Related Experiment Video
Updated: Oct 14, 2025
Molecular Modulation by Lentivirus-Delivered Specific shRNAs in Endoplasmic Reticulum Stressed Neurons
Published on: April 24, 2021
Circular RNA circ_0070441 regulates MPP+-triggered neurotoxic effect in SH-SY5Y cells via miR-626/IRS2 axis
Xuqing Cao1, Jiangtao Guo2, Hideki Mochizuki3
1Department of Neurology, People's Hospital of Ningxia Hui Autonomous Region, Zhengyuan North Street, Jinfeng District, Yinchuan, 750002, China. fdnzweb@163.com.
Abstract:
Circular RNAs (circRNAs) was suggested to play crucial regulatory roles in various human diseases, including Parkinson's disease (PD). This research aimed to investigate the function and potential mechanism of circ_0070441 in PD. MPP+ (1-methyl-4-phenylpyridinium)-treated SH-SY5Y cells was used as an in vitro cellular PD model. The expressions of circ_0070441, microRNA (miR)-626 and insulin receptor substrate 2 (IRS2) were measured by quantitative real-time polymerase chain reaction (RT-qPCR) or western blot. Cell Counting Kit-8 (CCK-8) assay, Cytotoxicity Detection Kit (Lactate Dehydrogenase), flow cytometry and Caspase-3 Assay Kit were used to detect cell viability, LDH release, cell apoptosis and caspase-3 activity, respectively. The levels of inflammation-related factors were detected by enzyme-linked immunosorbent assay (ELISA). The correlation among circ_0070441, miR-626 and IRS2 were confirmed by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull-down assay. The levels of circ_0070441 and IRS2 were increased while miR-626 expression was decreased in MPP+-treated SH-SY5Y cells in dose- and time-dependent manners. Depletion of circ_0070441 alleviated MPP+-triggered neuronal damage by regulating cell apoptosis and inflammation. Circ_0070441 acted as a sponge for miR-626, and IRS2 was a target of miR-626. Besides, the neuroprotective effects of circ_0070441 knockdown or miR-626 overexpression were partly overturned by the suppression of miR-626 or IRS2 overexpression. Moreover, circ_0070441 upregulated IRS2 expression by interacting with miR-626. In summary, circ_0070441 aggravated MPP+-triggered neurotoxic effect in SH-SY5Y cells by regulating miR-626/IRS2 axis.
Insights
Circular RNAs (circRNAs) worsen Parkinson's disease (PD) neurotoxicity by upregulating IRS2 via sponging miR-626. Reducing circ_0070441 protects against MPP+-induced neuronal damage in a cellular PD model.
Area of Science:
- Molecular Biology
- Neuroscience
- Genetics
Background:
- Circular RNAs (circRNAs) are implicated in human diseases, including Parkinson's disease (PD).
- Understanding the role of specific circRNAs, like circ_0070441, in PD pathogenesis is crucial for developing therapeutic strategies.
- The MPP+ (1-methyl-4-phenylpyridinium) treated SH-SY5Y cell line serves as a widely accepted in vitro model for studying PD.
Purpose of the Study:
- To investigate the function and underlying mechanism of circ_0070441 in a cellular model of Parkinson's disease.
- To elucidate the regulatory relationship between circ_0070441, microRNA-626 (miR-626), and insulin receptor substrate 2 (IRS2) in the context of neurotoxicity.
Main Methods:
- Quantitative real-time PCR (RT-qPCR) and Western blot were used to measure gene and protein expression levels.
- Cell viability, apoptosis, and inflammation were assessed using CCK-8, LDH release, flow cytometry, Caspase-3 assays, and ELISA.
- Dual-luciferase reporter, RNA immunoprecipitation (RIP), and RNA pull-down assays were employed to confirm molecular interactions.
Main Results:
- MPP+ treatment increased circ_0070441 and IRS2 expression while decreasing miR-626 levels in SH-SY5Y cells in a dose- and time-dependent manner.
- Knockdown of circ_0070441 reduced MPP+-induced neuronal damage, apoptosis, and inflammation.
- circ_0070441 directly targets miR-626, and miR-626 targets IRS2, establishing a circ_0070441/miR-626/IRS2 regulatory axis.
Conclusions:
- circ_0070441 exacerbates MPP+-induced neurotoxicity in SH-SY5Y cells by upregulating IRS2 expression through sponging miR-626.
- The circ_0070441/miR-626/IRS2 pathway represents a potential therapeutic target for Parkinson's disease.
Related Concept Videos
02:15Experimental RNAi
01:22MicroRNAs
01:23RNA Interference
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
02:23RNA Editing
02:23Abnormal Proliferation
02:30siRNA - Small Interfering RNAs
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...