Abstract
Long non-coding RNAs (lncRNAs) are bound up with various human diseases. However, their roles in brain ischemia-reperfusion (I/R) injury remain largely unknown. This study aimed to reveal the potential mechanism of LncRNA SNHG3 on autophagy-induced neuronal cell apoptosis in the brain I/R injury. LncRNA SNHG3 and miR-485 or autophagy markers LC3II/I and Beclin-1 expressions were detected by qRT-PCR or Western blot and the apoptosis of N2a cells was analyzed by flow cytometry. Besides, the interactions between LncRNA SNHG3 and miR-485, miR-485 and ATG7 were validated by RNA pull-down and dual-luciferase reporter system assays. After the Oxygen and Glucose Deprivation (OGD) treatment of N2a cells transfected with pcDNA-SNHG3, pcDNA-SNHG3 + miR-485 mimic for 6 h, 1 mM autophagy inhibitor 3-MA was added and reoxygenated for 24 h, the effect of LncRNA SNHG3 on the autophagy-induced neuronal cell apoptosis was measured by Western blot and flow cytometry. LncRNA SNHG3 was highly expressed in the mouse model of transient middle cerebral artery occlusion and cell model of Oxygen and Glucose Deprivation/Reperfusion, while miR-485 was lowly expressed. Furthermore, miR-485 negatively regulated the luciferase activities of LncRNA SNHG3 and ATG7. After the OGD treatment of N2a cells transfected with pcDNA-SNHG3, pcDNA-SNHG3 + miR-485 mimic for 6 h, 1 mM 3-MA was added and reoxygenated for 24 h, the overexpression of LncRNA SNHG3 raised the ratio of LC3-II/LC3-I and Beclin-1 expression and boosted the apoptosis of N2a cells, while these effects were reversed after the transfection of miR-485 mimic. In general, our data expounded that the interference with LncRNA SNHG3 improved brain I/R injury by up-regulating miR-485 and down-regulating ATG7 to restrain autophagy and neuronal cell apoptosis.
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References
Achawanantakun R, Chen J, Sun Y, Zhang Y (2015) LncRNA-ID: long non-coding RNA IDentification using balanced random forests. Bioinformatics (Oxford, England) 31:3897–3905. https://doi.org/10.1093/bioinformatics/btv480
Batista PJ, Chang HY (2013) Long noncoding RNAs: cellular address codes in development and disease. Cell 152:1298–1307. https://doi.org/10.1016/j.cell.2013.02.012
Cai J et al (2019) Knockdown of lncRNA Gm11974 protect against cerebral ischemic reperfusion through miR-766-3p/NR3C2 axis. Artificial cells, nanomedicine, and biotechnology 47:3847–3853. https://doi.org/10.1080/21691401.2019.1666859
Cesana M et al (2011) A long noncoding RNA controls muscle differentiation by functioning as a competing endogenous RNA. Cell 147:358–369. https://doi.org/10.1016/j.cell.2011.09.028
Chen F, Zhang L, Wang E, Zhang C, Li X (2018) LncRNA GAS5 regulates ischemic stroke as a competing endogenous RNA for miR-137 to regulate the Notch1 signaling pathway. Biochemical and biophysical research communications 496:184–190. https://doi.org/10.1016/j.bbrc.2018.01.022
Duehrkop C, Rieben R (2014) Ischemia/reperfusion injury: effect of simultaneous inhibition of plasma cascade systems versus specific complement inhibition. Biochemical pharmacology 88:12–22. https://doi.org/10.1016/j.bcp.2013.12.013
Fan CN, Ma L, Liu N (2018) Systematic analysis of lncRNA-miRNA-mRNA competing endogenous RNA network identifies four-lncRNA signature as a prognostic biomarker for breast cancer. Journal of translational medicine 16:264. https://doi.org/10.1186/s12967-018-1640-2
