miR-4463 Regulates Hypoxia-Induced Autophagy and Apoptosis by Targeting ULK1 in Endothelial Cells
Background: Our previous study revealed aberrant miR-4463 expression in the vascular tissues of patients with arteriosclerosis obliterans of the lower extremities (ASO), but the role of miR-4463 was largely ambiguous. In the current study, we aimed to explore the function of miR-4463 in hypoxia-indu...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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IMR Press
2022-06-01
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| Series: | Frontiers in Bioscience-Landmark |
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| Online Access: | https://www.imrpress.com/journal/FBL/27/6/10.31083/j.fbl2706175 |
| _version_ | 1811236452446502912 |
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| author | Xuemei He Fei Wu Yang Zhou Jian Wu Yang Long Xiangyu Zhou |
| author_facet | Xuemei He Fei Wu Yang Zhou Jian Wu Yang Long Xiangyu Zhou |
| author_sort | Xuemei He |
| collection | DOAJ |
| description | Background: Our previous study revealed aberrant miR-4463 expression in the vascular tissues of patients with arteriosclerosis obliterans of the lower extremities (ASO), but the role of miR-4463 was largely ambiguous. In the current study, we aimed to explore the function of miR-4463 in hypoxia-induced endothelial cells and determine its molecular mechanisms. Methods: CCK-8 assay and flow cytometry were performed to evaluate cell viability and apoptosis. Adenovirus carrying mRFP-GFP-LC3 was employed to monitor cellular autophagy, and mitochondrial membrane potential was determined by JC-1 staining. Moreover, dual-luciferase reporter gene assay, qPCR, western blot and siRNA analysis were carried out to explore the potential molecular mechanisms. Results: Hypoxia significantly elevated the miR-4463 expression in primary human umbilical vein endothelial cells (HUVEC). Overexpression of miR-4463 inhibited hypoxia-induced autophagy by suppressing the formation of autophagosomes and autolysosomes, resulting in reduced cell viability and increased apoptosis, and these effects were reversed by miR-4463 inhibitor. Furthermore, activation of autophagy induced by miR-4463 inhibitor attenuated HUVECs apoptosis in hypoxic conditions. Mechanically, the results of the dual-luciferase reporter gene assay discovered that miR-4463 directly targeted Unc-51 like kinase 1 (ULK1). The silence of ULK1 blocked miR-4463 inhibitor-activated autophagy and further facilitated apoptosis under hypoxic conditions. Conclusions: Our findings indicate that miR-4463 is an essential regulator of hypoxia-induced autophagy and apoptosis in endothelial cells via directly targeting ULK1. Inhibition of miR-4463 might be a potential strategy to protect endothelial cells and maintain vascular function in patients with lower limb ischemia and its complications. |
| first_indexed | 2024-04-12T12:09:01Z |
| format | Article |
| id | doaj.art-6102d5cbcb224da890487d566053b4b3 |
| institution | Directory Open Access Journal |
| issn | 2768-6701 |
| language | English |
| last_indexed | 2024-04-12T12:09:01Z |
| publishDate | 2022-06-01 |
| publisher | IMR Press |
| record_format | Article |
| series | Frontiers in Bioscience-Landmark |
