HBx通过抑制microRNA-145a-5p促进小鼠肝癌发生

杜彬; 周梦瑶; 毋楠; 张思遥; 黄欣; 况钦; 吴勇; 冯涛

doi:10.16016/j.1000-5404.202102040

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HBx通过抑制microRNA-145a-5p促进小鼠肝癌发生

基础医学 | 更新时间：2026-01-09

- HBx通过抑制microRNA-145a-5p促进小鼠肝癌发生
- HBx promotes liver tumor in mice by inhibiting microRNA-145a-5p
- 第三军医大学学报 2021年43卷第18期页码：1751-1761
- 作者机构：
  
  400016 重庆，重庆医科大学：分子医学与肿瘤研究中心，药学院生物化学与分子生物研究中心
- 作者简介：
  
  [冯涛，E-mail: 100733@cqmu.edu.cn FENG Tao, E-mail: 100733@cqmu.edu.cn
- 基金信息：
- DOI：10.16016/j.1000-5404.202102040
  中图分类号： R394.3;R730.23;R735.7
- 收稿：2021-02-05，
  
  修回：2021-04-30，
  
  纸质出版：2021-09-30
- 稿件说明：
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杜彬, 周梦瑶, 毋楠, 等. HBx通过抑制microRNA-145a-5p促进小鼠肝癌发生[J]. 第三军医大学学报, 2021,43(18):1751-1761.

Bin DU, Mengyao ZHOU, Nan WU, et al. HBx promotes liver tumor in mice by inhibiting microRNA-145a-5p[J]. Journal of Third Military Medical University, 2021, 43(18): 1751-1761.
杜彬, 周梦瑶, 毋楠, 等. HBx通过抑制microRNA-145a-5p促进小鼠肝癌发生[J]. 第三军医大学学报, 2021,43(18):1751-1761. DOI： 10.16016/j.1000-5404.202102040.

Bin DU, Mengyao ZHOU, Nan WU, et al. HBx promotes liver tumor in mice by inhibiting microRNA-145a-5p[J]. Journal of Third Military Medical University, 2021, 43(18): 1751-1761. DOI： 10.16016/j.1000-5404.202102040.

摘要

目的

通过研究miRNA-145a-5p在肝前体细胞和小鼠感染乙肝病毒X(hepatitis B virus X，HBx)不同时间的变化情况，初步探讨HBx导致肝癌发生的机制。

方法

通过慢病毒将HBx基因转染到14-19肝前体细胞中，构建HBx-EGFP-14-19细胞系，并通过肝门静脉将HBx-EGFP-14-19细胞悬液注入KM小鼠体内，建立具有完整免疫系统的HBx小鼠模型，并分别于30、90、180、360 d收集肝脏组织。通过qRT-PCR和Western blot检测HBx在HBx-EGFP-14-19细胞和HBx小鼠中的表达。将miR-145a-5p mimics和inhibitor转染到HBx-EGFP-14-19细胞和饲养180 d HBx小鼠中，qRT-PCR检测miR-145a-5p、c-Myc、Kras、细胞凋亡和细胞周期因子的RNA表达，Western blot检测c-Myc、Kras、细胞凋亡和细胞周期因子的蛋白质表达，CCK-8检测细胞增殖。

结果

成功构建能够稳定表达HBx的细胞模型和动物模型。HBx过表达后，miR-145a-5p在HBx-EGFP-14-19细胞和HBx小鼠中的表达下调，而c-Myc和Kras的表达升高。使用miR-145a-5p模拟物(mimic/agomir)可以抑制细胞增殖并下调c-Myc、Kras、Bcl-2、CDK4、Cyclin E、CyclinD1的表达，并上调Bad和Bax的表达，而miR-145a-5p抑制剂(inhibitor/antagomir)与miR-145a-5p模拟物组的结果相反。

结论

HBx过表达导致小鼠肝癌发生，其机制可能与HBx调节miR-145a-5p及其靶基因c-Myc和Kras的表达和抑制细胞凋亡有关。

Abstract

Objective

To investigate the changes of miRNA-145a-5p in liver progenitor cells and mice infected with hepatitis B virus X protein (HBx) at different time points to preliminarily explore the mechanism of HBx causing liver tumor.

Methods

HBx gene was transfected into 14-19 liver precursor cells by lentivirus vector to construct HBx-EGFP-14-19 cell line

and then the suspension of obtained cells was injected into KM mice through hepatic portal vein to establish an HBx model with a complete immune system. The liver tissues were collected on days 30

180

and 360. The expression of HBx in HBx-EGFP-14-19 cells and HBx mice was detected by qRT-PCR and Western blotting. After miR-145a-5p mimic and inhibitor was transfected into the HBx-EGFP-14-19 cells and HBx mice having been fed for 180 d

respectively

the RNA expression of miR-145a-5p

c-Myc

Kras

apoptosis-related and cell cycle-related factors was detected by qRT-PCR

and the protein expression of them were detected by Western blotting. The cell proliferation was detected by CCK8 assay.

Results

The cell model capable of stably expressing HBx and HBx mouse model were established successfully. Overexpression of HBx induced the down-regulation of miR-145a-5p in HBx-EGFP-14-19 cells and HBx mice

while enhanced the expression of c-Myc and Kras. MiR-145a-5p mimic/agomir inhibited cell proliferation and down-regulated the expression of c-Myc

Kras

Bcl-2

CDK4

CyclinE

CyclinD1

and up-regulated the expression of Bad and Bax

while miR-145a-5p inhibitor/antagomir led to the opposite Results from the mimic.

Conclusion

Overexpression of HBx leads to the occurrence of liver tumor in mice

and its mechanism may be related to its regulation of miR-145a-5p and its target genes c-Myc and Kras and inhibition for cell apoptosis.

关键词

Keywords

references

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