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



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


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


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


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. 


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Last Update: 2018-07-02