Skip to main content

Advertisement

SpringerLink
Log in

Comprehensive analyses of mutations and hepatitis B virus integration in hepatocellular carcinoma with clinicopathological features

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Background and Aims

Genetic alterations in specific genes are critical events in carcinogenesis and hepatocellular carcinoma (HCC) progression. However, the genetic alterations responsible for HCC development, progression, and survival are unclear.

Methods

We investigated the essential difference in genetic alterations between HCC and adjacent non-HCC tissues using next-generation sequencing technology.

Results

We found recurrent mutations in several genes such as telomerase reverse transcriptase (TERT; 65 % of the total 104 HCCs), TP53 (38 %), CTNNB1 (30 %), AXIN1 (2 %), PTEN (2 %), and CDKN2A (2 %). TERT promoter mutations were associated with older age (p = 0.005), presence of hepatitis C virus (HCV) infection (p = 0.003), and absence of hepatitis B virus (HBV) infection (p < 0.0001). In hepatitis B surface antigen (HBs Ag)-positive HCC without TERT promoter mutations, HBV integration into TERT locus was found in 47 % patients and was mutually exclusive to TERT promoter mutations. Most (89 %) HBV integrants were in the HBx region. TP53 mutations were associated with HBV infection (p = 0.0001) and absence of HCV infection (p = 0.002). CTNNB1 mutations were associated with absence of HBV infection (p = 0.010). Moreover, TERT promoter mutation was significantly associated with shorter disease-free survival (p = 0.005) and poor overall survival (p = 0.024).

Conclusions

Gene alterations in TERT promoter, TP53, CTNNB1, and HBV integration were closely associated with HCC development, and mutations in TERT promoter are related to poor prognosis. These results are useful for understanding the underlying mechanism of hepatocarcinogenesis, diagnosis, and predicting outcomes of patients with HCC.

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

Similar content being viewed by others

Abbreviations

HCC:

Hepatocellular carcinoma

TERT :

Telomerase reverse transcriptase

HCV:

Hepatitis C virus

HBV:

Hepatitis B virus

HBsAg:

Hepatitis B surface antigen

PCR:

Polymerase chain reaction

ISPs:

Ion sphere particles

COSMIC:

Catalogue of somatic mutations in cancer

gDNA:

Genomic DNA

CT:

Computed tomography

MRI:

Magnetic resonance imaging

AFP:

Alpha-fetoprotein

DFS:

Disease-free survival

OS:

Overall survival

Anti-HBc:

Antibody to hepatitis B core antigen

Anti-HBs:

Antibody to hepatitis B surface antigen

References

  1. El-Serag HB. Epidemiology of viral hepatitis and hepatocellular carcinoma. Gastroenterology. 2012;142(1264–73):e1.

    PubMed  Google Scholar 

  2. Thorgeirsson SS, Grisham JW. Molecular pathogenesis of human hepatocellular carcinoma. Nat Genet. 2002;31:339–46.

    Article  CAS  PubMed  Google Scholar 

  3. Huang J, Deng Q, Wang Q, et al. Exome sequencing of hepatitis B virus-associated hepatocellular carcinoma. Nat Genet. 2012;44:1117–21.

    Article  CAS  PubMed  Google Scholar 

  4. Guichard C, Amaddeo G, Imbeaud S, et al. Integrated analysis of somatic mutations and focal copy-number changes identifies key genes and pathways in hepatocellular carcinoma. Nat Genet. 2012;44:694–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Fujimoto A, Totoki Y, Abe T, et al. Whole-genome sequencing of liver cancers identifies etiological influences on mutation patterns and recurrent mutations in chromatin regulators. Nat Genet. 2012;44:760–4.

