Skip to main content
NCBI home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1992 Dec;141(6):1271–1277.

Detection of replicative hepatitis C virus sequences in hepatocellular carcinoma.

M A Gerber 1, Y S Shieh 1, K S Shim 1, S N Thung 1, A J Demetris 1, M Schwartz 1, G Akyol 1, S Dash 1
PMCID: PMC1886767  PMID: 1334635

Abstract

Several serologic studies suggest that infection by hepatitis C virus (HCV) may be associated with the development of hepatocellular carcinoma (HCC). Therefore, we examined tumor tissue and/or the surrounding liver of 20 patients for viral sequences by the polymerase chain reaction (PCR). In 12 cases, liver and tumor tissues were separable for extraction. RNA was extracted from frozen tissues and used as a template for reverse transcription followed by double PCR with nested primers for the 5'-untranslated (NT) and nonstructural NS3 regions of HCV. In addition, the tissue extracts were tested by single PCR for X gene and S gene sequences of hepatitis B virus (HBV). NT region sequences of HCV were detected in the available tumor tissue of all anti-HCV-positive patients except for one. Negative (replicative) strands of HCV RNA were found in the same tissues as positive (genomic) strands at almost the same relative amounts, suggesting replication of HCV in the tumor tissue rather than contamination by HCV-positive blood. HBV X and S sequences were demonstrated in two tumors, but were absent from three tumors that were surrounded by liver tissues with HBV X sequences. One patient had nucleic acids of both viruses in tumor tissue. These observations suggest that in addition to HBV, HCV may play a role in the development of hepatocellular carcinoma.

