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Proc Natl Acad Sci U S A. Dec 20, 1994; 91(26): 12808–12812.
PMCID: PMC45529

Extensive oxidative DNA damage in hepatocytes of transgenic mice with chronic active hepatitis destined to develop hepatocellular carcinoma.

Abstract

A transgenic mouse strain that expresses the hepatitis B virus (HBV) large envelope protein in the liver was used to determine the extent of oxidative DNA damage that occurs during chronic HBV infection. This mouse strain develops a chronic necroinflammatory liver disease that mimics the inflammation, cellular hyperplasia, and increased risk for cancer that is evident in human chronic active hepatitis. When perfused in situ with nitroblue tetrazolium, an indicator for superoxide formation, the liver of transgenic mice displayed intense formazan deposition in Kupffer cells, indicating oxygen radical production, and S-phase hepatocytes were commonly seen adjacent to the stained Kupffer cells. Similar changes were not observed in nontransgenic control livers. To determine whether these events were associated with oxidative DNA damage, genomic DNA from the livers of transgenic mice and nontransgenic controls was isolated and examined for 8-oxo-2'-deoxyguanosine, an oxidatively modified adduct of deoxyguanosine. Results showed a significant, sustained accumulation in steady-state 8-oxo-2'-deoxyguanosine that started early in life exclusively in the transgenic mice and increased progressively with advancing disease. The most pronounced increase occurred in livers exhibiting microscopic nodular hyperplasia, adenomas, and hepatocellular carcinoma. Thus, HBV transgenic mice with chronic active hepatitis display greatly increased hepatic oxidative DNA damage. Moreover, the DNA damage occurs in the presence of heightened hepatocellular proliferation, increasing the probability of fixation of the attendant genetic and chromosomal abnormalities and the development of hepatocellular carcinoma.

