Overexpression of ring finger protein 20 inhibits the progression of liver fibrosis via mediation of histone H2B lysine 120 ubiquitination

Abstract

Liver fibrosis is a chronic liver injury that leads to liver cirrhosis and liver cancer. Ring finger protein 20 (RNF20), also named as E3 ubiquitin-protein ligase BRE1A, has been reported to be involved in chronic liver diseases. However, the role of RNF20 in liver fibrosis remains unclear. To mimic liver fibrosis in vitro, LX-2 cells were treated with TGF-β. Gene and protein expressions were detected by RT-qPCR and western blot, respectively. The mechanism by which RNF20 mediated the progression of liver fibrosis was explored by bioinformatics analysis. Finally, in vivo mouse model of liver fibrosis was established to detect the function of RNF20. The results indicated that TGF-β-induced increase of cell viability and migration was significantly reversed by RNF20 overexpression. Consistently, overexpression of RNF20 significantly reversed TGF-β-induced activation of fibrotic proteins in LX-2 cells. Meanwhile, VEGFA, TNF-α and IL-6 were found to be the downstream targets of RNF20 in LX-2 cells. Moreover, RNF20 overexpression notably inhibited the progression of liver fibrosis via ubiquitination of H2B. Finally, RNF20 upregulation significantly attenuated the symptom of liver fibrosis in vivo. In summary, overexpression of RNF20 significantly inhibited the progression of liver fibrosis in vitro and in vivo. Therefore, RNF20 might serve as a new target for the treatment of liver fibrosis.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

References

  1. 1.

    Manning MW, Li YJ, Linder D, Haney JC, Wu YH, Podgoreanu MV, et al. (2020). Conventional Ultrafiltration During Elective Cardiac Surgery and Postoperative Acute Kidney Injury. J Cardiothorac Vasc Anesth S1053-0770:31271-4.

  2. 2.

    Gheorghe G, Bungau S, Ceobanu G, Ilie M, Bacalbasa N, Bratu OG, et al. The non-invasive assessment of hepatic fibrosis. J Formos Med Assoc. 2020;120:794–803.

    Article  Google Scholar 

  3. 3.

    Hukkinen M, Pihlajoki M, Pakarinen MP. Predicting native liver injury and survival in biliary atresia. Semin Pediatr Surg. 2020;29:150943.

    Article  Google Scholar 

  4. 4.

    Mellinger J, Winder GS, Fernandez AC (2020). HEP-20–1087: Measuring the Alcohol in Alcohol-related Liver Disease: Choices and Challenges for Clinical Research. Hepatology (Online ahead of print).

  5. 5.

    Liang X, Tao C, Pan J, Zhang L, Liu L, Zhao Y, et al. (2020). Rnf20 deficiency in adipocyte impairs adipose tissue development and thermogenesis. Protein Cell (Online ahead of print).

  6. 6.

    Zhao Y, Yang S, Wang Y, Tao C. Molecular characterization, expression profiling, and SNP analysis of the porcine RNF20 gene. Animals (Basel). 2020;10:888.

    Article  Google Scholar 

  7. 7.

    Manikoth Ayyathan D, Koganti P, Marcu-Malina V, Litmanovitch T, Trakhtenbrot L, Emanuelli A, et al. SMURF2 prevents detrimental changes to chromatin, protecting human dermal fibroblasts from chromosomal instability and tumorigenesis. Oncogene. 2020;39:3396–410.

    CAS  Article  Google Scholar 

  8. 8.

    Jeon YG, Lee JH, Ji Y, Sohn JH, Lee D, Kim DW, et al. RNF20 functions as a transcriptional coactivator for PPARgamma by promoting NCoR1 degradation in adipocytes. Diabetes. 2020;69:20–34.

    CAS  Article  Google Scholar 

  9. 9.

    Robson A, Makova SZ, Barish S, Zaidi S, Mehta S, Drozd J, et al. Histone H2B monoubiquitination regulates heart development via epigenetic control of cilia motility. Proc Natl Acad Sci USA. 2019;116:14049–54.

    CAS  Article  Google Scholar 

  10. 10.

    Wu C, Cui Y, Liu X, Zhang F, Lu LY, Yu X. The RNF20/40 complex regulates p53-dependent gene transcription and mRNA splicing. J Mol Cell Biol. 2020;12:113–24.

    CAS  Article  Google Scholar 

  11. 11.

    Hooda J, Novak M, Salomon MP, Matsuba C, Ramos RI, MacDuffie E, et al. Early loss of histone H2B monoubiquitylation alters chromatin accessibility and activates key immune pathways that facilitate progression of ovarian cancer. Cancer Res. 2019;79:760–72.

    CAS  Article  Google Scholar 

  12. 12.

    Oh S, Boo K, Kim J, Baek SA, Jeon Y, You J, et al. The chromatin-binding protein PHF6 functions as an E3 ubiquitin ligase of H2BK120 via H2BK12Ac recognition for activation of trophectodermal genes. Nucleic Acids Res. 2020;48:9037.

    Article  Google Scholar 

  13. 13.

    Valencia-Sanchez MI, De Ioannes P, Wang M, Vasilyev N, Chen R, Nudler E, et al. Structural basis of dot1l stimulation by histone H2B lysine 120 ubiquitination. Mol Cell. 2019;74(1010–9):e6.

    Google Scholar 

  14. 14.

    Wojcik F, Dann GP, Beh LY, Debelouchina GT, Hofmann R, Muir TW. Functional crosstalk between histone H2B ubiquitylation and H2A modifications and variants. Nat Commun. 2018;9:1394.

    Article  Google Scholar 

  15. 15.

    Desmet VJ, Gerber M, Hoofnagle JH, Manns M, Scheuer PJ. Classification of chronic hepatitis: diagnosis, grading and staging. Hepatology. 1994;19:1513–20.

