Hsa_circ_0002468 Regulates the Neuronal Differentiation of SH-SY5Y Cells by Modulating the MiR-561/E2F8 Axis
| Autor: | Dan Yu, Ai Qun Liu, Guanghong Xiang, Songlin Yang, Gaoya Zhou, Guoshuai Yang, Minhui Yang, Weifeng He |
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| Rok vydání: | 2019 |
| Předmět: |
RNA
Untranslated SH-SY5Y E2F6 Transcription Factor Apoptosis 030204 cardiovascular system & hematology Real-Time Polymerase Chain Reaction Cell Line 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Lab/In Vitro Research Cell Movement Humans Binding site E2F Transcription factor Neuregulins Cell Proliferation Neurons Reporter gene Chemistry Cell growth Computational Biology Cell Differentiation RNA Circular General Medicine Deoxyuridine Cell biology Repressor Proteins MicroRNAs Cell culture 030220 oncology & carcinogenesis RNA |
| Zdroj: | Medical Science Monitor : International Medical Journal of Experimental and Clinical Research |
| ISSN: | 1643-3750 |
| Popis: | BACKGROUND It has been shown that circular RNAs (circRNAs) play a vital role in the regulation of neuronal differentiation; however, the precise role of circRNAs in human neuronal differentiation remains largely unexplored. MATERIAL AND METHODS A dual-luciferase reporter assay was carried out to confirm the targets of hsa_circ_0002468, miR-561, E2F8 (E2F transcription factor 8, a protein coding gene), and miR-561. We detected the expression of hsa_circ_0002468, miR-561, and E2F8 by using quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In addition, we performed the functional experiments by using a BrdU (5-bromo-2'-deoxyuridine) assay and qRT-PCR analyses. RESULTS In this study, we showed that hsa_circ_0002468 can act as a sponge of miR-561 to regulate SH-SY5Y proliferation and differentiation. A bioinformatics analysis showed that hsa_circ_0002468 had a binding site that corresponded to miR-561, which was verified by dual-luciferase reporter assay. The expression of hsa_circ_0002468 was increased during SH-SY5Y differentiation and was inversely correlated with miR-561 expression. Using qRT-PCR analysis, we showed that hsa_circ_0002468 negatively regulated miR-561 in SH-SY5Y cells. Intriguingly, the overexpression of hsa_circ_0002468 increased SH-SY5Y differentiation and reduced SH-SY5Y proliferation; the suppression of hsa_circ_0002468 led to decreased SH-SY5Y differentiation levels and increased SH-SY5Y proliferation levels. Additionally, overexpression of miR-561 rescued the SH-SY5Y proliferation deficiency induced by hsa_circ_0002468 overexpression and abolished the SH-SY5Y differentiation promoted by hsa_circ_0002468. Furthermore, E2F8 was validated as a direct target of miR-561. CONCLUSIONS Our data suggested that hsa_circ_0002468 was a novel circRNA that regulated SH-SY5Y cell proliferation and differentiation via targeting the miR-561/E2F8 axis. Therefore, manipulating hsa_circ_0002468 in SH-SY5Y cells could be a novel strategy to develop novel interventions for the treatment of relevant neurological disorders. |
| Databáze: | OpenAIRE |
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