    Article  CAS  PubMed  Google Scholar 

  6. Li M, Zhao H, Zhang X, et al. Inactivating mutations of the chromatin remodeling gene ARID2 in hepatocellular carcinoma. Nat Genet. 2011;43:828–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Killela PJ, Reitman ZJ, Jiao Y, et al. TERT promoter mutations occur frequently in gliomas and a subset of tumors derived from cells with low rates of self-renewal. Proc Natl Acad Sci USA. 2013;110:6021–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Nault JC, Mallet M, Pilati C, et al. High frequency of telomerase reverse-transcriptase promoter somatic mutations in hepatocellular carcinoma and preneoplastic lesions. Nat Commun. 2013;4:2218.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Huang FW, Hodis E, Xu MJ, Kryukov GV, Chin L, Garraway LA. Highly recurrent TERT promoter mutations in human melanoma. Science. 2013;339:957–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Horn S, Figl A, Rachakonda PS, et al. TERT promoter mutations in familial and sporadic melanoma. Science. 2013;339:959–61.

    Article  CAS  PubMed  Google Scholar 

  11. Beadling C, Neff TL, Heinrich MC, et al. Combining highly multiplexed PCR with semiconductor-based sequencing for rapid cancer genotyping. J Mol Diagn. 2013;15:171–6.

    Article  CAS  PubMed  Google Scholar 

  12. Takano S, Fukasawa M, Maekawa S, et al. Deep sequencing of cancer-related genes revealed GNAS mutations to be associated with intraductal papillary mucinous neoplasms and its main pancreatic duct dilation. PLoS One. 2014;9:e98718.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Forbes SA, Bindal N, Bamford S, et al. COSMIC: mining complete cancer genomes in the catalogue of somatic mutations in cancer. Nucleic Acids Res. 2011;39:D945–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Jiang Z, Jhunjhunwala S, Liu J, et al. The effects of hepatitis B virus integration into the genomes of hepatocellular carcinoma patients. Genome Res. 2012;22:593–601.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Sung WK, Zheng H, Li S, et al. Genome-wide survey of recurrent HBV integration in hepatocellular carcinoma. Nat Genet. 2012;44:765–9.

    Article  CAS  PubMed  Google Scholar 

  16. Li X, Zhang J, Yang Z, et al. The function of targeted host genes determines the oncogenicity of HBV integration in hepatocellular carcinoma. J Hepatol. 2014;60:975–84.

    Article  PubMed  Google Scholar 

  17. Chen YL, Jeng YM, Chang CN, et al. TERT promoter mutation in resectable hepatocellular carcinomas: a strong association with hepatitis C infection and absence of hepatitis B infection. Int J Surg. 2014;12:659–65.

    Article  CAS  PubMed  Google Scholar 

  18. Nault JC, Calderaro J, Tommaso LD, et al. Telomerase reverse transcriptase promoter mutation is an early somatic genetic alteration in the transformation of premalignant nodules in hepatocellular carcinoma on cirrhosis. Hepatology. 2014;60:1983–92.

    Article  CAS  PubMed  Google Scholar 

  19. Ng J, Wu J. Hepatitis B- and hepatitis C-related hepatocellular carcinomas in the United States: similarities and differences. Hepat Mon. 2012;12:e7635.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Sasaki Y, Yamada T, Tanaka H, et al. Risk of recurrence in a long-term follow-up after surgery in 417 patients with hepatitis B- or hepatitis C-related hepatocellular carcinoma. Ann Surg. 2006;244:771–80.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Minami T, Tateishi R, Shiina S, et al. Comparison of improved prognosis between hepatitis B- and hepatitis C-related hepatocellular carcinoma. Hepatol Res. 2015;45:E99–107.

    Article  CAS  PubMed  Google Scholar 

  22. Takenaka K, Yamamoto K, Taketomi A, et al. A comparison of the surgical results in patients with hepatitis B versus hepatitis C-related hepatocellular carcinoma. Hepatology. 1995;22:20–4.

    CAS  PubMed  Google Scholar 

  23. Paterlini-Bréchot P, Saigo K, Murakami Y, et al. Hepatitis B virus-related insertional mutagenesis occurs frequently in human liver cancers and recurrently targets human telomerase gene. Oncogene. 2003;22:3911–6.

