Sodium valprovate upregulates miR-34a to inhibit Bcl-2 expression in human neuroblastoma SH-SY5Y cells <italic style="font-style: italic">in vitro</italic>

Xufang DAI; Liyan QIN; Jiqin LIAN

doi:10.16016/j.1000-5404.201712088

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

Basic Medicine | 更新时间：2026-01-09

- Sodium valprovate upregulates miR-34a to inhibit Bcl-2 expression in human neuroblastoma SH-SY5Y cells in vitro
- Journal of Third Military Medical University Vol. 40, Issue 12, Pages: 1060-1065(2018)
- 作者机构：
  
  1.400047 重庆，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
  2.00047 重庆，重庆师范大学教育科学学院:特殊教育系
  3.400038 重庆，陆军军医大学(第三军医大学)：第一附属医院输血科
  4.400038 重庆，陆军军医大学(第三军医大学)：基础医学院生物化学与分子生物学教研室
- 作者简介：
- 基金信息：
  
  the National Natural Science Foundation for Young Scholars of China(31201068)
- DOI：10.16016/j.1000-5404.201712088
  CLC： R394.6; R749.94; R971.6
- Received：11 December 2017，
  
  Revised：2018-02-12，
  
  Published：2018-01
- 稿件说明：
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Xufang DAI, Liyan QIN, Jiqin LIAN. Sodium valprovate upregulates miR-34a to inhibit Bcl-2 expression in human neuroblastoma SH-SY5Y cells in vitro[J]. Journal of Third Military Medical University, 2018, 40(12): 1060-1065.
DOI：

Xufang DAI, Liyan QIN, Jiqin LIAN. Sodium valprovate upregulates miR-34a to inhibit Bcl-2 expression in human neuroblastoma SH-SY5Y cells in vitro[J]. Journal of Third Military Medical University, 2018, 40(12): 1060-1065. DOI： 10.16016/j.1000-5404.201712088.

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(

0.05)。VPA暴露不改变Bcl-2的启动子活性(

0.05)，但显著降低了其mRNA的稳定性(

0.05)。同时，VPA可上调SH-SY5Y细胞中miR-34a的表达(

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(

0.05)。

结论

VPA可通过上调SH-SY5Y细胞中miR-34a水平而抑制Bcl-2的表达。

Abstract

Objective

To 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-SY5Y 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

VPA exposure dose-dependently down-regulated the expression of Bcl-2 at both the mRNA and protein levels in SH-SY5Y cells (

0.05). VPA treatment significantly decreased the stability of bcl-2 mRNA (

0.05) without affecting its transcriptional activity (

0.05)

and obviously up-regulated the expression of miR-34a in SH-SY5Y cells (

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 (

0.05).

Conclusion

VPA exposure down-regulates the expression of Bcl-2 possibly by up-regulating miR-34a level in SH-SY5Y cells.

关键词

Keywords

references

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

DOI：10.16016/j.1000-5404.201712088

摘要

Abstract

关键词

Keywords

references