    CAS  Article  Google Scholar 

  16. 16.

    Jia B, Zhang S, Wu S, Zhu Q, Li W. MiR-770 promotes oral squamous cell carcinoma migration and invasion by regulating the Sirt7/Smad4 pathway. IUBMB Life. 2020;73:264.

    Article  Google Scholar 

  17. 17.

    Zhang L, Liu X, Liang J, Wu J, Tan D, Hu W. Lefty-1 inhibits renal epithelial-mesenchymal transition by antagonizing the TGF-beta/Smad signaling pathway. J Mol Histol. 2020;51:77–87.

    CAS  Article  Google Scholar 

  18. 18.

    Zhu H, Shan Y, Ge K, Lu J, Kong W, Jia C. Specific overexpression of mitofusin-2 in hepatic stellate cells ameliorates liver fibrosis in mice model. Hum Gene Ther. 2020;31:103–9.

    CAS  Article  Google Scholar 

  19. 19.

    Elnfarawy AA, Nashy AE, Abozaid AM, Komber IF, Elweshahy RH, Abdelrahman RS. Vinpocetine attenuates thioacetamide-induced liver fibrosis in rats. Hum Exp Toxicol. 2020;2020:960327120947453.

    Google Scholar 

  20. 20.

    Sheng J, Zhang B, Chen Y, Yu F. Capsaicin attenuates liver fibrosis by targeting Notch signaling to inhibit TNF-alpha secretion from M1 macrophages. Immunopharmacol Immunotoxicol. 2020;42:1–10.

    Article  Google Scholar 

  21. 21.

    Xu Z, Song Z, Li G, Tu H, Liu W, Liu Y, et al. H2B ubiquitination regulates meiotic recombination by promoting chromatin relaxation. Nucleic Acids Res. 2016;44:9681–97.

    CAS  Article  Google Scholar 

  22. 22.

    Tarcic O, Pateras IS, Cooks T, Shema E, Kanterman J, Ashkenazi H, et al. RNF20 links histone H2B ubiquitylation with inflammation and inflammation-associated cancer. Cell Rep. 2016;14:1462–76.

    CAS  Article  Google Scholar 

  23. 23.

    Chernikova SB, Razorenova OV, Higgins JP, Sishc BJ, Nicolau M, Dorth JA, et al. Deficiency in mammalian histone H2B ubiquitin ligase Bre1 (Rnf20/Rnf40) leads to replication stress and chromosomal instability. Cancer Res. 2012;72:2111–9.

    CAS  Article  Google Scholar 

  24. 24.

    Vethantham V, Yang Y, Bowman C, Asp P, Lee JH, Skalnik DG, et al. Dynamic loss of H2B ubiquitylation without corresponding changes in H3K4 trimethylation during myogenic differentiation. Mol Cell Biol. 2012;32:1044–55.

    CAS  Article  Google Scholar 

  25. 25.

    de Oliveira A, Castanhole-Nunes MMU, Biselli-Chicote PM, Pavarino EC, da Silva R, da Silva RF, et al. Differential expression of angiogenesis-related miRNAs and VEGFA in cirrhosis and hepatocellular carcinoma. Arch Med Sci. 2020;16:1150–7.

    Article  Google Scholar 

  26. 26.

    Lee KT, Islam F, Vider J, Martin J, Chruscik A, Lu CT, et al. Overexpression of family with sequence similarity 134, member B (FAM134B) in colon cancers and its tumor suppressive properties in vitro. Cancer Biol Ther. 2020;21:1–9.

    Article  Google Scholar 

  27. 27.

    Gong H, Fan Z, Yi D, Chen J, Li Z, Guo R, et al. Histidine kinase NME1 and NME2 are involved in TGF-beta1-induced HSC activation and CCl4-induced liver fibrosis. J Mol Histol. 2020;51:573.

    CAS  Article  Google Scholar 

  28. 28.

    Xue Y, Fan X, Yang R, Jiao Y, Li Y. miR-29b-3p inhibits post-infarct cardiac fibrosis by targeting FOS. Biosci Rep. 2020; 40:BSR20201227.

    CAS  Article  Google Scholar 

  29. 29.

    Song XH, Chen XZ, Chen XL, Liu K, Zhang WH, Mo XM, et al. Peritoneal metastatic cancer stem cells of gastric cancer with partial mesenchymal-epithelial transition and enhanced invasiveness in an intraperitoneal transplantation model. Gastroenterol Res Pract. 2020;2020:3256538.

    Article  Google Scholar 

Download references

Funding

The study was supported by the National Natural Science Foundation of China (No. 81672115).

Author information

Affiliations

Authors

Contributions

HH design study and revised the manuscript; SC and XD analysis data and perform manuscript drafting; HL, YG and YZ search the literature and collect data; HH reviewed the results and made critical comments on the manuscript.

Corresponding author

Correspondence to Haijun Huang.

Ethics declarations

Conflict of interests

These authors declared no competing interests in this research.

Ethical approval

In vivo study, all the protocols were performed according to the guidelines of the Use Committee of Zhejiang Provincial People’s Hospital (No. 20200004), and all the animal experiments were performed in accordance with National Institutes of Health guide for the care and use of laboratory animals. The clinical samples were collected in accordance with the Institutional Ethical Committee of Zhejiang Provincial People’s Hospital.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Chen, S., Dai, X., Li, H. et al. Overexpression of ring finger protein 20 inhibits the progression of liver fibrosis via mediation of histone H2B lysine 120 ubiquitination. Human Cell (2021). https://doi.org/10.1007/s13577-021-00498-z

Download citation

Keywords

  • Liver fibrosis
  • RNF20
  • H2BK120ub
  • Ubiquitination