Hepatology International

, Volume 13, Issue 6, pp 814–825 | Cite as

MicroRNA-744/transforming growth factor β1 relationship regulates liver cirrhosis

  • Shuang Ren
  • Jiamei Chen
  • Qinglan Wang
  • Xuewei Li
  • Ying Xu
  • Xiao Zhang
  • Yongping Mu
  • Hua Zhang
  • Shuang HuangEmail author
  • Ping LiuEmail author
Original Article



MicroRNAs have added a new dimension to our understanding of liver cirrhosis (LC) and associated processes like the activation of hepatic stellate cells (HSCs).


Serum samples were collected from 40 LC patients and 30 healthy donors. CCl4-induced LC mouse model in vivo and in vitro human HSC LX-2 and murine HSC JS-1 cells were researched.


The levels of serum microRNA (miR)-744 is inversely correlated with the severity of LC and is a reliable biomarker of LC. In CCl4-induced LC model, the abundance of miR-744 was reduced in both sera and livers compared with sham controls. Importantly, increasing miR-744 abundance with synthetic miR-744 Agomir alleviated liver fibrosis, a critical component of LC, while reducing miR-744 with Antagomir exacerbated it. To elucidate molecular mechanism underlying the suppressive role of miR-744 in LC, we observed that miR-744 and transforming growth factor β1 (TGFβ1) are inversely correlated in LC patients’ sera as well as sera/livers from CCl4-induced LC mice. We demonstrated that miR-744 Agomir downregulated the expression of TGFβ1 and further confirmed that TGFβ1 mRNA was a bona fide miR-744 target in HSCs. Moreover, miR-744 Agomir reduced the degree of F-actin formation and cell proliferation while miR-744 Antagomir promoted these events, suggesting that miR-744 is a negative regulator of HSC activation.


MiR-744-led suppression in HSC activation is most likely through TGFβ1 because exogenous TGFβ1 nearly negated miR-744 Agomir’s action. This study suggests that reduction of miR-744 is a reliable biomarker for LC and miR-744/TGFβ1 relationship is a key regulator of LC.


Liver cirrhosis miRNA TGFβ1 Hepatic stellate cells miR-744 



Liver cirrhosis


Hepatic stellate cell


Extracellular matrix




Untranslated region


Transforming growth factor β


Epithelial-to-mesenchymal transition


Chronic hepatitis B


Quantitative reverse transcription polymerase chain reaction


Glyceraldehyde 3-phosphate dehydrogenase


Shanghai University of Traditional Chinese Medicine


Carbon tetrachloride


Phosphate buffered saline


Analysis of variance


Receiver–operator characteristic



This work is supported by National Natural Science Foundation of China (No. 81530101), NIH CA187152 and CA222467.

Author contributions

SR, SH and PL designed research. SR, JC, XL, WF and XZ performed the experiments and analyzed the data. YM, HZ, MS and CL contributed experimental materials or provided helpful suggestions. SR, JC, SH and PL wrote the manuscript.

Compliance with ethical standards

Conflict of interest

Shuang Ren, Jiamei Chen, Qinglan Wang, Xuewei Li, Ying Xu, Xiao Zhang, Yongping Mu, Hua Zhang, Shuang Huang, Ping Liu have no confict of interest to declare.

Ethics approval

Ethics approval all procedures performed in studies involving human participants and animals were in accordance with the ethical standards of Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine. The entire study was approved by Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine.

Informed consent

Informed consent was collected from patients to approve utilization of their samples for research purposes.

