HBx promotes liver tumor in mice by inhibiting microRNA-145a-5p

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HBx promotes liver tumor in mice by inhibiting microRNA-145a-5p

Basic Medicine | 更新时间：2026-01-09

- HBx promotes liver tumor in mice by inhibiting microRNA-145a-5p
- Journal of Third Military Medical University Vol. 43, Issue 18, Pages: 1751-1761(2021)
- 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：
- 基金信息：
- DOI：10.16016/j.1000-5404.202102040
  CLC： R394.3;R730.23;R735.7
- Received：05 February 2021，
  
  Revised：2021-04-30，
  
  Published：30 September 2021
- 稿件说明：
移动端阅览
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：

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

2

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

2

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

2

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

2

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

2

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

2

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

90

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

2

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

2

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

BRAY F, FERLAY J, SOERJOMATARAM I, et al . Global cancer statistic s 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J ] . CA Cancer J Clin, 2018, 68(6): 394-424. DOI: 10.3322/caac.21492.

Yongyan Chen, Zhigang Tian. HBV-Induced Immune Imbalance in the Development of HCC[J]. Front Immunol, 2019, 10: 2048. DOI: 10.3389/fimmu.2019.02048.

LEVRERO M, ZUCMAN-ROSSI J. Mechanisms of HBV-induced hepatocellular carcinoma[J]. J Hepatol, 2016, 64(Suppl 1): S84-S101. DOI: 10.1016/j.jhep.2016.02.021.

CHEN W S, YEN C J, CHEN Y J, et al . miRNA-7/21/107 contribute to HBx-induced hepatocellular carcinoma progression through suppression of maspin[J ] . Oncotarget, 2015, 6(28): 25962-25974. DOI: 10.18632/oncotarget.4504.

GAO F, SUN X Y, WANG L K, et al . Downregulation of MicroRNA-145 caused by hepatitis B virus X protein promotes expression of CUL5 and contributes to pathogenesis of hepatitis B virus-associated hepatocellular carcinoma[J ] . Cell Physiol Biochem, 2015, 37(4): 1547-1559. DOI: 10.1159/000438522.

LI J F, DAI X P, ZHANG W, et al . Upregulation of microRNA-146a by hepatitis B virus X protein contributes to he patitis development by downregulating complement factor H[J ] . mBio, 2015, 6(2): e02459-e02414. DOI: 10.1128/mBio.02459-14.

CHEN Z, ZENG H Z, GUO Y, et al . miRNA-145 inhibits non-small cell lung cancer cell proliferation by targeting c-Myc[J ] . J Exp Clin Cancer Res, 2010, 29: 151. DOI: 10.1186/1756-9966-29-151.

YU Y J, NANGIA-MAKKER P, FARHANA L, et al . miR-21 and miR-145 cooperation in regulation of colon cancer stem cells[J ] . Mol Cancer, 2015, 14: 98. DOI: 10.1186/s12943-015-0372-7.

SADEGHIYEH N, SEHATI N, MANSOORI B, et al . MicroRNA-145 replacement effect on growth and migration inhibition in lung cancer cell line[J ] . Biomed Pharmacother, 2019, 111: 460-467. DOI: 10.1016/j.biopha.2018.12.094.

KAZI A, XIANG S Y, YANG H, et al . GSK3 suppression upregulates β-catenin and c-Myc to abrogate KRas-dependent tumors[J ] . Nat Commun, 2018, 9(1): 5154. DOI: 10.1038/s41467-018-07644-6.

BANDOPADHYAY M, BANERJEE A, SARKAR N, et al . Tumor suppressor microRNA miR-145 and onco microRNAs miR-21 and miR-222 expressions are differentially modulated by hepat itis B virus X protein in malignant hepatocytes[J ] . BMC Cancer, 2014, 14: 721. DOI: 10.1186/1471-2407-14-721.

GAI X C, TANG B F, LIU F M, et al . mTOR/miR-145-regulated exosomal GOLM1 promotes hepatocellular carcinoma through augmented GSK-3β/MMPs[J ] . J Genet Genom, 2019, 46(5): 235-245. DOI: 10.1016/j.jgg.2019.03.013.

DU W B, LI D, GUO X M, et al . Circ-PRMT5 promotes gastric cancer progression by sponging miR-145 and miR-1304 to upregulate MYC[J ] . Artif Cells Nanomed Biotechnol, 2019, 47(1): 4120-4130. DOI: 10.1080/21691401.2019.1671857.

