LINC00943 acts as miR-338-3p sponge to promote MPP(+)-induced SK-N-SH cell injury by directly targeting SP1 in Parkinson's disease.

Abnormal expression of long non-coding RNA (lncRNA) is associated with the progression of Parkinson's disease (PD). LINC00943 has been proved to play an important role in the development of PD, so its role and mechanism in PD progression are worth further exploration. MPTP was used to construct PD mice model, and its active ingredient MPP+ was used to construct PD cell model. Cell proliferation and apoptosis were determined by MTT assay, EdU staining and flow cytometry. The protein levels of Cyclin D1, Bcl-2 and specificity protein 1 (SP1) were tested by western blot analysis. The concentrations of inflammation factors were examined by ELISA assay. The expression levels of LINC00943, microRNA (miR)-338-3p and SP1 were measured using quantitative real-time PCR. The interaction between miR-338-3p and LINC00943 or SP1 was confirmed using dual-luciferase reporter assay and RIP assay. Our data showed that LINC00943 was highly expressed in the brain tissues of MPTP-treated mice and MPP+-induced SK-N-SH cells. Knockdown of LINC00943 could promote the proliferation, while inhibit the apoptosis and inflammation of MPP+-induced SK-N-SH cells to alleviate cell injury. In terms of mechanism, we pointed out that LINC00943 could sponge miR-338-3p, and miR-338-3p could target SP1. The negative regulation of si-LINC00943 on MPP+-induced SK-N-SH cell injury could be reversed by miR-338-3p inhibitor. Moreover, miR-338-3p had a protective effect on SK-N-SH cells from MPP+-induced injury, which could be reversed by SP1 overexpression. Additionally, we confirmed that LINC00943 positively regulated SP1 via sponging miR-338-3p. To sum up, our data revealed that knockdown LINC00943 might alleviate PD progression through regulating the miR-338-3p/SP1 axis.

Sun X ，Zhang C ，Tao H ，Yao S ，Wu X ... -  《-》

Berberine Attenuates MPP(+)-Induced Neuronal Injury by Regulating LINC00943/miR-142-5p/KPNA4/NF-κB Pathway in SK-N-SH Cells.

Berberine plays a neuro-protective role in neurodegenerative diseases, including Parkinson's disease (PD). Long non-coding RNAs (lncRNAs) play critical roles in PD pathogenesis. The purpose of this study was to investigate whether LINC00943 was involved in the role of berberine in PD. 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) or 1-methyl-4-phenyl pyridine (MPP+) were used to construct PD mouse and cell models, respectively. Cell proliferation was evaluated by Cell Counting Kit-8 (CCK-8) and 5-Ethynyl-2'-deoxyuridine (Edu) assays. Inflammation and cell apoptosis were assessed by enzyme-linked immunosorbent assay (ELISA) and flow cytometry, respectively. Quantitative real-time PCR (qRT-PCR) was employed to test the expression of LINC00943, microRNA (miR)-142-5p, and karyopherin subunit alpha 4 (KPNA4) mRNA. The protein levels of NF-κB pathway-related markers and KPNA4 were measured by western blot. Oxidative stress level was assessed by corresponding kits. The interaction between miR-142-5p and LINC00943 or KPNA4 was determined via dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Berberine inhibited MPP+-induced injury in SK-N-SH cells by promoting cell proliferation and suppressing inflammation, apoptosis, and oxidative injury. LINC00943 and KPNA4 were upregulated and miR-142-5p was downregulated in PD mouse and cell models. LINC00943 (or KPNA4) overexpression or miR-142-5p inhibition abated the neuro-protective role of berberine in PD cell model. Moreover, miR-142-5p was a target of LINC00943, and KPNA4 could specially bind to miR-142-5p. Additionally, berberine inhibited NF-κB pathway by regulating LINC00943/miR-142-5p/KPNA4 axis. Berberine protected SK-N-SH cell from MPP+-induced neuronal damage via regulating LINC00943/miR-142-5p/KPNA4/NF-κB pathway, highlighting novel evidence for the neuro-protective role of berberine in PD.

