[1]戴旭芳,秦利燕,连继勤.丙戊酸钠下调SH-SY5Y细胞中Bcl-2表达的作用及机制分析[J].第三军医大学学报,2018,40(12):1060-1066.
 DAI Xufang,QIN Liyan,LIAN Jiqin.Sodium valprovate upregulates miR-34a to inhibit Bcl-2 expression in human neuroblastoma SHSY5Y cells in vitro[J].J Third Mil Med Univ,2018,40(12):1060-1066.
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丙戊酸钠下调SH-SY5Y细胞中Bcl-2表达的作用及机制分析(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第12期
页码:
1060-1066
栏目:
基础医学
出版日期:
2018-06-30

文章信息/Info

Title:
Sodium valprovate upregulates miR-34a to inhibit Bcl-2 expression in human neuroblastoma SHSY5Y cells in vitro
作者:
戴旭芳秦利燕连继勤
重庆师范大学教育科学学院:重庆市特殊儿童心理诊断与教育技术重点实验室,特殊教育系;陆军军医大学(第三军医大学):第一附属医院输血科,基础医学院生物化学与分子生物学教研室
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
 

关键词:
丙戊酸钠SH-SY5Y细胞Bcl-2miR-34a
Keywords:
sodium valproate SH-SY5Y cells Bcl-2 miR-34a
分类号:
R394.6;R749.94;R971.6
文献标志码:
A
摘要:

目的探讨丙戊酸钠(sodium valproate,VPA)在神经细胞中下调Bcl-2表达的作用及可能机制。方法用不同浓度的VPA处理SHSY5Y细胞,定量PCR检测Bcl-2 mRNA与miR34a水平变化,Western blot检测Bcl-2蛋白水平变化,报告基因系统检测Bcl-2启动子活性变化,联合转录抑制剂处理细胞,分析Bcl-2 mRNA稳定性变化,联合miR-34a模拟物及抑制剂处理细胞,进一步分析miR-34a在VPA下调Bcl-2表达中的作用。Annexin V/PI双染结合流式分析细胞凋亡情况。结果VPA可浓度依赖性下调SHSY5Y细胞中Bcl-2的 mRNA与蛋白水平(P<0.05)。VPA暴露不改变Bcl-2的启动子活性(P>0.05),但显著降低了其mRNA的稳定性(P<0.05)。同时,VPA可上调SHSY5Y细胞中miR-34a的表达(P<0.05),联合使用miR-34a的抑制剂可逆转VPA对Bcl2表达的下调作用,且单独的miR-34a模拟物也可以下调细胞中Bcl2的表达。VPA暴露还可引起SH-SY5Y细胞的凋亡增加(P<0.05)。结论〓〖HTSS〗VPA可通过上调SH-SY5Y细胞中miR-34a水平而抑制Bcl2的表达。

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. 
 

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更新日期/Last Update: 2018-07-02