WANG X, HUO B N, LIU J, et al . Hepatitis B virus X reduces hepatocyte apoptosis and promotes cell cycle progression through the Akt/mTOR pathway in vivo[J ] . Gene, 2019, 691: 87-95. DOI: 10.1016/j.gene.2018.12.054.

CHATURVEDI V K, SINGH A, DUBEY S K, et al . Molecular mechanistic insight of hepatitis B virus mediated hepatocellular carcinoma[J ] . Microb Pathog, 2019, 128: 184-194. DOI: 10.1016/j.micpath.2019.01.004.

CHEN Y Y, TIAN Z G. HBV-induced immune imbalance in the development of HCC[J]. Front Immunol, 2019, 10: 2048. DOI: 10.3389/fimmu.2019.02048.

ALI A, ABDEL-HAFIZ H, SUHAIL M, et al . Hepatitis B virus, HBx mutants and their role in hepatocellular carcinoma[J ] . World J Gastroenterol, 2014, 20(30): 10238-10248. DOI: 10.3748/wjg.v20.i30.10238.

KIM D W, TALATI C, KIM R. Hepatocellular carcinoma (HCC): beyond sorafenib-chemotherapy[J]. J Gastrointest Oncol, 2017, 8(2): 256-265. DOI: 10.21037/jgo.2016.09.07.

LUO P, WU S Y, YU Y L, et al . Current status and perspective biomarkers in AFP negative HCC: towards screening for and diagnosing hepatocellular carcinoma at an earlier stage[J ] . Pathol Oncol Res, 2020, 26(2): 599-603. DOI: 10.1007/s12253-019-00585-5.

MISHRA S, YADAV T, RANI V. Exploring miRNA based approaches in cancer diagnostics and therapeutics[J]. Crit Rev Oncol Hematol, 2016, 98: 12-23. DOI: 10.1016/j.critrevonc.2015.10.003.

FU Y H, YANG X, CHEN H Y, et al . Diagnostic value of miR-145 and its regulatory role in macrophage immune response in tuberculosis[J ] . Genet Mol Biol, 2020, 43(2): e20190238. DOI: 10.1590/1678-4685-GMB-2019-0238.

HE M, WU N, LEONG M C, et al . miR-145 improves metabolic inflammatory disease through multiple pathways[J ] . J Mol Cell Biol, 2020, 12(2): 152-162. DOI: 10.1093/jmcb/mjz015.

DANG C V. MYC on the path to cancer[J]. Cell, 2012, 149(1): 22-35. DOI: 10.1016/j.cell.2012.03.003.

LIU P, GE M M, HU J J, et al . A functional mammalian target of rapamycin complex 1 signaling is indispensable for c-Myc-driven hepatocarcinogenesis[J ] . Hepatology, 2017, 66(1): 167-181. DOI: 10.1002/hep.29183.

SHACHAF C M, KOPELMAN A M, ARVANITIS C, et al . MYC inactivation uncovers pluripotent differentiation and tumour dormancy in hepatocellular cancer[J ] . Nature, 2004, 431(7012): 1112-1117. DOI: 10.1038/nature03043.

CHOW E K, FAN L L, CHEN X, et al . Oncogene-specific formation of chemoresistant murine hepatic cancer stem cells[J ] . Hepatology, 2012, 56(4): 1331-1341. DOI: 10.1002/hep.25776.

KARNOUB A E, WEINBERG R A. Ras oncogenes: split personalities[J]. Nat Rev Mol Cell Biol, 2008, 9(7): 517-531. DOI: 10.1038/nrm2438.

YE H, ZHANG C, WANG B J, et al . Synergistic function of Kras mutation and HBx in initiation and progression of hepatocellular carcinoma in mice[J ] . Oncogene, 2014, 33(43): 5133-5138. DOI: 10.1038/onc.2013.468.

LEDFORD H. Gene-silencing technology gets first drug approval after 20-year wait[J]. Nature, 2018, 560(7718): 291-292. DOI: 10.1038/d41586-018-05867-7.

MULLIGAN M J, LYKE K E, KITCHIN N, et al . Phase Ⅰ/Ⅱ study of COVID-19 RNA vaccine BNT162b1 in adults[J ] . Nature, 2020, 586(7830): 589-593. DOI: 10.1038/s41586-020-2639-4.

WALSH E E, FRENCK R W Jr, FALSEY A R, et al . Safety and immunogenicity of two RNA-based covid-19 vaccine candidates[J ] . N Engl J Med, 2020, 383(25): 2439-2450. DOI: 10.1056/NEJMoa2027906.

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