Li X ，Su Y ，Li N ，Zhang FR ，Zhang N ... -  《-》

LINC00943 knockdown exerts neuroprotective effects in Parkinson's disease through regulates CXCL12 expression by sponging miR-7-5p.

Lian H ，Wang B ，Lu Q ，Chen B ，Yang H ... -  《-》

A novel LINC00943/miR-671-5p/ELAVL1 ceRNA crosstalk regulates MPP(+) toxicity in SK-N-SH cells.

The competing endogenous RNA (ceRNA) activity of long non-coding RNAs (lncRNAs) has profound effects in pathological disorders, including Parkinson's disease. Here, we focused on the LINC00943-mediated ceRNA network for the regulation of LINC00943 in MPP+ toxicity in SK-N-SH cells. SK-N-SH cells were exposed to 1-methyl-4-phenylpyridinium (MPP+). LINC00943, miR-671-5p and ELAV like RNA binding protein 1 (ELAVL1) were quantified by real-time quantitative PCR (RT-qPCR) or western blot. Cell viability and apoptosis were gauged by Cell Counting Kit-8 (CCK-8) assay and flow cytometry, respectively. Direct relationship between miR-671-5p and LINC00943 or ELAVL1 was confirmed by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Our data validated that LINC00943 regulated MPP+-evoked injury in SK-N-SH cells. LINC00943 regulated miR-671-5p expression by binding to miR-671-5p. Moreover, miR-671-5p was identified as a molecular mediator of LINC00943 in regulating SK-N-SH cell injury induced by MPP+. MiR-671-5p targeted and inhibited ELAVL1, and miR-671-5p-mediated inhibition of ELAVL1 impacted MPP+-evoked SK-N-SH cell injury. Furthermore, LINC00943 involved the post-transcriptional regulation of ELAVL1 through miR-671-5p competition. Our present study has established a novel mechanism, the LINC00943/miR-671-5p/ELAVL1 ceRNA crosstalk, for the regulation of LINC00943 on MPP+ toxicity in SK-N-SH cells.

Zhang X ，Luan N ，Shi J 《-》

SOS1-IT1 silencing alleviates MPP(+)-induced neuronal cell injury through regulating the miR-124-3p/PTEN/AKT/mTOR pathway.

Long non-coding RNA (lncRNA) has been found to be involved in the regulation of a variety of disease progression, including Parkinson's disease (PD). However, the role and underlying mechanism of SOS1 intronic transcript 1 (SOS1-IT1) in the progression of PD is still unclear. 1-methyl-4-phenyl pyridine (MPP+) induced SK-N-SH cells were used to construct PD cell models in vitro. The expression levels of SOS1-IT1, microRNA (miR)-124-3p and phosphatase and tensin homolog (PTEN) were determined using quantitative real-time PCR. Cell counting kit 8 assay and flow cytometry were used to measure cell viability and apoptosis. Western blot analysis was performed to detect protein expression. The levels of inflammation cytokines and oxidative stress markers were examined to assess cell inflammation and oxidative stress. In addition, dual-luciferase reporter assay, RIP assay and RNA pull-down assay were used to confirm RNA interaction. Our results showed that SOS1-IT1 was upregulated in MPP+-induced SK-N-SH cells, and its silencing reversed the inhibition effect of MPP+ on the viability and the promotion effect on the apoptosis, inflammation and oxidative stress of SK-N-SH cells. MiR-124-3p was targeted by SOS1-IT1, and its inhibitor reversed the suppressive effect of SOS1-IT1 knockdown on MPP+-induced SK-N-SH cell injury. Furthermore, PTEN was a target of miR-124-3p, and the reduction effect of miR-124-3p on MPP+-induced SK-N-SH cell injury was reversed by PTEN overexpression. Additionally, the activity of AKT/mTOR pathway was regulated by the SOS1-IT1/miR-124-3p/PTEN axis. In conclusion, SOS1-IT1 regulated the miR-124-3p/PTEN/AKT/mTOR pathway to participate in the regulation of MPP+-induced neuronal cell injury, indicating the SOS1-IT1 might be an effective therapeutic target for PD.

Fan J ，Wu D ，Guo Y ，Yang Z ... -  《-》