Research reportDeficiency of NEAT1 prevented MPP+-induced inflammatory response, oxidative stress and apoptosis in dopaminergic SK-N-SH neuroblastoma cells via miR-1277-5p/ARHGAP26 axis
Introduction
Parkinson’s disease (PD), the second most common neurodegenerative disorder after alzheimer's disease (Poewe et al., 2017). The incidence of PD is rapidly increased with age in China (Li et al., 2019), making PD as one main threat in aged people. Although the etiology of PD is still unclear, genetic factors and environmental factors (such as toxicants) have been declared to evoke the development of PD (Dunn et al., 2019). The common etiopathogenesis of PD is ascribed to the loss of dopaminergic neurons in substantia nigra pars compacta, and the accumulation of Lewy bodies in the brain (Poewe et al., 2017). Extensive evidences have indicated that inflammatory damage and oxidative injury are responsible to neurodegeneration and neuroinflammation in PD (Anderson et al., 2018, Puspita et al., 2017). N-methyl-4-phenylpyridinium (MPP+) is the active metabolite of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a kind of neurotoxin that selectively destroys dopaminergic neurons, eventually leading to the occurrence of PD (Jiang et al., 2019b). Therefore, clarifying the mechanism of MPP+-induced PD is important to clarify the pathogenesis of PD.
Noncoding RNAs including long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) are abnormally expressed in neurodevelopment and neurodegenerative diseases including PD (Majidinia et al., 2016, Oe et al., 2019). Growing evidences support a key role of lncRNAs and miRNAs in dopaminergic dysfunction in neurological pathologies, such as PD (Carrick et al., 2016). LncRNAs are longer than 200 nucleotides (nt), and are structurally similar to message RNAs (mRNAs), even though lncRNAs have little protein coding capacity (Lyu et al., 2019). Newest researches on the pathogenesis of diseases have elucidated that lncRNAs are emerging targets for diagnosis and drug treatment of PD (Lyu et al., 2019), and that the role of several lncRNAs have already been illuminated in the pathogenesis of PD (Fan et al., 2019, Majidinia et al., 2016). However, the research on contribution of lncRNAs in PD remains infant. LncRNA nuclear enriched assembly transcript 1 (NEAT1) is aberrantly-mostly upregulated in non-cancerous pathological conditions, including neurodegenerative diseases (Prinz et al., 2019). Furthermore, the de novo investigations have disclosed the close relationship among NEAT1, inflammatory response (Xie et al., 2019, Zhang et al., 2019b) and oxidative stress (Simchovitz et al., 2019, Sunwoo et al., 2017). Here, we intended to explore the role of NEAT1 in inflammatory injury and oxidative damage in PD.
MiRNAs are a class of transcripts (19–24 nt) that control the expression of multiple mRNAs at post-transcriptional level (Majidinia et al., 2016). Numerous documents have expounded the prominent role of miRNAs in PD (Goh et al., 2019), and cell-free miRNAs are considered as potential biomarkers for central nervous system disorders including PD (Doxakis, 2020, van den Berg et al., 2020). MiRNA (miR)-1277-5p is associated with accurate molecular classification of different cancers, such as kidney cancer, colorectal cancer, hepatocellular carcinoma (Cao et al., 2019, Motieghader et al., 2017, Youssef et al., 2011). Nevertheless, the function of miR-1277-5p is not well understood presently. Thus, we wondered the role of miR-1277-5p in MPP+-induced dopaminergic neurons.
The presented study focused on the expression of NEAT1 and miR-1277-5p in MPP+-induced model of PD in dopaminergic neuroblastoma SK-N-SH cells. Furthermore, the role of both was also investigated in inflammatory injury and oxidative damage in response to MPP+. Notably, the molecular mechanism of NEAT1 was further clarified through functioning as a competing endogenous RNA (ceRNA) of miR-1277-5p to regulate Rho GTPase-activating protein 26 (ARHGAP26, also known as GRAF), a negative regulator of the Rho family (Wang et al., 2013).
Section snippets
Expression of NEAT1 was upregulated in cell model of PD
An in vitro model of PD was established in dopaminergic neuroblastoma SK-N-SH cells treated with MPP+. As shown in Fig. 1A and 1B, expression level of NEAT1 was gradually increased in SK-N-SH cells after treatment of 0.25–1 mM MPP+ for 24 h and 1 mM MPP+ for 12–48 h. Thus, we considered that NEAT1 was upregulated in MPP+-induced SK-N-SH cells in a certain of concentration- and time-dependent manner, suggesting a potential role of NEAT1 in neuronal cell injury in PD. And, 1 mM MPP+ treatment for
Discussion
In this study, we proposed that NEAT1 might contribute to dopaminergic neuroblastoma SK-N-SH cell injury by regulating miR-1277-5p and ARHGAP26 to modulate cell viability, apoptosis, inflammation and oxidative stresses, suggesting aNEAT1-miR-1277-5p-ARHGAP26 ceRNA pathway underlying the pathogenesis of PD (Fig. 7).
NEAT1 was overexpressed in both PD cell model and drug-inducible neuroprotection against oxidative damage (Simchovitz et al., 2019). This pointed out a complicated involvement of
Cell model of PD
The dopaminergic neuroblastoma cell lines, SK-N-SH (no. 86012802) and SH-SY5Y (no. 94030304; differentiated from SK-N-SH) were from European Collection of Authenticated Cell Cultures (Public Health England, UK), and MPP+ (no. D048) was from Sigma-Aldrich (St. Louis, MO, USA). For establishment of cell model of PD, SK-N-SH cells were treated with 0.25, 0.5 and 1 mM of MPP+ for 0–48 h in Dulbecco’s modified Eagle’s medium (Hyclone, Logan, Utah, USA) plus 10% fetal bovine serum (Hyclone) in a
CRediT authorship contribution statement
Shufang Zhou: Validation, Investigation, Writing - original draft, Writing - review & editing, Project administration. Dan Zhang: Conceptualization, Methodology, Writing - original draft, Writing - review & editing. Junnan Guo: Conceptualization, Methodology, Writing - original draft, Writing - review & editing. Zhenzhen Chen: Formal analysis, Data curation, Software. Yong Chen: Formal analysis, Data curation, Validation, Investigation. Junshi Zhang: Validation, Investigation.
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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