miRNA-146a对乙型肝炎病毒生活周期的影响及机制研究

赵敏; 张祯祯; 刘泉波

doi:10.16016/j.1000-5404.201702123

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miRNA-146a对乙型肝炎病毒生活周期的影响及机制研究

基础医学 | 更新时间：2026-01-09

- miRNA-146a对乙型肝炎病毒生活周期的影响及机制研究
- Effect of miRNA-146a on life cycle of hepatitis B virus in vitro
- 第三军医大学学报 2017年39卷第17期页码：1702-1708
- 作者机构：
  
  400014 重庆，重庆医科大学附属儿童医院感染科，儿童发育疾病研究教育部重点实验室，儿童发育重大疾病国家国际科技合作基地，儿科学重庆市重点实验室
- 作者简介：
  
  [ 刘泉波，E-mail：liuqb1223@sina.com LIU Quanbo, E-mail: liuqb1223@sina.com
- 基金信息：
  
  国家自然科学基金青年科学基金(81501736);高等学校博士学科点专项科研基金(20135503120008)
- DOI：10.16016/j.1000-5404.201702123
  中图分类号： R373.21;R394-33;R394.3
- 收稿：2017-02-23，
  
  修回：2017-04-25，
  
  纸质出版：2017-09-15
- 稿件说明：
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赵敏, 张祯祯, 刘泉波. miRNA-146a对乙型肝炎病毒生活周期的影响及机制研究[J]. 第三军医大学学报, 2017,39(17):1702-1708.

Min ZHAO, Zhenzhen ZHANG, Quanbo LIU. Effect of miRNA-146a on life cycle of hepatitis B virus in vitro[J]. Journal of Third Military Medical University, 2017, 39(17): 1702-1708.
赵敏, 张祯祯, 刘泉波. miRNA-146a对乙型肝炎病毒生活周期的影响及机制研究[J]. 第三军医大学学报, 2017,39(17):1702-1708. DOI： 10.16016/j.1000-5404.201702123.

Min ZHAO, Zhenzhen ZHANG, Quanbo LIU. Effect of miRNA-146a on life cycle of hepatitis B virus in vitro[J]. Journal of Third Military Medical University, 2017, 39(17): 1702-1708. DOI： 10.16016/j.1000-5404.201702123.

摘要

目的

研究微小RNA-146a(microRNA-146a，miR-146a)对乙型肝炎病毒(hepatitis B virus，HBV)生活周期的影响及其可能机制。

方法

利用miRNA芯片比较HepG2.2.15细胞与HepG2细胞miRNAs表达谱差异，选择miR-146a为研究对象，RT-PCR验证芯片结果。在HepG2.2.15细胞中分别转染miR-146a mimic和inhibitor，RT-PCR检测HBV复制水平

ELISA和Western blot检测蛋白水平。双荧光素酶报告系统进一步验证miR-146a与潜在靶点HS3ST3B1相互作用。HepG2.2.15细胞中转染miR-146a mimic，RT-PCR和Western blot分别检测HS3ST3B1 mRNA和蛋白水平。

结果

miRNA芯片发现72条miRNAs在HepG2.2.15中表达发生变化，其中27条上调，45条下调

RT-PCR证实，miR-146a在HepG2.2.15细胞(1.55±0.13) 中表达水平明显高于HepG2细胞(1.00±0.01) (

0.05)。HepG2.2.15细胞转染miR-146a mimic后，HBV复制和蛋白水平较对照组明显升高(

0.05)；转染miR-146a inhibitor后，HBV复制和蛋白表达水平较对照组明显降低(

0.05)。生物信息学预测发现HBV抑制因子HS3ST3B1是miR-146a的潜在靶点，双荧光素酶报告系统显示

HS3ST3B1野生型载体的报告荧光较对照组明显下调(

0.05)，突变预测靶位点后，HS3ST3B1突变型载体的报告荧光与对照组差异无统计学意义(

0.05)。HepG2.2.15细胞中转染miR-146a mimic后，HS3ST3B1 mRNA水平较对照组差异无统计学意义(

0.05)，HS3ST3B1蛋白水平较对照组明显下调。

结论

miR-146a能影响HBV生活周期，miR-146a可能通过作用于HBV抑制因子HS3ST3B1 3′UTR抑制其翻译从而影响HBV生活周期。

Abstract

Objective

To determine the effect of microRNA-146a (miR-146a) on the life cycle of hepatitis B virus (HBV) and investigate the underlying mechanisms.

Methods

The miRNA expression profiles were compared by miRNA array between HepG2 and HepG2.2.15 cells. Then miR-146a was chosen as objective

and its expression level was further confirmed by RT-PCR. After miR-146a mimic and inhibitor were transfected into HepG2.2.15 cells respectively

the quantification of HBV replication was determined by RT-PCR

and the levels of HBsAg and HBeAg in the supernatant were measured by ELISA

and the expression of HS3ST3B1 at mRNA and protein levels were tested by RT-PCR and Western blotting. Dual-luciferase reporter assay was used to detect the interaction between miR-146a and potential target HS3ST3B1.

Results

The expression levels of totally 72 miRNAs were changed in HepG2.2.15 cells

with 27 up-regulated and 45 down-regulated. RT-PCR showed the expression level of miR-146a was significantly higher in HepG2.2.15 cells than HepG2 cells (1.55±0.13

1.00±0.01

0.05). Transfection of miR-146a mimic into HepG2.2.15 cells resulted in significantly increased HBV replication and levels of HBsAg and HBeAg (

0.05)

while the transfection of its inhibited caused opposite results (

0.05).Bioinformatic analysis showed that HS3ST3B1 was a potential target of miR-146a. The reporter luciferase reporter system indicated that the reported fluorescence intensity of HS3ST3B1 wild type vector was significantly lower than that of the control group (

0.05)

but showed no significant difference between HS3ST3B1 mutant vector and control group (

0.05). The mRNA level of HS3ST3B1 was not significantly changed in HepG2.2.15 cells transfected with miR-146a mimic (

0.05)

but its protein level was significantly decreased (

0.05).

Conclusions

miR-146a affects the life cycle of HBV

which may be through suppressing the translation of HBV inhibitory factor HS3ST3B1 3′UTR.

关键词

Keywords

references

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