MicroRNA-21-5p Reduces Hypoxia/Reoxygenation-Induced Neuronal Cell Damage through Negative Regulation of CPEB3
Objectives. To explore the role of microRNA-21-5p (miR-21-5p) in hypoxia/reoxygenation- (H/R-) induced HT22 cell damage. Methods. The hypoxia/reoxygenation (H/R) model was established in mouse neuronal cells HT22. Cell Counting Kit-8 (CCK-8) and qRT-PCR were used to determine the effects of H/R trea...
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| Format: | Article |
| Language: | English |
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Wiley
2021-01-01
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| Series: | Analytical Cellular Pathology |
| Online Access: | http://dx.doi.org/10.1155/2021/5543212 |
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| author | Bin Wang Ping Yu Wei Lin Zhaohui Zhai |
| author_facet | Bin Wang Ping Yu Wei Lin Zhaohui Zhai |
| author_sort | Bin Wang |
| collection | DOAJ |
| description | Objectives. To explore the role of microRNA-21-5p (miR-21-5p) in hypoxia/reoxygenation- (H/R-) induced HT22 cell damage. Methods. The hypoxia/reoxygenation (H/R) model was established in mouse neuronal cells HT22. Cell Counting Kit-8 (CCK-8) and qRT-PCR were used to determine the effects of H/R treatment on cell viability and miR-21-5p expression. HT22 cells were transfected with miR-21-5p mimic or negative control (NC) followed by the induction of H/R; cell viability, apoptosis, and SOD, MDA, and LDH activities were detected. Besides, the apoptosis-related proteins including BAX, BCL2, cleaved caspase-3, and caspase-3 as well as proteins of EGFR/PI3K/AKT signaling pathways were measured by Western blot. To verify the target relation between cytoplasmic polyadenylation element binding protein 3 (CPEB3) and miR-21-5p, luciferase reporter gene experiment was performed. After cotransfection with miR-21-5p mimic and CPEB3 plasmids, the reversal effects of CPEB3 on miR-21-5p in H/R damage were studied. Results. H/R treatment could significantly reduce the cell viability (P<0.05) and miR-21-5p levels (P<0.05) in HT22 cells. After overexpressing miR-21-5p, cell viability was increased (P<0.05) under H/R treatment, and the apoptosis rate and the levels of apoptosis-related proteins were suppressed (all P<0.05). Furthermore, SOD activity was increased (P<0.05), while MDA and LDH activity was decreased (both P<0.05). Besides, miR-21-5p could restore the activation of the EGFR/PI3K/AKT signaling pathway inhibited by H/R treatment (all P<0.05). The luciferase reporter gene experiment verified that CPEB3 is the target of miR-21-5p (P<0.05). When coexpressing miR-21-5p mimic and CPEB3 in the cells, the protective effects of miR-21-5p under H/R were reversed (all P<0.05), and the activation of the EGFR/PI3K/AKT pathway was also inhibited (all P<0.05). Conclusion. This study showed that miR-21-5p may regulate the EGFR/PI3K/AKT signaling pathway by targeting CPEB3 to reduce H/R-induced cell damage and apoptosis. |
| format | Article |
| id | doaj-art-6475b3c5d1ec4bed8453bf1692b0730f |
| institution | Kabale University |
| issn | 2210-7185 |
| language | English |
| publishDate | 2021-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Analytical Cellular Pathology |
| spelling | doaj-art-6475b3c5d1ec4bed8453bf1692b0730f2025-02-03T01:07:07ZengWileyAnalytical Cellular Pathology2210-71852021-01-01202110.1155/2021/5543212MicroRNA-21-5p Reduces Hypoxia/Reoxygenation-Induced Neuronal Cell Damage through Negative Regulation of CPEB3Bin Wang0Ping Yu1Wei Lin2Zhaohui Zhai3Department of NeurologyDepartment of NeurologyPain DepartmentDepartment of Plastic SurgeryObjectives. To explore the role of microRNA-21-5p (miR-21-5p) in hypoxia/reoxygenation- (H/R-) induced HT22 cell damage. Methods. The hypoxia/reoxygenation (H/R) model was established in mouse neuronal cells HT22. Cell Counting Kit-8 (CCK-8) and qRT-PCR were used to determine the effects of H/R treatment on cell viability and miR-21-5p expression. HT22 cells were transfected with miR-21-5p mimic or negative control (NC) followed by the induction of H/R; cell viability, apoptosis, and SOD, MDA, and LDH activities were detected. Besides, the apoptosis-related proteins including BAX, BCL2, cleaved caspase-3, and caspase-3 as well as proteins of EGFR/PI3K/AKT signaling pathways were measured by Western blot. To verify the target relation between cytoplasmic polyadenylation element binding protein 3 (CPEB3) and miR-21-5p, luciferase reporter gene experiment was performed. After cotransfection with miR-21-5p mimic and CPEB3 plasmids, the reversal effects of CPEB3 on miR-21-5p in H/R damage were studied. Results. H/R treatment could significantly reduce the cell viability (P<0.05) and miR-21-5p levels (P<0.05) in HT22 cells. After overexpressing miR-21-5p, cell viability was increased (P<0.05) under H/R treatment, and the apoptosis rate and the levels of apoptosis-related proteins were suppressed (all P<0.05). Furthermore, SOD activity was increased (P<0.05), while MDA and LDH activity was decreased (both P<0.05). Besides, miR-21-5p could restore the activation of the EGFR/PI3K/AKT signaling pathway inhibited by H/R treatment (all P<0.05). The luciferase reporter gene experiment verified that CPEB3 is the target of miR-21-5p (P<0.05). When coexpressing miR-21-5p mimic and CPEB3 in the cells, the protective effects of miR-21-5p under H/R were reversed (all P<0.05), and the activation of the EGFR/PI3K/AKT pathway was also inhibited (all P<0.05). Conclusion. This study showed that miR-21-5p may regulate the EGFR/PI3K/AKT signaling pathway by targeting CPEB3 to reduce H/R-induced cell damage and apoptosis.http://dx.doi.org/10.1155/2021/5543212 |
| spellingShingle | Bin Wang Ping Yu Wei Lin Zhaohui Zhai MicroRNA-21-5p Reduces Hypoxia/Reoxygenation-Induced Neuronal Cell Damage through Negative Regulation of CPEB3 Analytical Cellular Pathology |
| title | MicroRNA-21-5p Reduces Hypoxia/Reoxygenation-Induced Neuronal Cell Damage through Negative Regulation of CPEB3 |
| title_full | MicroRNA-21-5p Reduces Hypoxia/Reoxygenation-Induced Neuronal Cell Damage through Negative Regulation of CPEB3 |
| title_fullStr | MicroRNA-21-5p Reduces Hypoxia/Reoxygenation-Induced Neuronal Cell Damage through Negative Regulation of CPEB3 |
| title_full_unstemmed | MicroRNA-21-5p Reduces Hypoxia/Reoxygenation-Induced Neuronal Cell Damage through Negative Regulation of CPEB3 |
| title_short | MicroRNA-21-5p Reduces Hypoxia/Reoxygenation-Induced Neuronal Cell Damage through Negative Regulation of CPEB3 |
| title_sort | microrna 21 5p reduces hypoxia reoxygenation induced neuronal cell damage through negative regulation of cpeb3 |
| url | http://dx.doi.org/10.1155/2021/5543212 |
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