|Table of Contents|

Sodium valprovate upregulates miR-34a to inhibit Bcl-2 expression in human neuroblastoma SHSY5Y cells in vitro

(PDF)

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

Issue:
2018年第12期
Page:
1060-1066
Research Field:
基础医学
Publishing date:

Info

Title:

Sodium valprovate upregulates miR-34a to inhibit Bcl-2 expression in human neuroblastoma SHSY5Y cells in vitro

Author(s):

DAI Xufang QIN LiyanLIAN Jiqin

Chongqing Key Laboratory of Psychological Diagnosis and Educational Technology for Children with Special Needs, Faculty of Education for Children with Special Needs, College of Education Science, Chongqing Normal University, Chongqing, 400047; Department of Blood Transfusion, First Affiliated Hospital,Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing, 400038, China
 

Keywords:

sodium valproate SH-SY5Y cells Bcl-2 miR-34a

PACS:
R394.6;R749.94;R971.6
DOI:
-
Abstract:

AbstractObjectiveTo investigate the inhibitory effect of sodium valproate (VPA) exposure on Bcl-2 expression and explore the possible mechanism in human neuroblastoma SH-SY5Y cells in vitro. Methods SH-0SY5Y cells exposed to different concentrations of VPA were examined for expression levels of bcl-2 mRNA and miR34a using qRT-PCR and for Bcl-2 protein expression using Western blotting. The changes in the promoter activity of bcl-2 gene in VPAtreated cells were analyzed using a reporter gene assay, and the stability of bcl-2 mRNA was evaluated after transcription inhibition. A miR34a mimic and a miR-34a antagonist were used to analyze the role of miR-34a in VPAinduced Bcl-2 downregulation in the cells. The apoptosis of SHSY5Y cells following VPA exposures was assessed using flow cytometry with Annexin V and PI staining.Results exposure dosedependently downregulated the expression of Bcl-2 at both the mRNA and protein levels in SHSY5Y cells (P<0.05). VPA treatment significantly decreased the stability of bcl-2 mRNA (P<0.05) without affecting its transcriptional activity (P>0.05), and obviously up-regulated the expression of miR-34a in SHSY5Y cells (P<0.05). Treatment of the cells with the miR-34a mimic alone inhibited the expression of Bcl-2, and application of the miR-34a antagonist obviously reversed the inhibitory effect of VPA on Bcl-2 expression in SH-SY5Y cells. VPA exposure also significantly increased the apoptosis of SH-SY5Y cells (P<0.05). Conclusion VPA exposure down-regulates the expression of Bcl-2 possibly by up-regulating miR-34a level in SH-SY5Y cells. 
 

References:

[1]CAREAGA M, MURAI T, BAUMAN M D. Maternal immune activation and autism spectrum disorder: from rodents to nonhuman and human primates[J]. Biol Psychiatry, 2017, 81(5): 391-401. DOI: 10.1016/j.biopsych.2016.10.020. 
[2]CHOMIAK T, TURNER N, HU B. What we have learned about autismspectrum disorder from valproic acid[J]. Patholog Res Int, 2013, 2013: 712758. DOI: 10.1155/ 2013/712758.
[3]ROULLET F I, LAI J K, FOSTER J A. In utero exposure to valproic acid and autism—a current review of clinical and animal studies[J]. Neurotoxicol Teratol, 2013, 36: 47-56. DOI: 10.1016/j.ntt.2013.01.004.
[4]ZHENG J H, VIACAVA FOLLIS A, KRIWACKI R W, et al. Discoveries and controversies in BCL2 proteinmediated apoptosis[J]. FEBS J, 2016, 283(14): 2690-2700. DOI: 10.1111/febs.13527. 
[5]MALIK M, TAUQEER Z, SHEIKH A M, et al. NFκB signaling in the brain of autistic subjects[J]. Mediators Inflamm, 2011, 2011: 785265. DOI: 10.1155/2011/785265.
[6]Crider A,Ahmed A O,Pillai A,et al.Altered expression of endoplasmic reticulum stressrelated genes in the middle frontal cortex of subjects with autism spectrum disorder[J]. Mol Neuropsychiatr, 2017, 3(2): 85-91. DOI: 10.1159/ 000477212.
[7]CHO H, KIM C H, KNIGHT E Q, et al. Changes in brain metabolic connectivity underlie autisticlike social deficits in a rat model of autism spectrum disorder[J]. Sci Rep, 2017, 7(1): 13213. DOI:  10.1038/s41598017136423.
[8]GAO J, WANG X, SUN H, et al. Neuroprotective effects of docosahexaenoic acid on hippocampal cell death and learning and memory impairments in a valproic acidinduced rat autism model[J]. Int J Dev Neurosci, 2016, 49: 67-78. DOI: 10.1016/j.ijdevneu.2015.11.006. 
[9]HU G W ,MCQUISTON T,BERNARD A, et al. A conserved mechanism of TORdependent RCKmediated mRNA degradation regulates autophagy[J]. Nat Cell Biol, 2015, 17(7): 930-942. DOI: 10.1038/ncb3189.〖ZK)〗
[10]YANG E J, AHN S, LEE K, et al. Correction: early behavioral abnormalities and perinatal alterations of PTEN/AKT pathway in valproic acid autism model mice[J]. PLoS ONE, 2016, 11(6): e0157202. DOI:10.1371/journal. pone.0157202
[11]TONG J, FURUKAWA Y, SHERWIN A, et al. Heterogeneous intrastriatal pattern of proteins regulating axon growth in normal adult human brain[J]. Neurobiol Dis, 2011, 41(2): 458-468. DOI: 10.1016/j.nbd.2010.10.017.
[12]COX E T, BRENNAMAN L H, GABLE K L, et al. Developmental regulation of neural cell adhesion molecule in human prefrontal cortex[J]. Neuroscience, 2009, 162(1): 96-105. DOI: 10.1016/j.neuroscience.2009.04.037. 
[13]LIU M E, HUANG C C, HWANG J P, et al. Effect of Bcl2 rs956572 SNP on regional gray matter volumes and cognitive function in elderly males without dementia[J]. Age (Dordr), 2013, 35(2): 343-352. DOI: 10.1007/s11357-01193675. 
[14]PARK H A, LICZNERSKI P, ALAVIAN K N, et al. BclxL is necessary for neurite outgrowth in hippocampal neurons[J]. Antioxid Redox Signal, 2015, 22(2): 93-108. DOI: 10.1089/ ars.2013.5570. 
[15]YUN Y C,JANG D,YOON S B,et al. Laser acupuncture exerts neuroprotective effects via regulation of creb, bdnf, Bcl2, and bax gene expressions in the hippocampus [J]. Evid Based Complement Alternat Med, 2017, 2017: 7181637. DOI: 10.1155/2017/7181637.
[16]TAKAHASHI M, ISHIDA M, SAITO T, et al. Valproic acid downregulates Cdk5 activity via the transcription of the p35 mRNA[J]. Biochem Biophys Res Commun, 2014, 447(4): 678-682. DOI: 10.1016/j.bbrc.2014.04.072. 
[17]JONAS S, IZAURRALDE E. Towards a molecular understanding of microRNAmediated gene silencing[J]. Nat Rev Genet, 2015, 16(7): 421-433. DOI: 10.1038/nrg3965.

Memo

Memo:
-
Last Update: 2018-07-02