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  • Ames BN, Shigenaga MK, Hagen TM. Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):7915–7922. [PMC free article] [PubMed]
  • Tiollais P, Pourcel C, Dejean A. The hepatitis B virus. Nature. 1985 Oct 10;317(6037):489–495. [PubMed]
  • Di Bisceglie AM, Rustgi VK, Hoofnagle JH, Dusheiko GM, Lotze MT. NIH conference. Hepatocellular carcinoma. Ann Intern Med. 1988 Mar;108(3):390–401. [PubMed]
  • Kew MC, Popper H. Relationship between hepatocellular carcinoma and cirrhosis. Semin Liver Dis. 1984 May;4(2):136–146. [PubMed]
  • Sakuma K, Saitoh N, Kasai M, Jitsukawa H, Yoshino I, Yamaguchi M, Nobutomo K, Yamumi M, Tsuda F, Komazawa T, et al. Relative risks of death due to liver disease among Japanese male adults having various statuses for hepatitis B s and e antigen/antibody in serum: a prospective study. Hepatology. 1988 Nov-Dec;8(6):1642–1646. [PubMed]
  • Tiollais P, Buendia MA. Hepatitis B virus. Sci Am. 1991 Apr;264(4):116–123. [PubMed]
  • De Mitri MS, Pisi E, Bréchot C, Paterlini P. Low frequency of allelic loss in the cyclin A gene in human hepatocellular carcinomas: a study based on PCR. Liver. 1993 Oct;13(5):259–261. [PubMed]
  • Wang J, Chenivesse X, Henglein B, Bréchot C. Hepatitis B virus integration in a cyclin A gene in a hepatocellular carcinoma. Nature. 1990 Feb 8;343(6258):555–557. [PubMed]
  • Himeno Y, Fukuda Y, Hatanaka M, Imura H. Expression of oncogenes in human liver disease. Liver. 1988 Aug;8(4):208–212. [PubMed]
  • Pasquinelli C, Bhavani K, Chisari FV. Multiple oncogenes and tumor suppressor genes are structurally and functionally intact during hepatocarcinogenesis in hepatitis B virus transgenic mice. Cancer Res. 1992 May 15;52(10):2823–2829. [PubMed]
  • Hsu IC, Tokiwa T, Bennett W, Metcalf RA, Welsh JA, Sun T, Harris CC. p53 gene mutation and integrated hepatitis B viral DNA sequences in human liver cancer cell lines. Carcinogenesis. 1993 May;14(5):987–992. [PubMed]
  • Thomas HC. The hepatitis B virus and the host response. J Hepatol. 1990;11 (Suppl 1):S83–S89. [PubMed]
  • Weitzman SA, Gordon LI. Inflammation and cancer: role of phagocyte-generated oxidants in carcinogenesis. Blood. 1990 Aug 15;76(4):655–663. [PubMed]
  • Columbano A, Rajalakshmi S, Sarma DS. Requirement of cell proliferation for the initiation of liver carcinogenesis as assayed by three different procedures. Cancer Res. 1981 Jun;41(6):2079–2083. [PubMed]
  • Lewis JG, Adams DO. Inflammation, oxidative DNA damage, and carcinogenesis. Environ Health Perspect. 1987 Dec;76:19–27. [PMC free article] [PubMed]
  • Chisari FV, Pinkert CA, Milich DR, Filippi P, McLachlan A, Palmiter RD, Brinster RL. A transgenic mouse model of the chronic hepatitis B surface antigen carrier state. Science. 1985 Dec 6;230(4730):1157–1160. [PubMed]
  • Chisari FV, Klopchin K, Moriyama T, Pasquinelli C, Dunsford HA, Sell S, Pinkert CA, Brinster RL, Palmiter RD. Molecular pathogenesis of hepatocellular carcinoma in hepatitis B virus transgenic mice. Cell. 1989 Dec 22;59(6):1145–1156. [PubMed]
  • Chisari FV, Filippi P, McLachlan A, Milich DR, Riggs M, Lee S, Palmiter RD, Pinkert CA, Brinster RL. Expression of hepatitis B virus large envelope polypeptide inhibits hepatitis B surface antigen secretion in transgenic mice. J Virol. 1986 Dec;60(3):880–887. [PMC free article] [PubMed]
  • Chisari FV, Filippi P, Buras J, McLachlan A, Popper H, Pinkert CA, Palmiter RD, Brinster RL. Structural and pathological effects of synthesis of hepatitis B virus large envelope polypeptide in transgenic mice. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6909–6913. [PMC free article] [PubMed]
  • Dunsford HA, Sell S, Chisari FV. Hepatocarcinogenesis due to chronic liver cell injury in hepatitis B virus transgenic mice. Cancer Res. 1990 Jun 1;50(11):3400–3407. [PubMed]
  • Blobel G, Potter VR. Nuclei from rat liver: isolation method that combines purity with high yield. Science. 1966 Dec 30;154(3757):1662–1665. [PubMed]
  • Shigenaga MK, Aboujaoude EN, Chen Q, Ames BN. Assays of oxidative DNA damage biomarkers 8-oxo-2'-deoxyguanosine and 8-oxoguanine in nuclear DNA and biological fluids by high-performance liquid chromatography with electrochemical detection. Methods Enzymol. 1994;234:16–33. [PubMed]
