Hsa_circ_0002468 Regulates the Neuronal Differentiation of SH-SY5Y Cells by Modulating the MiR-561/E2F8 Axis

Abstract

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/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.

RETRACTED:CircFBXW7alleviates glioma progression through regulatingmiR‐23a‐3p/PTENaxis

“…Along with the development of gene sequencing, microarray, and genomics, an increasing number of circRNAs have been found to be involved in pathophysiological processes of the central nervous system, which also play an important role in the development of diseases in the central nervous system (D'Ambra, Capauto, & Morlando, 2019; He et al, 2019; Zhang, Yu, Bao, & Sun, 2019). Moreover, the abnormal expression of circRNAs may lead to a series of diseases of the central nervous system, which have a large effect on neuron cell differentiation (Yang, Xiang, et al, 2019). Currently, the important role of circRNAs in the pathological process of glioma is widely accepted (Nie, Li, Zhang, & Liu, 2019).…”

Toward Spontaneous Neuronal Differentiation of SH-SY5Y Cells Using Novel Three-Dimensional Electropolymerized Conductive Scaffolds

“…To induce differentiation to neurons, SH-SY5Y cells are usually cultured in the presence of retinoic acid alone or combined with other chemicals or neurotrophic factors. [1][2][3][4][5][6] Neurotrophic factors, such as brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF), play an important role in neuronal regeneration, but their clinical use is limited since they are not able to cross the blood brain barrier. 7 In the recent years, the synergy between nanotechnology and biomedicine in therapeutic and diagnostic tools has grown significantly.…”

Circular RNAs: The Novel Actors in Pathophysiology of Spinal Cord Injury

“…The circRNAs that are widely expressed in the mammalian brain (Rybak-Wolf et al, 2015 ; Hanan et al, 2017 ) have been suggested to participate in neurodevelopment (Hansen et al, 2013 ; Memczak et al, 2013 ), neuronal proliferation, and differentiation (Chen and Schuman, 2016 ; Yang et al, 2019 ; Suenkel et al, 2020 ). For example, a significant increase of certain circRNAs has been associated with myelination and neural maturation in the brain and spinal cord during postnatal development (Di Agostino et al, 2020 ), revealing the key role of circRNAs also in motoneuron development in cortex, brainstem, or spinal cord.…”

“…Another role of the circRNAs has been reported during development of the neocortex where they regulate spontaneous neuronal differentiation and are important to maintain the pool of cell progenitors (Suenkel et al, 2020 ). The spatiotemporal expression and function of lineage-specific circRNAs partly underlie the regulation of transcription factors to induce neuronal and glial differentiation and proliferation (Yang et al, 2019 ). Moreover, since circRNAs exhibit differential expression in different brain cells and areas, they could also represent a valuable tool for the development of novel targeted markers for gliomas progression and tumorigenicity (Sun et al, 2020 ).…”