Supplementary material

12072_2019_9993_MOESM1_ESM.jpg (193 kb)
Supplementary material 1 (JPEG 193 kb)
12072_2019_9993_MOESM2_ESM.pdf (113 kb)
Supplementary material 2 (PDF 112 kb)


  1. 1.
    Jiao J, Friedman SL, Aloman C. Hepatic fibrosis. Curr Opin Gastroenterol 2009;25:223–229CrossRefGoogle Scholar
  2. 2.
    Friedman SL. Mechanisms of hepatic fibrogenesis. Gastroenterology 2008;134:1655–1669CrossRefGoogle Scholar
  3. 3.
    Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004;116:281–297CrossRefGoogle Scholar
  4. 4.
    Schueller F, Roy S, Vucur M, Trautwein C, Luedde T, Roderburg C. The role of miRNAs in the pathophysiology of liver diseases and toxicity. Int J Mol Sci 2018;19:261CrossRefGoogle Scholar
  5. 5.
    Ji J, Zhang J, Huang G, Qian J, Wang X, Mei S. Over-expressed microRNA-27a and 27b influence fat accumulation and cell proliferation during rat hepatic stellate cell activation. FEBS Lett 2009;583:759–766CrossRefGoogle Scholar
  6. 6.
    Sun X, He Y, Ma TT, Huang C, Zhang L, Li J. Participation of miR-200a in TGF-beta1-mediated hepatic stellate cell activation. Mol Cell Biochem 2014;388:11–23CrossRefGoogle Scholar
  7. 7.
    Zhang H, Li QY, Guo ZZ, Guan Y, Du J, Lu YY, et al. Serum levels of microRNAs can specifically predict liver injury of chronic hepatitis B. World J Gastroenterol 2012;18:5188–5196PubMedPubMedCentralGoogle Scholar
  8. 8.
    Iredale JP. Hepatic stellate cell behavior during resolution of liver injury. Semin Liver Dis 2001;21:427–436CrossRefGoogle Scholar
  9. 9.
    Gressner AM, Weiskirchen R, Breitkopf K, Dooley S. Roles of TGF-beta in hepatic fibrosis. Front Biosci 2002;7:d793–d807CrossRefGoogle Scholar
  10. 10.
    Shen H, Huang G, Hadi M, Choy P, Zhang M, Minuk GY, et al. Transforming growth factor-beta1 downregulation of Smad1 gene expression in rat hepatic stellate cells. Am J Physiol Gastrointest Liver Physiol 2003;285:G539–G546CrossRefGoogle Scholar
  11. 11.
    Cheng K, Yang N, Mahato RI. TGF-beta1 gene silencing for treating liver fibrosis. Mol Pharm 2009;6:772–779CrossRefGoogle Scholar
  12. 12.
    Qi Z, Atsuchi N, Ooshima A, Takeshita A, Ueno H. Blockade of type beta transforming growth factor signaling prevents liver fibrosis and dysfunction in the rat. Proc Natl Acad Sci USA 1999;96:2345–2349CrossRefGoogle Scholar
  13. 13.
    Chinese Society of Hepatology and Chinese Society of Infectious Diseases CMA. The guideline of prevention and treatment for chronic hepatitis B (2010 version). J Clin Hepatol 2011;27:I–XVIGoogle Scholar
  14. 14.
    Yi Y, Zhao Y, Li C, Zhang L, Huang H, Li Y, et al. RAID v2.0: an updated resource of RNA-associated interactions across organisms. Nucleic Acids Res 2017;45:D115–D118CrossRefGoogle Scholar
  15. 15.
    Blin K, Dieterich C, Wurmus R, Rajewsky N, Landthaler M, Akalin A. DoRiNA 2.0–upgrading the doRiNA database of RNA interactions in post-transcriptional regulation. Nucleic Acids Res 2015;43:D160–D167CrossRefGoogle Scholar
  16. 16.
    Bhattacharya A, Ziebarth JD, Cui Y. PolymiRTS Database 3.0: linking polymorphisms in microRNAs and their target sites with human diseases and biological pathways. Nucleic Acids Res 2014;42:D86–D91CrossRefGoogle Scholar
  17. 17.
    Xin X, Zhang Y, Liu X, Xin H, Cao Y, Geng M. MicroRNA in hepatic fibrosis and cirrhosis. Front Biosci (Landmark Ed). 2014;19:1418–1424CrossRefGoogle Scholar
  18. 18.