  • Fry JR, Jones CA, Wiebkin P, Bellemann P, Bridges JW. The enzymic isolation of adult rat hepatocytes in a functional and viable state. Anal Biochem. 1976 Apr;71(2):341–350. [PubMed]
  • Lewis JG, Adams DO. Induction of 5,6-ring-saturated thymine bases in NIH-3T3 cells by phorbol ester-stimulated macrophages: role of reactive oxygen intermediates. Cancer Res. 1985 Mar;45(3):1270–1275. [PubMed]
  • Thoolen B. BrdUrd labeling of S-phase cells in testes and small intestine of mice, using microwave irradiation for immunogold-silver staining: an immunocytochemical study. J Histochem Cytochem. 1990 Feb;38(2):267–273. [PubMed]
  • Jezequel AM, Paolucci F, Benedetti A, Mancini R, Orlandi F. Enumeration of S-phase cells in normal rat liver by immunohistochemistry using bromodeoxyuridine-antibromodeoxyuridine system. Dig Dis Sci. 1991 Apr;36(4):482–484. [PubMed]
  • Fraga CG, Shigenaga MK, Park JW, Degan P, Ames BN. Oxidative damage to DNA during aging: 8-hydroxy-2'-deoxyguanosine in rat organ DNA and urine. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4533–4537. [PMC free article] [PubMed]
  • Kasai H, Nishimura S. Hydroxylation of deoxyguanosine at the C-8 position by ascorbic acid and other reducing agents. Nucleic Acids Res. 1984 Feb 24;12(4):2137–2145. [PMC free article] [PubMed]
  • Kasai H, Crain PF, Kuchino Y, Nishimura S, Ootsuyama A, Tanooka H. Formation of 8-hydroxyguanine moiety in cellular DNA by agents producing oxygen radicals and evidence for its repair. Carcinogenesis. 1986 Nov;7(11):1849–1851. [PubMed]
  • Kasai H, Nishimura S, Kurokawa Y, Hayashi Y. Oral administration of the renal carcinogen, potassium bromate, specifically produces 8-hydroxydeoxyguanosine in rat target organ DNA. Carcinogenesis. 1987 Dec;8(12):1959–1961. [PubMed]
  • Kuchino Y, Mori F, Kasai H, Inoue H, Iwai S, Miura K, Ohtsuka E, Nishimura S. Misreading of DNA templates containing 8-hydroxydeoxyguanosine at the modified base and at adjacent residues. Nature. 1987 May 7;327(6117):77–79. [PubMed]
  • Kasai H, Chung MH, Yamamoto F, Ohtsuka E, Laval J, Grollman AP, Nishimura S. Formation, inhibition of formation, and repair of oxidative 8-hydroxyguanine DNA damage. Basic Life Sci. 1993;61:257–262. [PubMed]
  • Shimoda R, Nagashima M, Sakamoto M, Yamaguchi N, Hirohashi S, Yokota J, Kasai H. Increased formation of oxidative DNA damage, 8-hydroxydeoxyguanosine, in human livers with chronic hepatitis. Cancer Res. 1994 Jun 15;54(12):3171–3172. [PubMed]
  • Colombo M, Kuo G, Choo QL, Donato MF, Del Ninno E, Tommasini MA, 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. [PubMed]
  • Trichopoulos D, Day NE, Tzonou A, Hadziyannis S, Kaklamani E, Sparos L, Muñoz N, Hatzakis A. Delta agent and the etiology of hepatocellular carcinoma. Int J Cancer. 1987 Mar 15;39(3):283–286. [PubMed]
  • Kurathong S, Lerdverasirikul P, Wongpaitoon V, Pramoolsinsap C, Kanjanapitak A, Varavithya W, Phuapradit P, Bunyaratvej S, Upatham ES, Brockelman WY. Opisthorchis viverrini infection and cholangiocarcinoma. A prospective, case-controlled study. Gastroenterology. 1985 Jul;89(1):151–156. [PubMed]
  • Parkin DM, Srivatanakul P, Khlat M, Chenvidhya D, Chotiwan P, Insiripong S, L'Abbé KA, Wild CP. Liver cancer in Thailand. I. A case-control study of cholangiocarcinoma. Int J Cancer. 1991 May 30;48(3):323–328. [PubMed]
  • Shindo K. Significance of Schistosomiasis japonica in the development of cancer of the large intestine: report of a case and review of the literature. Dis Colon Rectum. 1976 Jul-Aug;19(5):460–469. [PubMed]
  • Rosin MP, Saad el Din Zaki S, Ward AJ, Anwar WA. Involvement of inflammatory reactions and elevated cell proliferation in the development of bladder cancer in schistosomiasis patients. Mutat Res. 1994 Mar 1;305(2):283–292. [PubMed]
  • Purtilo DT. Clonorchiasis and hepatic neoplasms. Trop Geogr Med. 1976 Mar;28(1):21–27. [PubMed]
  • Parsonnet J. Helicobacter pylori and gastric cancer. Gastroenterol Clin North Am. 1993 Mar;22(1):89–104. [PubMed]
  • Daniel LN, Mao Y, Saffiotti U. Oxidative DNA damage by crystalline silica. Free Radic Biol Med. 1993 May;14(5):463–472. [PubMed]
  • Rom WN, Travis WD, Brody AR. Cellular and molecular basis of the asbestos-related diseases. Am Rev Respir Dis. 1991 Feb;143(2):408–422. [PubMed]

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