Full text

PDF
1271

Images in this article

1274Figure 1
on p.1274
1275Figure 2
on p.1275

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alter H. J., Purcell R. H., Shih J. W., Melpolder J. C., Houghton M., Choo Q. L., Kuo G. Detection of antibody to hepatitis C virus in prospectively followed transfusion recipients with acute and chronic non-A, non-B hepatitis. N Engl J Med. 1989 Nov 30;321(22):1494–1500. doi: 10.1056/NEJM198911303212202. [DOI] [PubMed] [Google Scholar]
  2. Bruix J., Barrera J. M., Calvet X., Ercilla G., Costa J., Sanchez-Tapias J. M., Ventura M., Vall M., Bruguera M., Bru C. Prevalence of antibodies to hepatitis C virus in Spanish patients with hepatocellular carcinoma and hepatic cirrhosis. Lancet. 1989 Oct 28;2(8670):1004–1006. doi: 10.1016/s0140-6736(89)91015-5. [DOI] [PubMed] [Google Scholar]
  3. Chen M. L., Shieh Y. S., Shim K. S., Gerber M. A. Comparative studies on the detection of hepatitis B virus DNA in frozen and paraffin sections by the polymerase chain reaction. Mod Pathol. 1991 Sep;4(5):555–558. [PubMed] [Google Scholar]
  4. Choo Q. L., Richman K. H., Han J. H., Berger K., Lee C., Dong C., Gallegos C., Coit D., Medina-Selby R., Barr P. J. Genetic organization and diversity of the hepatitis C virus. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2451–2455. doi: 10.1073/pnas.88.6.2451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Colombo M., Kuo G., Choo Q. L., Donato M. F., Del Ninno E., Tommasini M. A., Dioguardi N., Houghton M. Prevalence of antibodies to hepatitis C virus in Italian patients with hepatocellular carcinoma. Lancet. 1989 Oct 28;2(8670):1006–1008. doi: 10.1016/s0140-6736(89)91016-7. [DOI] [PubMed] [Google Scholar]
  6. Cristiano K., Di Bisceglie A. M., Hoofnagle J. H., Feinstone S. M. Hepatitis C viral RNA in serum of patients with chronic non-A, non-B hepatitis: detection by the polymerase chain reaction using multiple primer sets. Hepatology. 1991 Jul;14(1):51–55. doi: 10.1002/hep.1840140109. [DOI] [PubMed] [Google Scholar]
  7. Di Bisceglie A. M., Order S. E., Klein J. L., Waggoner J. G., Sjogren M. H., Kuo G., Houghton M., Choo Q. L., Hoofnagle J. H. The role of chronic viral hepatitis in hepatocellular carcinoma in the United States. Am J Gastroenterol. 1991 Mar;86(3):335–338. [PubMed] [Google Scholar]
  8. Esteban J. I., González A., Hernández J. M., Viladomiu L., Sánchez C., López-Talavera J. C., Lucea D., Martin-Vega C., Vidal X., Esteban R. Evaluation of antibodies to hepatitis C virus in a study of transfusion-associated hepatitis. N Engl J Med. 1990 Oct 18;323(16):1107–1112. doi: 10.1056/NEJM199010183231605. [DOI] [PubMed] [Google Scholar]
  9. Farci P., Alter H. J., Wong D., Miller R. H., Shih J. W., Jett B., Purcell R. H. A long-term study of hepatitis C virus replication in non-A, non-B hepatitis. N Engl J Med. 1991 Jul 11;325(2):98–104. doi: 10.1056/NEJM199107113250205. [DOI] [PubMed] [Google Scholar]
  10. Fong T. L., Shindo M., Feinstone S. M., Hoofnagle J. H., Di Bisceglie A. M. Detection of replicative intermediates of hepatitis C viral RNA in liver and serum of patients with chronic hepatitis C. J Clin Invest. 1991 Sep;88(3):1058–1060. doi: 10.1172/JCI115368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Han J. H., Shyamala V., Richman K. H., Brauer M. J., Irvine B., Urdea M. S., Tekamp-Olson P., Kuo G., Choo Q. L., Houghton M. Characterization of the terminal regions of hepatitis C viral RNA: identification of conserved sequences in the 5' untranslated region and poly(A) tails at the 3' end. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1711–1715. doi: 10.1073/pnas.88.5.1711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hasan F., Jeffers L. J., De Medina M., Reddy K. R., Parker T., Schiff E. R., Houghton M., Choo Q. L., Kuo G. Hepatitis C-associated hepatocellular carcinoma. Hepatology. 1990 Sep;12(3 Pt 1):589–591. doi: 10.1002/hep.1840120323. [DOI] [PubMed] [Google Scholar]
  13. Kaklamani E., Trichopoulos D., Tzonou A., Zavitsanos X., Koumantaki Y., Hatzakis A., Hsieh C. C., Hatziyannis S. Hepatitis B and C viruses and their interaction in the origin of hepatocellular carcinoma. JAMA. 1991 Apr 17;265(15):1974–1976. [PubMed] [Google Scholar]
  14. Kato N., Yokosuka O., Omata M., Hosoda K., Ohto M. Detection of hepatitis C virus ribonucleic acid in the serum by amplification with polymerase chain reaction. J Clin Invest. 1990 Nov;86(5):1764–1767. doi: 10.1172/JCI114903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kiyosawa K., Sodeyama T., Tanaka E., Gibo Y., Yoshizawa K., Nakano Y., Furuta S., Akahane Y., Nishioka K., Purcell R. H. Interrelationship of blood transfusion, non-A, non-B hepatitis and hepatocellular carcinoma: analysis by detection of antibody to hepatitis C virus. Hepatology. 1990 Oct;12(4 Pt 1):671–675. doi: 10.1002/hep.1840120409. [DOI] [PubMed] [Google Scholar]
