Ruiguang Liu1, Fenlin Li2, Weijie Zhao3. 1. Department of Neurology, Changle People's Hospital, Weifang, Shandong, China. 2. Department of Geriatrics, Chengyang People's Hospital, Qingdao, Shandong, China. 3. Department of Geriatrics, Changle People's Hospital, Weifang, Shandong, China. Electronic address: v0jov0@163.com.
Abstract
BACKGROUND: Some long non-coding RNAs (lncRNAs) have been suggested to play critical roles in Parkinson's disease (PD) pathogenesis, including nuclear enriched abundant transcript 1 (NEAT1). The purpose of this study was to further elucidate the molecular mechanism of NEAT1 in PD. METHODS: The expression levels of NEAT1, miR-212-5p and RAB3A-interacting protein (RAB3IP) were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability and apoptosis were evaluated by Cell Counting Kit-8 (CCK-8) assay and flow cytometry analysis, respectively. Western blot analysis was applied to detect the protein expression of IL-1β, TNF-α and RAB3IP. The LDH activity, ROS generation and SOD activity were measured by Lactate LDH activity assay kit, ROS assay kit, and SOD activity assay kit, respectively. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to verify the relationship between miR-212-5p and NEAT1 or mRNA of RAB3IP. 1-methyl-4-phenylpyridinium ion (MPP+)-treated SK-N-SH cells were used as an in vitro model of PD. RESULTS: NEAT1 and RAB3IP were upregulated while miR-212-5p was downregulated in SK-N-SH cells treated with MPP+. NEAT1 knockdown or miR-212-5p overexpression inhibited MPP+-induced apoptosis, inflammation and cytotoxicity in SK-N-SH cells. Moreover, miR-212-5p was a direct target of NEAT1 and its downregulation reversed the eff ;ects caused by NEAT1 knockdown in MPP+-induced SK-N-SH cells. Furthermore, RAB3IP was a downstream target of miR-212-5p and its overexpression attenuated the effects of miR-212-5p restoration in MPP+-induced SK-N-SH cells. Besides, NEAT1 acted as a molecular sponge of miR-212-5p to regulate RAB3IP expression. CONCLUSION: NEAT1 knockdown suppressed MPP+-induced apoptosis, inflammation and cytotoxicity in SK-N-SH cells through regulating miR-212-5p and RAB3IP expression, providing a possible therapeutic strategy for PD patients.
BACKGROUND: Some long non-coding RNAs (lncRNAs) have been suggested to play critical roles in Parkinson's disease (PD) pathogenesis, including nuclear enriched abundant transcript 1 (NEAT1). The purpose of this study was to further elucidate the molecular mechanism of NEAT1 in PD. METHODS: The expression levels of NEAT1, miR-212-5p and RAB3A-interacting protein (RAB3IP) were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability and apoptosis were evaluated by Cell Counting Kit-8 (CCK-8) assay and flow cytometry analysis, respectively. Western blot analysis was applied to detect the protein expression of IL-1β, TNF-α and RAB3IP. The LDH activity, ROS generation and SOD activity were measured by Lactate LDH activity assay kit, ROS assay kit, and SOD activity assay kit, respectively. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to verify the relationship between miR-212-5p and NEAT1 or mRNA of RAB3IP. 1-methyl-4-phenylpyridinium ion (MPP+)-treated SK-N-SH cells were used as an in vitro model of PD. RESULTS:NEAT1 and RAB3IP were upregulated while miR-212-5p was downregulated in SK-N-SH cells treated with MPP+. NEAT1 knockdown or miR-212-5p overexpression inhibited MPP+-induced apoptosis, inflammation and cytotoxicity in SK-N-SH cells. Moreover, miR-212-5p was a direct target of NEAT1 and its downregulation reversed the eff ;ects caused by NEAT1 knockdown in MPP+-induced SK-N-SH cells. Furthermore, RAB3IP was a downstream target of miR-212-5p and its overexpression attenuated the effects of miR-212-5p restoration in MPP+-induced SK-N-SH cells. Besides, NEAT1 acted as a molecular sponge of miR-212-5p to regulate RAB3IP expression. CONCLUSION:NEAT1 knockdown suppressed MPP+-induced apoptosis, inflammation and cytotoxicity in SK-N-SH cells through regulating miR-212-5p and RAB3IP expression, providing a possible therapeutic strategy for PDpatients.
Authors: Shubhra Acharya; Antonio Salgado-Somoza; Francesca Maria Stefanizzi; Andrew I Lumley; Lu Zhang; Enrico Glaab; Patrick May; Yvan Devaux Journal: Int J Mol Sci Date: 2020-09-06 Impact factor: 5.923