ischemic kidney and hypoxic HUVECs. HIF-1α siRNA remarkably down-regulated the expression of miR-98 in both normal and hypoxic HUVECs. The putative target genes of miR-98 included IL-6, IL-10 and caspase-3. MiR-98 mimics significantly inhibit caspase-3 expression in HUVECs, while anti-miR-98 significantly up-regulated it. But no change of IL-6 and IL-10 levels was observed after miRNA transfection. miR-98 protected HUVECs against apoptosis induced by hypoxia, while anti-miR-98 had the reverse effect. Furthermore, the dual-luciferase reporter assay confirmed that miR-98 decreased the luciferase activity by targeting the 3′ untranslated region of caspase-3. In conclusion, Renal IRI induces up-regulation of miR-98 dependent on HIF-1α, which protects endothelial cells against apoptosis by targeting caspase-3. •miR-98 is up-regulated in HUVECs with hypoxia/reoxygenation dependent on HIF-1α.•miR-98 upregulation inhibits hypoxia-induced apoptosis in endothelial cells.•miR-98 targets at caspase-3 3′UTR with high conservation among different species.