Flintoft L (2013) Non-coding RNA: Structure and function for lncRNAs. Nat Rev Genet 14:598. https://doi.org/10.1038/nrg3561
Fok ET, Scholefield J, Fanucchi S, Mhlanga MM (2017) The emerging molecular biology toolbox for the study of long noncoding RNA biology. Epigenomics 9:1317–1327. https://doi.org/10.2217/epi-2017-0062
He G, Xu W, Tong L, Li S, Su S, Tan X, Li C (2016) Gadd45b prevents autophagy and apoptosis against rat cerebral neuron oxygen-glucose deprivation/reperfusion injury. Apoptosis : an international journal on programmed cell death 21:390–403. https://doi.org/10.1007/s10495-016-1213-x
Kondo Y, Shinjo K, Katsushima K (2017) Long non-coding RNAs as an epigenetic regulator in human cancers. Cancer science 108:1927–1933. https://doi.org/10.1111/cas.13342
Li S et al (2017) Complex integrated analysis of lncRNAs-miRNAs-mRNAs in Oral squamous cell carcinoma. Oral oncology 73:1–9. https://doi.org/10.1016/j.oraloncology.2017.07.026
Liang H et al (2019) LncRNA 2810403D21Rik/Mirf promotes ischemic myocardial injury by regulating autophagy through targeting Mir26a. Autophagy:1–15. https://doi.org/10.1080/15548627.2019.1659610
Mai C, Qiu L, Zeng Y, Jian HG (2019) LncRNA Lethe protects sepsis-induced brain injury via regulating autophagy of cortical neurons. European review for medical and pharmacological sciences 23:4858–4864. https://doi.org/10.26355/eurrev_201906_18073
Park C, Yu N, Choi I, Kim W, Lee S (2014) lncRNAtor: a comprehensive resource for functional investigation of long non-coding RNAs. Bioinformatics (Oxford, England) 30:2480–2485. https://doi.org/10.1093/bioinformatics/btu325
Qiang L, Sample A, Shea CR, Soltani K, Macleod KF, He YY (2017) Autophagy gene ATG7 regulates ultraviolet radiation-induced inflammation and skin tumorigenesis. Autophagy 13:2086–2103. https://doi.org/10.1080/15548627.2017.1380757
Qin AP et al (2010) Autophagy was activated in injured astrocytes and mildly decreased cell survival following glucose and oxygen deprivation and focal cerebral ischemia. Autophagy 6:738–753. https://doi.org/10.4161/auto.6.6.12573
Qiu-li Z, Wei W (2018) LncRNA-H19 Induces Retinal Müller Cell Apoptosis via MiR-29b/FOXO4 Axis in Diabetic Retinopathy Clinical surgery research communications 2 doi:https://doi.org/10.31491/csrc.2018.12.024
Sukseree S et al (2018) Filamentous aggregation of Sequestosome-1/p62 in brain neurons and Neuroepithelial cells upon Tyr-Cre-mediated deletion of the autophagy gene Atg7. Molecular neurobiology 55:8425–8437. https://doi.org/10.1007/s12035-018-0996-x
Tay Y, Rinn J, Pandolfi PP (2014) The multilayered complexity of ceRNA crosstalk and competition. Nature 505:344–352. https://doi.org/10.1038/nature12986
Turner RC, Dodson SC, Rosen CL, Huber JD (2013) The science of cerebral ischemia and the quest for neuroprotection: navigating past failure to future success. Journal of neurosurgery 118:1072–1085. https://doi.org/10.3171/2012.11.Jns12408
Wang HJ et al (2018a) Endothelial Atg7 deficiency ameliorates acute cerebral injury induced by ischemia/reperfusion. Frontiers in neurology 9:998. https://doi.org/10.3389/fneur.2018.00998
Wang J, Cao B, Han D, Sun M, Feng J (2017) Long non-coding RNA H19 induces cerebral ischemia reperfusion injury via activation of autophagy. Aging and disease 8:71–84. https://doi.org/10.14336/ad.2016.0530
