Skip to main content

Advertisement

SpringerLink
Log in

Integration of hepatitis B virus DNA into p21-activated kinase 3 (PAK3) gene in HepG2.2.15 cells

  • Original Paper
  • Published:
Virus Genes Aims and scope Submit manuscript

Abstract

Integration of HBV DNA into host chromosomes was found in most of the patients with chronic hepatitis B (CHB). In this study, using inverse nested PCR (invPCR), we found the integration site chrX: 111,009,033, which inserted into the p21-activated kinase 3 (PAK3) gene in HepG2.2.15 cells. The viral-human chimeric transcripts were also observed and, significant differences of the copy numbers of integration site chrX: 111,009,033 (P = 0.012) and intra-cell HBV DNA levels (P = 0.027) were found between the cells with and without H2O2 treatment, respectively. This study may provide a novel insight into the elucidation of etiology of HBV integration.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Ott JJ, Stevens GA, Groeger J, Wiersma ST (2012) Global epidemiology of hepatitis B virus infection: new estimates of age-specific HBsAg seroprevalence and endemicity. Vaccine 30:2212–2219

    Article  CAS  Google Scholar 

  2. Trepo C, Chan HL, Lok A (2014) Hepatitis B virus infection. Lancet 384:2053–2063

    Article  CAS  Google Scholar 

  3. Yan HL, Yang Y, Zhang L, Tang GN, Wang YZ, Xue G, Zhou WP, Sun SH (2015) Characterization of the genotype and integration patterns of hepatitis B virus in early- and late-onset hepatocellular carcinoma. Hepatology 61:1821–1831

    Article  CAS  Google Scholar 

  4. Sung WK, Zheng H, Li S, Chen R, Liu X, Li Y, Lee NP, Lee WH, Ariyaratne PN, Tennakoon C et al (2012) Genome-wide survey of recurrent HBV integration in hepatocellular carcinoma. Nat Genet 44:765–769

    Article  CAS  Google Scholar 

  5. Wang HC, Huang W, Lai MD, Su IJ (2006) Hepatitis B virus pre-S mutants, endoplasmic reticulum stress and hepatocarcinogenesis. Cancer Sci 97:683–688

    Article  CAS  Google Scholar 

  6. Tu T, Budzinska MA, Shackel NA, Jilbert AR (2015) Conceptual models for the initiation of hepatitis B virus-associated hepatocellular carcinoma. Liver Int 35:1786–1800

    Article  CAS  Google Scholar 

  7. Murakami Y, Saigo K, Takashima H, Minami M, Okanoue T, Bréchot C, Paterlini-Bréchotet P (2005) Large scaled analysis of hepatitis B virus (HBV) DNA integration in HBV related hepatocellular carcinomas. Gut 54:1162–1168

    Article  CAS  Google Scholar 

  8. Saigo K, Yoshida K, Ikeda R, Sakamoto Y, Murakami Y, Urashima T, Asano T, Kenmochi T, Inoue I (2008) Integration of hepatitis B virus DNA into the myeloid/lymphoid or mixedlineage leukemia (MLL4) gene and rearrangements of MLL4 in human hepatocellular carcinoma. Hum Mutat 29:703–708

    Article  CAS  Google Scholar 

  9. Tamori A, Yamanishi Y, Kawashima S, Kanehisa M, Enomoto M, Tanaka H, Kubo S, Shiomi S, Nishiguchi S (2005) Alteration of gene expression in human hepatocellular carcinoma with integrated hepatitis B virus DNA. Clin Cancer Res 11:5821–5826

    Article  CAS  Google Scholar 

  10. Ding D, Lou X, Hua D, Yu W, Li L, Wang J, Gao F, Zhao N, Ren GP, Li LJ et al (2012) Recurrent targeted genes of hepatitis B virus in the liver cancer genomes identified by a next-generation sequencing-based approach. PLoS Genet 8:e1003065

    Article  CAS  Google Scholar 

  11. Lau CC, Sun T, Ching AK, He M, Li JW, Wong AM, Co NN, Chan AW, Li PS, Lung RW et al (2014) Viral-human chimeric transcript predisposes risk to liver cancer development and progression. Cancer Cell 25:335–349

    Article  CAS  Google Scholar 

  12. Petersen J, Dandri M, Bürkle A, Zhang L, Rogler CE (1997) Increase in the frequency of hepadnavirus DNA integrations by oxidative DNA damage and inhibition of DNA repair. J Virol 71:5455–5463

    Article  CAS  Google Scholar 

  13. Tu T, Jilbert AR (2017) Detection of Hepatocyte clones containing integrated hepatitis B virus DNA using inverse nested PCR. Methods Mol Biol 1540:97–118

    Article  CAS  Google Scholar 

  14. Ruan P, Dai XF, Sun J, He CP, Huang C, Zhou R, Cao Z, Ye L (2019) Different types of viral-host junction found in HBV integration breakpoints in HBV-infected patients. Mol Med Rep 19:1410–1416

    CAS  PubMed  Google Scholar 

  15. Werle-Lapostolle B, Bowden S, Locarnini S, Wursthorn K, Petersen J, Lau G, Trepo C, Marcellin P, Goodman Z, Delaney WE IV (2004) Persistence of cccDNA during the natural history of chronic hepatitis B and decline during adefovir dipivoxil therapy. Gastroenterology 126:1750–1758

    Article  CAS  Google Scholar 

  16. Tu T, Budzinska MA, Vondran FWR, Shackel NA, Urban S (2018) Hepatitis B virus DNA integration occurs early in the viral life cycle in an in vitro infection model via NTCP-dependent uptake of enveloped virus particles. J Virol. https://doi.org/10.1128/JVI.02007-17

    Article  PubMed  PubMed Central  Google Scholar 

  17. Minami M, Poussin K, Brechot C, Paterlini P (1995) A novel PCR technique using Alu-specific primers to identify unknown flanking sequences from the human genome. Genomics 29:403–408

    Article  CAS  Google Scholar 

  18. Jiang Z, Jhunjhunwala S, Liu J, Haverty PM, Kennemer MI, Guan Y, Lee W, Carnevali P, Stinson J, Johnson S et al (2012) The effects of hepatitis B virus integration into the genomes of hepatocellular carcinoma patients. Genome Res 22:593–601

    Article  CAS  Google Scholar 

  19. Toh ST, Jin Y, Liu L, Wang J, Babrzadeh F, Gharizadeh B, Ronaghi M, Toh HC, Chow PK, Chung AY et al (2013) Deep sequencing of the hepatitis B virus in hepatocellular carcinoma patients reveals enriched integration events, structural alterations and sequence variations. Carcinogenesis 34:787–798

    Article  CAS  Google Scholar 

  20. Zhao LH, Liu X, Yan HX, Li WY, Zeng X, Yang Y, Zhao J, Liu SP, Zhuang XH, Lin C et al (2016) Genomic and oncogenic preference of HBV integration in hepatocellular carcinoma. Nat Commun 7:12992

    Article  CAS  Google Scholar 

  21. Li WY, Zeng X, Lee N et al (2013) HIVID: An efficient method to detect HBV integration using low coverage sequencing. Genomics 102:338–344

    Article  CAS  Google Scholar 

  22. Boda B, Alberi S, Nikonenko I, Node-Langlois R, Jourdain P, Moosmayer M, Parisi-Jourdain L, Muller D (2004) The mental retardation protein PAK3 contributes to synapse formation and plasticity in hippocampus. J Neuro Sci 24:10816–10825

    CAS  Google Scholar 

  23. Zhou NN, Ding B, Agler M, Cockett M, McPhee F (2005) Lethality of PAK3 and SGK2 shRNAs to human papillomavirus positive cervical cancer cells is independent of PAK3 and SGK2 knockdown. PLoS ONE 10:e0117357

    Article  Google Scholar 

  24. Bienvenu T, desPortes V, McDonell N, Carrié A, Zemni R, Couvert P, Ropers HH, Moraine C, van Bokhoven H, Fryns JP, et al (2000) Missense mutation in PAK3, R67C, causes X-linked non specific mental retardation. Am J Med Genet 93:294–298.

    Article  CAS  Google Scholar 

  25. Li X, Zhang J, Yang Z, Kang J, Jiang S, Zhang T, Chen T, Li M, Lv Q, Chen X et al (2014) The function of targeted host genes determines the oncogenicity of HBV integration in hepatocellular carcinoma. J Hepatol 60:975–984

    Article  Google Scholar 

  26. Dandri M, Burda MR, Bürkle A, Zuckerman DM, Will H, Rogler CE, Greten H, Petersen J (2002) Increase in de novo HBV DNA integrations in response to oxidative DNA damage or inhi- bition of poly(ADP-ribosyl)ation. Hepatology 35:217–223

    Article  CAS  Google Scholar 

  27. Bill CA, Summers J (2004) Genomic DNA double-strand breaks are targets for hepadnaviral DNA integration. Proc Natl Acad Sci USA 101:11135–11140

    Article  CAS  Google Scholar 

  28. Hu Z, Zhu D, Wang W, Li W, Jia W, Zeng X, Ding W, Yu L, Wang X, Wang L et al (2015) Genome-wide profiling of HPV integration in cervical cancer identifies clustered genomic hot spots and a potential microhomology-mediated integration mechanism. Nat Genet 47:158–163

    Article  CAS  Google Scholar 

  29. Mladenov E, Magin S, Soni A, Iliakis G (2016) DNA double-strand-break repair in higher eukaryotes and its role in genomic instability and cancer: cell cycle and proliferation-dependent regulation. Semin Cancer Biol 37–38:51–64

    Article  Google Scholar 

  30. Yang L, Ye S, Zhao X, Ji L, Zhang Y, Zhou PY, Sun J, Guan YF, Han YX, Ni C et al (2018) Molecular characterization of HBV DNA integration in patients with hepatitis and hepatocellular carcinoma. J Cancer 9:3225–3235

    Article  Google Scholar 

  31. Schraufstatter IU, Hyslop PA, Hinshaw DB, Spragg RG, Sklar LA, Cochrane CG (1986) Hydrogen peroxide-induced injury of cells and its prevention by poly (ADP-ribose) polymerase. Proc Natl Acad Sci USA 83:4908–4912

    Article  CAS  Google Scholar 

  32. Schraufstatter IU, Hinshaw DB, Hyslop PA, Spragg RG, Cochrane CG (1986) Oxidant injury of cells. J Clin Invest 77:1312–1320

    Article  CAS  Google Scholar 

  33. Mason WS, Jilbert AR, Summers J (2005) Clonal expansion of hepatocytes during chronic woodchuck hepatitis virus infection. Proc Natl Acad Sci USA 102:1139–1144

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Mrs. Francoise BERBY and Anaelle DUBOIS from Centre de Recherche en Cancérologie, Université Claude Bernard, Lyon for their technical assistance.

Funding

This study was funded by China Scholarship Council (Grant No. 201706275092).

Author information

Authors and Affiliations

  1. Department of Gastroenterology, Renmin Hospital of Wuhan University, 238 Jiefang Road, Wuhan, 430060, Hubei, People’s Republic of China

    Peng Ruan, Jun Sun, Chunping He, Chao Huang & Rui Zhou

  2. Department of Breast Surgery, Renmin Hospital, Hubei University of Medicine, Shiyan, 442000, Hubei, People’s Republic of China

    Peng Ruan & Xiufang Dai

  3. INSERM U1052, CNRS UMR5286, Centre de Recherche en Cancérologie, Université Claude Bernard, Lyon, France

    Peng Ruan & Isabelle Chemin

Authors
  1. Peng Ruan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiufang Dai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jun Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chunping He
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chao Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Rui Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Isabelle Chemin
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

PR and IC conceived and designed the experiments. PR, XFD and JC performed the experiments. CH designed all the primers. RZ modified the English language of the manuscript. CPH analyzed the data. PR wrote the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Peng Ruan or Isabelle Chemin.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interests.

Research involving human and/or animals participants

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Edited by Zhen F. Fu.

Publisher's Note

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

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ruan, P., Dai, X., Sun, J. et al. Integration of hepatitis B virus DNA into p21-activated kinase 3 (PAK3) gene in HepG2.2.15 cells. Virus Genes 56, 168–173 (2020). https://doi.org/10.1007/s11262-019-01725-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11262-019-01725-4

Keywords

Search

Navigation