    Kwiecinski M, Noetel A, Elfimova N, Trebicka J, Schievenbusch S, Strack I, et al. Hepatocyte growth factor (HGF) inhibits collagen I and IV synthesis in hepatic stellate cells by miRNA-29 induction. PLoS One 2011;6:e24568CrossRefGoogle Scholar
  19. 19.
    Roderburg C, Urban GW, Bettermann K, Vucur M, Zimmermann H, Schmidt S, et al. Micro-RNA profiling reveals a role for miR-29 in human and murine liver fibrosis. Hepatology 2011;53:209–218CrossRefGoogle Scholar
  20. 20.
    Cushing L, Kuang PP, Qian J, Shao F, Wu J, Little F, et al. miR-29 is a major regulator of genes associated with pulmonary fibrosis. Am J Respir Cell Mol Biol 2011;45:287–294CrossRefGoogle Scholar
  21. 21.
    Song MY, Pan KF, Su HJ, Zhang L, Ma JL, Li JY, et al. Identification of serum microRNAs as novel non-invasive biomarkers for early detection of gastric cancer. PLoS One 2012;7:e33608CrossRefGoogle Scholar
  22. 22.
    Kubiczkova L, Kryukov F, Slaby O, Dementyeva E, Jarkovsky J, Nekvindova J, et al. Circulating serum microRNAs as novel diagnostic and prognostic biomarkers for multiple myeloma and monoclonal gammopathy of undetermined significance. Haematologica 2014;99:511–518CrossRefGoogle Scholar
  23. 23.
    Mi QS, Weiland M, Qi RQ, Gao XH, Poisson LM, Zhou L. Identification of mouse serum miRNA endogenous references by global gene expression profiles. PLoS One 2012;7:e31278CrossRefGoogle Scholar
  24. 24.
    Blobe GC, Schiemann WP, Lodish HF. Role of transforming growth factor beta in human disease. N Engl J Med 2000;342:1350–1358CrossRefGoogle Scholar
  25. 25.
    Martin J, Jenkins RH, Bennagi R, Krupa A, Phillips AO, Bowen T, et al. Post-transcriptional regulation of Transforming Growth Factor Beta-1 by microRNA-744. PLoS One 2011;6:e25044CrossRefGoogle Scholar
  26. 26.
    Matsuzaki K, Murata M, Yoshida K, Sekimoto G, Uemura Y, Sakaida N, et al. Chronic inflammation associated with hepatitis C virus infection perturbs hepatic transforming growth factor beta signaling, promoting cirrhosis and hepatocellular carcinoma. Hepatology 2007;46:48–57CrossRefGoogle Scholar
  27. 27.
    Zhang CY, Yuan WG, He P, Lei JH, Wang CX. Liver fibrosis and hepatic stellate cells: etiology, pathological hallmarks and therapeutic targets. World J Gastroenterol 2016;22:10512–10522CrossRefGoogle Scholar
  28. 28.
    Guo CJ, Pan Q, Li DG, Sun H, Liu BW. miR-15b and miR-16 are implicated in activation of the rat hepatic stellate cell: an essential role for apoptosis. J Hepatol 2009;50:766–778CrossRefGoogle Scholar
  29. 29.
    Zheng J, Lin Z, Dong P, Lu Z, Gao S, Chen X, et al. Activation of hepatic stellate cells is suppressed by microRNA-150. Int J Mol Med 2013;32:17–24CrossRefGoogle Scholar
  30. 30.
    Li F, Ma N, Zhao R, Wu G, Zhang Y, Qiao Y, et al. Overexpression of miR-483-5p/3p cooperate to inhibit mouse liver fibrosis by suppressing the TGF-beta stimulated HSCs in transgenic mice. J Cell Mol Med 2014;18:966–974CrossRefGoogle Scholar

Copyright information

© Asian Pacific Association for the Study of the Liver 2019

Authors and Affiliations

  1. 1.Key Laboratory of Liver and Kidney Diseases (Ministry of Education)Institute of Liver Diseases, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese MedicineShanghaiChina
  2. 2.Traditional Chinese Medicine DepartmentFirst Affiliated Hospital of China Medical UniversityShenyangChina
  3. 3.Department of Anatomy and Cell BiologyUniversity of Florida College of MedicineGainesvilleUSA

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