    Article  PubMed  Google Scholar 

  24. Toh ST, Jin Y, Liu L, et al. Deep sequencing of the hepatitis B virus in hepatocellular carcinoma patients reveals enriched integration events, structural alterations and sequence variations. Carcinogenesis. 2013;34:787–98.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Author notes

    Authors and Affiliations

    1. Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan

      Fukiko Kawai-Kitahata, Yasuhiro Asahina, Sei Kakinuma, Miyako Murakawa, Sayuri Nitta, Takako Watanabe, Satoshi Otani, Miki Taniguchi, Fumio Goto, Hiroko Nagata, Shun Kaneko, Megumi Tasaka-Fujita, Yuki Nishimura-Sakurai, Seishin Azuma, Yasuhiro Itsui, Mina Nakagawa & Mamoru Watanabe

    2. Department of Liver Disease Control, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan

      Yasuhiro Asahina & Sei Kakinuma

    3. Department of Molecular Oncology, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan

      Shinji Tanaka

    4. Department of Hepato-Biliary-Pancreatic Surgery, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan

      Minoru Tanabe

    5. First Department of Internal Medicine, Faculty of Medicine, University of Yamanashi, 1110 Shimogato, Chuo-shi, Yamanshi, 409-3898, Japan

      Shinichi Takano, Mitsuharu Fukasawa, Minoru Sakamoto, Shinya Maekawa & Nobuyuki Enomoto

    Authors
    1. Fukiko Kawai-Kitahata
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Yasuhiro Asahina
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Shinji Tanaka
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Sei Kakinuma
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Miyako Murakawa
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Sayuri Nitta
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Takako Watanabe
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Satoshi Otani
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. Miki Taniguchi
      View author publications

      You can also search for this author in PubMed Google Scholar

    10. Fumio Goto
      View author publications

      You can also search for this author in PubMed Google Scholar

    11. Hiroko Nagata
      View author publications

      You can also search for this author in PubMed Google Scholar

    12. Shun Kaneko
      View author publications

      You can also search for this author in PubMed Google Scholar

    13. Megumi Tasaka-Fujita
      View author publications

      You can also search for this author in PubMed Google Scholar

    14. Yuki Nishimura-Sakurai
      View author publications

      You can also search for this author in PubMed Google Scholar

    15. Seishin Azuma
      View author publications

      You can also search for this author in PubMed Google Scholar

    16. Yasuhiro Itsui
      View author publications

      You can also search for this author in PubMed Google Scholar

    17. Mina Nakagawa
      View author publications

      You can also search for this author in PubMed Google Scholar

    18. Minoru Tanabe
      View author publications

      You can also search for this author in PubMed Google Scholar

    19. Shinichi Takano
      View author publications

      You can also search for this author in PubMed Google Scholar

    20. Mitsuharu Fukasawa
      View author publications

      You can also search for this author in PubMed Google Scholar

    21. Minoru Sakamoto
      View author publications

      You can also search for this author in PubMed Google Scholar

    22. Shinya Maekawa
      View author publications

      You can also search for this author in PubMed Google Scholar

    23. Nobuyuki Enomoto
      View author publications

      You can also search for this author in PubMed Google Scholar

    24. Mamoru Watanabe
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Yasuhiro Asahina.

    Ethics declarations

    Conflict of interest

    Dr. Asahina and Dr. Kakinuma belong to a donation-funded department funded by Chugai Pharmaceutical Co. Ltd., Toray Industries Inc., Bristol-Myers Squibb, Dainippon Sumitomo Pharma Co. Ltd., and Merck Sharp & Dohme. The others have nothing to disclose.

    Financial Support

    This study was supported by grants from the Japanese Ministry of Education, Culture, Sports, Science, and Technology; the Japanese Ministry of Welfare, Health, and Labor; the Japan Society for the Promotion of Science; and the Japan Health Sciences Foundation.

    Electronic supplementary material

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Kawai-Kitahata, F., Asahina, Y., Tanaka, S. et al. Comprehensive analyses of mutations and hepatitis B virus integration in hepatocellular carcinoma with clinicopathological features. J Gastroenterol 51, 473–486 (2016). https://doi.org/10.1007/s00535-015-1126-4

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s00535-015-1126-4

    Keywords

    Search

    Navigation