Wang M et al (2018b) Long non-coding RNA H19 confers 5-Fu resistance in colorectal cancer by promoting SIRT1-mediated autophagy. Cell death & disease 9:1149. https://doi.org/10.1038/s41419-018-1187-4
Wang SH et al (2016) Long non-coding RNA H19 regulates FOXM1 expression by competitively binding endogenous miR-342-3p in gallbladder cancer. Journal of experimental & clinical cancer research: CR 35:160. https://doi.org/10.1186/s13046-016-0436-6
Wei R, Zhang L, Hu W, Wu J, Zhang W (2019) Long non-coding RNA AK038897 aggravates cerebral ischemia/reperfusion injury via acting as a ceRNA for miR-26a-5p to target DAPK1. Experimental neurology 314:100–110. https://doi.org/10.1016/j.expneurol.2019.01.009
Wen XR et al (2016) Butylphthalide suppresses neuronal cells apoptosis and inhibits JNK-Caspase3 signaling pathway after brain ischemia /reperfusion in rats. Cellular and molecular neurobiology 36:1087–1095. https://doi.org/10.1007/s10571-015-0302-7
Wu Z et al (2017) LncRNA-N1LR enhances Neuroprotection against ischemic stroke probably by inhibiting p53 phosphorylation. Molecular neurobiology 54:7670–7685. https://doi.org/10.1007/s12035-016-0246-z
Xie C et al (2016) Neuroprotection by selective neuronal deletion of Atg7 in neonatal brain injury. Autophagy 12:410–423. https://doi.org/10.1080/15548627.2015.1132134
Xiong J (2015) Atg7 in development and disease: panacea or Pandora's box? Protein & cell 6:722–734. https://doi.org/10.1007/s13238-015-0195-8
Xu J et al (2015) The mRNA related ceRNA-ceRNA landscape and significance across 20 major cancer types. Nucleic acids research 43:8169–8182. https://doi.org/10.1093/nar/gkv853
Yan H, Rao J, Yuan J, Gao L, Huang W, Zhao L, Ren J (2017) Long non-coding RNA MEG3 functions as a competing endogenous RNA to regulate ischemic neuronal death by targeting miR-21/PDCD4 signaling pathway. Cell death & disease 8:3211. https://doi.org/10.1038/s41419-017-0047-y
Yan W, Chen ZY, Chen JQ, Chen HM (2018) LncRNA NEAT1 promotes autophagy in MPTP-induced Parkinson's disease through stabilizing PINK1 protein. Biochemical and biophysical research communications 496:1019–1024. https://doi.org/10.1016/j.bbrc.2017.12.149
Yang L, Peng X, Jin H, Liu J (2019) Long non-coding RNA PVT1 promotes autophagy as ceRNA to target ATG3 by sponging microRNA-365 in hepatocellular carcinoma. Gene 697:94–102. https://doi.org/10.1016/j.gene.2019.02.036
Yu S, Yu M, He X, Wen L, Bu Z, Feng J (2019) KCNQ1OT1 promotes autophagy by regulating miR-200a/FOXO3/ATG7 pathway in cerebral ischemic stroke. Aging cell 18:e12940. https://doi.org/10.1111/acel.12940
Zhou Z, Xu H, Liu B, Dun L, Lu C, Cai Y, Wang H (2019) Suppression of lncRNA RMRP ameliorates oxygen-glucose deprivation/re-oxygenation-induced neural cells injury by inhibiting autophagy and PI3K/Akt/mTOR-mediated apoptosis. Bioscience reports:39. https://doi.org/10.1042/bsr20181367
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Cao, Y., Pan, L., Zhang, X. et al. LncRNA SNHG3 promotes autophagy-induced neuronal cell apoptosis by acting as a ceRNA for miR-485 to up-regulate ATG7 expression. Metab Brain Dis 35, 1361–1369 (2020). https://doi.org/10.1007/s11011-020-00607-1
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DOI: https://doi.org/10.1007/s11011-020-00607-1