| spelling | doaj.art-6102d5cbcb224da890487d566053b4b32022-12-22T03:33:38ZengIMR PressFrontiers in Bioscience-Landmark2768-67012022-06-0127617510.31083/j.fbl2706175S2768-6701(22)00534-2miR-4463 Regulates Hypoxia-Induced Autophagy and Apoptosis by Targeting ULK1 in Endothelial CellsXuemei He0Fei Wu1Yang Zhou2Jian Wu3Yang Long4Xiangyu Zhou5Experimental Medicine Center, the Affiliated Hospital of Southwest Medical University, 646000 Luzhou, Sichuan, ChinaDepartment of Thyroid and Vascular Surgery, the Affiliated Hospital of Southwest Medical University, 646000 Luzhou, Sichuan, ChinaDepartment of Thyroid and Vascular Surgery, the Affiliated Hospital of Southwest Medical University, 646000 Luzhou, Sichuan, ChinaExperimental Medicine Center, the Affiliated Hospital of Southwest Medical University, 646000 Luzhou, Sichuan, ChinaExperimental Medicine Center, the Affiliated Hospital of Southwest Medical University, 646000 Luzhou, Sichuan, ChinaDepartment of Thyroid and Vascular Surgery, the Affiliated Hospital of Southwest Medical University, 646000 Luzhou, Sichuan, ChinaBackground: Our previous study revealed aberrant miR-4463 expression in the vascular tissues of patients with arteriosclerosis obliterans of the lower extremities (ASO), but the role of miR-4463 was largely ambiguous. In the current study, we aimed to explore the function of miR-4463 in hypoxia-induced endothelial cells and determine its molecular mechanisms. Methods: CCK-8 assay and flow cytometry were performed to evaluate cell viability and apoptosis. Adenovirus carrying mRFP-GFP-LC3 was employed to monitor cellular autophagy, and mitochondrial membrane potential was determined by JC-1 staining. Moreover, dual-luciferase reporter gene assay, qPCR, western blot and siRNA analysis were carried out to explore the potential molecular mechanisms. Results: Hypoxia significantly elevated the miR-4463 expression in primary human umbilical vein endothelial cells (HUVEC). Overexpression of miR-4463 inhibited hypoxia-induced autophagy by suppressing the formation of autophagosomes and autolysosomes, resulting in reduced cell viability and increased apoptosis, and these effects were reversed by miR-4463 inhibitor. Furthermore, activation of autophagy induced by miR-4463 inhibitor attenuated HUVECs apoptosis in hypoxic conditions. Mechanically, the results of the dual-luciferase reporter gene assay discovered that miR-4463 directly targeted Unc-51 like kinase 1 (ULK1). The silence of ULK1 blocked miR-4463 inhibitor-activated autophagy and further facilitated apoptosis under hypoxic conditions. Conclusions: Our findings indicate that miR-4463 is an essential regulator of hypoxia-induced autophagy and apoptosis in endothelial cells via directly targeting ULK1. Inhibition of miR-4463 might be a potential strategy to protect endothelial cells and maintain vascular function in patients with lower limb ischemia and its complications.https://www.imrpress.com/journal/FBL/27/6/10.31083/j.fbl2706175mir-4463autophagosomesautolysosomesapoptosis |
| spellingShingle | Xuemei He Fei Wu Yang Zhou Jian Wu Yang Long Xiangyu Zhou miR-4463 Regulates Hypoxia-Induced Autophagy and Apoptosis by Targeting ULK1 in Endothelial Cells Frontiers in Bioscience-Landmark mir-4463 autophagosomes autolysosomes apoptosis |
| title | miR-4463 Regulates Hypoxia-Induced Autophagy and Apoptosis by Targeting ULK1 in Endothelial Cells |
| title_full | miR-4463 Regulates Hypoxia-Induced Autophagy and Apoptosis by Targeting ULK1 in Endothelial Cells |
| title_fullStr | miR-4463 Regulates Hypoxia-Induced Autophagy and Apoptosis by Targeting ULK1 in Endothelial Cells |
| title_full_unstemmed | miR-4463 Regulates Hypoxia-Induced Autophagy and Apoptosis by Targeting ULK1 in Endothelial Cells |
| title_short | miR-4463 Regulates Hypoxia-Induced Autophagy and Apoptosis by Targeting ULK1 in Endothelial Cells |
| title_sort | mir 4463 regulates hypoxia induced autophagy and apoptosis by targeting ulk1 in endothelial cells |
| topic | mir-4463 autophagosomes autolysosomes apoptosis |
| url | https://www.imrpress.com/journal/FBL/27/6/10.31083/j.fbl2706175 |
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