Nrf2-regulated miR-380-3p blocks the translation of Sp3 protein and its mediation of paraquat-induced toxicity in mouse neuroblastoma N2a cells
作者单位：Fujian Provincial Key Laboratory of Environmental Factors and Cancer School of Public HealthFujian Medical University The Key Laboratory of Environment and HealthSchool of Public Health Fujian Medical University Center for Drug Non-clinical Evaluation and Research of Guangdong Institute of Applied Bio-resources Department of Preventive Medicine School of Public Health Fujian Medical University Department of ManagementFujian Health College Department of Molecular PharmacologyAlbert Einstein College of Medicine Department of Environmental and Occupational Health Sciences University of Louisville Department of Epidemiology and Health Statistics School of Public Health Fujian Medical University
会议名称：《中国毒理学会第九次全国毒理学大会》
会议日期：2019年
学科分类：1002[医学-临床医学] 100201[医学-内科学(含：心血管病、血液病、呼吸系病、消化系病、内分泌与代谢病、肾病、风湿病、传染病)] 10[医学]
基 金：the National Natural Science Foundation in China(Grant 81573195, 81172715, 30800936) the Provincial Natural Science Foundation of Fujian province(2017 J01523) the Project of Health and Family Planning Commission of Fujian Province(2017-1-63) the Joint Funds for the innovation of science and Technology, Fujian province(2017Y9105) National Institute of Environmental Health Sciences, NIEHS,R01ES07331, R01ES10563 and R01ES020852
关 键 词：paraquat neurotoxicity Nrf2 miR-380-3p Sp3 p21 CaM
摘 要：Laboratorial and epidemiological research has established a relationship between paraquat（PQ） exposure and a risk for Parkinson′s disease. Previously, we have investigated the effects of nuclear factor erythroid 2 related factor 2（Nrf2） and microRNAs in PQ-induced neurotoxicity, addressing the function of miR-380-3p, a microRNA dysregulated by PQ, as well as Nrf2 deficiency. Nrf2 is known to mediate the expression of a variety of genes, including non-coding genes. By chromatin immunoprecipitation, we identified the relationship between Nrf2 and miR-380-3p in transcriptional regulation. qRT-PCR, western-blots, and dual-luciferase reporter gene assay showed that miR-380-3p blocked the translation of the transcription factor Sp3 in the absence of degradation of Sp3 mRNA. Results based on cell counting analysis, annexin v-fluorescein isothiocyanate/propidium iodide doublestaining assay and propidium iodide staining showed that overexpression of miR-380-3p inhibited cell proliferation, increased the apoptotic rate, induced cell cycle arrest and intensified the toxicity of PQ in mouse neuroblastoma（N2 a） cells. Knockdown of Sp3 inhibited cell proliferation and eclipsed the alterations induced by miR-380-3p in cell proliferation. Two mediators of apoptosis and cell cycle identified in previous studies as Sp3-regulated, namely cyclin-dependent kinase inhibitor 1（p21） and calmodulin（CaM）, were dysregulated by PQ, but not Sp3 deficiency. In conclusion, Nrf2-regulated miR-380-3p inhibited cell proliferation and enhanced the PQ-induced toxicity in N2a cells potentially by blocking the translation Sp3 mRNA. We conclude that CaM and p21 were involved in PQ-induced toxicity.