  16. Lampertico P., Malter J. S., Colombo M., Gerber M. A. Detection of hepatitis B virus DNA in formalin-fixed, paraffin-embedded liver tissue by the polymerase chain reaction. Am J Pathol. 1990 Aug;137(2):253–258. [PMC free article] [PubMed] [Google Scholar]
  17. Levrero M., Tagger A., Balsano C., De Marzio E., Avantaggiati M. L., Natoli G., Diop D., Villa E., Diodati G., Alberti A. Antibodies to hepatitis C virus in patients with hepatocellular carcinoma. J Hepatol. 1991 Jan;12(1):60–63. doi: 10.1016/0168-8278(91)90910-4. [DOI] [PubMed] [Google Scholar]
  18. Liang T. J., Baruch Y., Ben-Porath E., Enat R., Bassan L., Brown N. V., Rimon N., Blum H. E., Wands J. R. Hepatitis B virus infection in patients with idiopathic liver disease. Hepatology. 1991 Jun;13(6):1044–1051. [PubMed] [Google Scholar]
  19. Marcellin P., Martinot-Peignoux M., Loriot M. A., Giostra E., Boyer N., Thiers V., Benhamou J. P. Persistence of hepatitis B virus DNA demonstrated by polymerase chain reaction in serum and liver after loss of HBsAg induced by antiviral therapy. Ann Intern Med. 1990 Feb 1;112(3):227–228. doi: 10.7326/0003-4819-112-3-227. [DOI] [PubMed] [Google Scholar]
  20. Nalpas B., Driss F., Pol S., Hamelin B., Housset C., Brechot C., Berthelot P. Association between HCV and HBV infection in hepatocellular carcinoma and alcoholic liver disease. J Hepatol. 1991 Jan;12(1):70–74. doi: 10.1016/0168-8278(91)90912-u. [DOI] [PubMed] [Google Scholar]
  21. Negro F., Pacchioni D., Shimizu Y., Miller R. H., Bussolati G., Purcell R. H., Bonino F. Detection of intrahepatic replication of hepatitis C virus RNA by in situ hybridization and comparison with histopathology. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2247–2251. doi: 10.1073/pnas.89.6.2247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ogata N., Alter H. J., Miller R. H., Purcell R. H. Nucleotide sequence and mutation rate of the H strain of hepatitis C virus. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3392–3396. doi: 10.1073/pnas.88.8.3392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ohkoshi S., Kojima H., Tawaraya H., Miyajima T., Kamimura T., Asakura H., Satoh A., Hirose S., Hijikata M., Kato N. Prevalence of antibody against non-A, non-B hepatitis virus in Japanese patients with hepatocellular carcinoma. Jpn J Cancer Res. 1990 Jun-Jul;81(6-7):550–553. doi: 10.1111/j.1349-7006.1990.tb02605.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Okamoto H., Okada S., Sugiyama Y., Tanaka T., Sugai Y., Akahane Y., Machida A., Mishiro S., Yoshizawa H., Miyakawa Y. Detection of hepatitis C virus RNA by a two-stage polymerase chain reaction with two pairs of primers deduced from the 5'-noncoding region. Jpn J Exp Med. 1990 Aug;60(4):215–222. [PubMed] [Google Scholar]
  25. Okamoto H., Okada S., Sugiyama Y., Yotsumoto S., Tanaka T., Yoshizawa H., Tsuda F., Miyakawa Y., Mayumi M. The 5'-terminal sequence of the hepatitis C virus genome. Jpn J Exp Med. 1990 Jun;60(3):167–177. [PubMed] [Google Scholar]
  26. Saito I., Miyamura T., Ohbayashi A., Harada H., Katayama T., Kikuchi S., Watanabe Y., Koi S., Onji M., Ohta Y. Hepatitis C virus infection is associated with the development of hepatocellular carcinoma. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6547–6549. doi: 10.1073/pnas.87.17.6547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shieh Y. S., Shim K. S., Lampertico P., Balart L. A., Jeffers L. J., Thung S. N., Regenstein F., Reddy K. R., Farr G., Schiff E. R. Detection of hepatitis C virus sequences in liver tissue by the polymerase chain reaction. Lab Invest. 1991 Oct;65(4):408–411. [PubMed] [Google Scholar]
  28. Shimizu Y. K., Weiner A. J., Rosenblatt J., Wong D. C., Shapiro M., Popkin T., Houghton M., Alter H. J., Purcell R. H. Early events in hepatitis C virus infection of chimpanzees. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6441–6444. doi: 10.1073/pnas.87.16.6441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Takehara T., Hayashi N., Mita E., Hagiwara H., Ueda K., Katayama K., Kasahara A., Fusamoto H., Kamada T. Detection of the minus strand of hepatitis C virus RNA by reverse transcription and polymerase chain reaction: implications for hepatitis C virus replication in infected tissue. Hepatology. 1992 Mar;15(3):387–390. doi: 10.1002/hep.1840150305. [DOI] [PubMed] [Google Scholar]
  30. Weiner A. J., Kuo G., Bradley D. W., Bonino F., Saracco G., Lee C., Rosenblatt J., Choo Q. L., Houghton M. Detection of hepatitis C viral sequences in non-A, non-B hepatitis. Lancet. 1990 Jan 6;335(8680):1–3. doi: 10.1016/0140-6736(90)90134-q. [DOI] [PubMed] [Google Scholar]
  31. Yu M. C., Tong M. J., Coursaget P., Ross R. K., Govindarajan S., Henderson B. E. Prevalence of hepatitis B and C viral markers in black and white patients with hepatocellular carcinoma in the United States. J Natl Cancer Inst. 1990 Jun 20;82(12):1038–1041. doi: 10.1093/jnci/82.12.1038. [DOI] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES