Carcinogenesis

This Article Abstract FREE Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to My Personal Archive Download to citation manager Search for citing articles in: ISI Web of Science (42) Request Permissions Google Scholar Articles by Tsutsumi, M. Articles by Sugimura, T. Search for Related Content PubMed PubMed Citation Articles by Tsutsumi, M. Articles by Sugimura, T. Social Bookmarking          What's this?

Carcinogenesis, Vol. 22, No. 1, 1-3, January 2001
© 2001 Oxford University Press

ACCELERATED PAPER

Increased susceptibility of poly(ADP-ribose) polymerase-1 knockout mice to nitrosamine carcinogenicity

Masahiro Tsutsumi, Mitsuko Masutani^1,,3, Tadashige Nozaki¹, Osamu Kusuoka, Toshifumi Tsujiuchi, Hitoshi Nakagama¹, Hiroshi Suzuki², Yoichi Konishi and Takashi Sugimura¹

Department of Oncological Pathology, Cancer Center, Nara Medical University, 840, Shijo-cho, Kashihara, Nara 634-8521,
¹ Biochemistry Division, National Cancer Center Research Institute, 1-1 Tsukiji 5-chome, Chuo-ku, Tokyo 104-0045 and
² Chugai Pharmaceutical Co. Ltd, 1-135 Komakado, Gotemba, Shizuoka 412-0038, Japan

	Abstract

Top Abstract Introduction References

 
The involvement of poly(ADP-ribose) polymerase-1 (Parp-1), one of the poly(ADP-ribose) polymerase family proteins, in genomic stability, DNA repair and cell death triggered by DNA damage has been well documented. However, the potential role of Parp-1 in carcinogenesis has not been well evaluated. In this study the carcinogenic activity of N-nitrosobis(2-hydroxypropyl)amine (BHP) was studied in Parp-1^–/– mice, generated by disrupting Parp-1 gene exon 1. Parp-1^–/– and Parp-1^+/+ male mice received 0, 250 and 500 p.p.m. BHP in their drinking water for 20 weeks and were then killed. The percentage of animals bearing hemangiomas and hemangiosarcomas in the liver and numbers of tumors per mouse were markedly higher in the Parp-1^–/– groups given 250 or 500 p.p.m. BHP than in their Parp-1^+/+ counterparts. Hemangiosarcomas developed only in Parp-1^–/– mice. In the lung the numbers of adenomas per mouse were increased in Parp-1^–/– mice given BHP at 250 and 500 p.p.m. (P < 0.01) compared with the Parp-1^+/+ case. The results show that susceptibility to BHP is significantly elevated in Parp-1^–/– mice, thus providing direct evidence that Parp-1 is relevant to carcinogenesis.

Abbreviations: BHP, N-nitrosobis(2-hydroxypropyl)amine; Parp, poly(ADP-ribose) polymerase.

	Introduction

Top Abstract Introduction References

 
Poly(ADP-ribose) polymerase (Parp) catalyzes the transfer of ADP-ribose units to various acceptor proteins, including the enzyme itself, resulting in the synthesis of poly(ADP-ribose) from nicotinamide adenine dinucleotide (1,2). Parp-1 is a nuclear enzyme comprising N-terminal DNA-binding, automodification and C-terminal catalytic domains and is activated by DNA strand breaks. The existence of other Parp family proteins (3), including tankyrase (4), PARP-2 (5,6), PARP-3 (5) and VPARP (7), has recently been reported. Parp-1 knockout cells and mice are sensitive to DNA-damaging agents, including alkylating agents (8,9) and -irradiation (8–10), but are resistant to oxidative stress-induced cell death (11–13). Parp-1^–/– mice also show enhanced genomic instability and high frequencies of chromosome aberrations, sister chromatid exchanges and telomere shortening (8,10,14,15). Both gate-keeper and caretaker functions (16) for Parp-1 are implied by this evidence, suggesting an involvement in carcinogenesis. Furthermore, a contribution of change in Parp-1 expression to tumorigenesis is possible since alterations in transcription of genes (17) and cell differentiation during tumor formation (18) have been reported in deficient cells. In the present study increased susceptibility of Parp-1^–/– mice to the carcinogen N-nitrosobis(2-hydroxypropyl)amine (BHP), which induces tumors in multiple organs in rodents (19,20), is documented.

Parp-1^–/– mice with a mixed genetic background of ICR and 129Sv, generated by disrupting Parp-1 exon 1 though insertion of a neomycin resistance gene cassette as described previously (21), were used in this study. Sufficient Parp-1^–/– and Parp-1^+/+animals were obtained by line breeding. A total of 60 male mice aged 6–7 weeks, 30 each of the Parp-1^–/– and Parp-1^+/+ lines, were then divided into six equal groups. Groups 1–3 were Parp-1^–/– mice which received 0, 250 and 500 p.p.m. BHP in their drinking water, respectively, for 20 weeks, while groups 4–6 were Parp-1^+/+ mice given the same doses for the same period. Water containing BHP and a basal diet were available ad libitum and intake was measured once a week. All mice were killed and completely autopsied at 20 weeks. Tumors were processed for routine histological examination. The incidence of liver tumors was determined macroscopically after cutting the whole livers into slices of ~5 mm thickness. The ² and Student's t-tests were performed to compare data for incidences and numbers of liver and lung tumorous lesions in mice, respectively.

The doses of BHP were determined, on the basis of a subacute toxicity study, as those not causing a body weight decrease. The total BHP intakes per Parp-1^–/– mouse given 0, 250 and 500 p.p.m. were 0, 162.5 and 235.7 mg, respectively, and those per Parp-1^+/+ mouse were 0, 179.0 and 239.6 mg, respectively. Final body, liver and lung weights relative to final body weights did not significantly differ among Parp-1^–/– and Parp-1^+/+ mice receiving 0, 250 and 500 p.p.m. BHP. Data for incidences and numbers of tumorous lesions in the liver after BHP administration are summarized in Table I. Neither Parp-1^–/– nor Parp-1^+/+ mice which received 0 p.p.m. BHP showed any spontaneous tumor development in this experimental period (groups 1 and 4, respectively). Hemangiomas developed in Parp-1^+/+ mice that received 500 p.p.m. BHP at a low incidence and number and in Parp-1^–/– mice that received 250 and 500 p.p.m. BHP at a markedly higher incidence and number. In addition, hemangiosarcomas were only observed in Parp-1^–/– mice that received 250 and 500 p.p.m. BHP. Metastases from one such lesion were found in the lung of a Parp-1^–/– mouse that received 500 p.p.m. BHP.

View this table: [in this window] [in a new window]   Table I. Liver tumors in Parp-1–/– and Parp-1+/+ mice given BHP

 
In the lungs of Parp-1^–/– and Parp-1^+/+ mice given BHP alveolar hyperplasias, adenomas and adenocarcinomas were observed. The incidences of mice bearing lung tumors in Parp-1^–/– and Parp-1^+/+ mice at 250 p.p.m. were 5/9 and 5/8, respectively, with no significant difference. However, the incidence in Parp-1^–/– mice was higher than that in Parp-1^+/+ mice with 500 p.p.m., at 8/9 and 4/8, respectively. Furthermore, compared with the Parp-1^+/+ case, the numbers of lung adenomas per mouse in Parp-1^–/– mice were higher at both 250 (3.2 ± 3.8 and 1.4 ± 1.7, respectively) and 500 p.p.m. (3.0 ± 2.5 and 0.9 ± 1.4, respectively; P < 0.01). These results for liver and lung lesions demonstrated elevated susceptibility of Parp-1^–/– mice to BHP carcinogenicity.

BHP is reported to be an environmental contaminant of commercial samples of diisopropanolamine and triisopropanolamine (22) which is metabolized through oxidation by cytochrome P450 enzymes primarily in the liver, finally generating methylating or hydroxypropylating species (23,24). Methylated bases produced by BHP, including O⁶- and N⁷-methylguanine, can be repaired by alkylguanine alkyltransferases or base excision repair. Hydroxypropylated guanines such as O⁶- and N⁷-hydroxypropylguanine might be repaired through nucleotide excision repair. The elevated susceptibility to carcinogenicity induced by BHP in Parp-1^–/– mice compared with Parp-1^+/+ mice may be due to a deficiency in base excision repair (25) or other DNA repair pathways. The resistance of Parp-1^–/– cells to cell death after extensive DNA damage (13) could also be related to a higher sensitivity to the carcinogen. Not only initiation of carcinogenesis through gene mutation but also tumor progression may be enhanced in the Parp-1-deficient state, as hemangiosarcomas only developed from hemangiomas in Parp-1^–/– animals. This malignant progression could have been caused by dysfunction of Parp-1 in transcriptional regulation of genes involved in cell differentiation or maintenance of genomic integrity.

Increased incidences of spontaneous tumors have not been found in any organs of Parp-1^–/– mice (8,10,21) other than a reported higher level of spontaneous T cell lymphomas in Parp-1^–/– SCID compared with Parp-1^+/+ SCID mice (26). Furthermore, although alterations in Parp-1 expression in cancer cells have been described (27), inactivation or genetic alteration of the Parp-1 gene in neoplasia has not been reported so far. The present study clearly indicates that dysfunction of Parp-1 could be considered as one of the risk factors for carcinogenesis. Thus Parp-1^–/– mice should provide a useful animal model for the sensitive detection of carcinogenicity of genotoxic agents.

	Notes

 
³ To whom correspondence should be addressed Email: mmasutan{at}gan2.ncc.go.jp

	Acknowledgments

 
This work was supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Health and Welfare of Japan and by a Grant-in Aid for the Second Term Comprehensive 10-Year Strategy for Cancer Control, Cancer Prevention, from the Ministry of Health and Welfare of Japan.

	References

Top Abstract Introduction References

 

1. Sugimura,T. (>1973) Poly(adenosine diphosphate ribose). Prog. Nucleic Acid Res. Mol. Biol., B, 127–151.
2. Althaus,F.R. and Richter,C. (eds) (1987) ADP-ribosylation of Proteins. Springer-Verlag, Berlin, Germany.
3. Shieh,W.M., Ame,J.C., Wilson,M.V., Wang,Z.Q., Koh,D.W., Jacobson, M.K. and Jacobson,E.L. (1998) Poly(ADP-ribose) polymerase null mouse cells synthesize ADP-ribose polymers. J. Biol. Chem., 273, 30069–30072.[Abstract/Free Full Text]
4. Smith,S., Giriat I., Schmitt,A. and de Lange,T. (1998) Tankyrase, a poly(ADP-ribose) polymerase at human telomeres. Science, 282, 1484–1487.[Abstract/Free Full Text]
5. Johansson,M. (1999) A human poly(ADP-ribose) polymerase gene family (ADPRTL): cDNA cloning of two novel poly(ADP-ribose) homologues. Genomics, 57, 442–445.[Web of Science][Medline]
6. Berghammer,H., Ebner,M., Marksteiner,R. and Aur,B. (1999) pADPRT-2: a novel mammalian polymerizing(ADP-ribosyl)transferase gene related to truncated pADPRT homologues in plants and Caenorhabditis elegans. FEBS Lett., 449, 259–263.[Web of Science][Medline]
7. Kickhoefer,V.A., Siva,A.C., Kedersha,N.L., Inman,E.M., Ruland,C., Streuli,M. and Rome,L.H. (1999) The 193-kD vault protein, VPARP, is a novel poly(ADP-ribose) polymerase. J. Cell Biol., 146, 917–928.[Abstract/Free Full Text]
8. de Murcia,J.M., Niedergang,C., Trucco,C., Ricoul,M., Dutrillaux,B., Mark,M., Oliver,F.J., Masson,M., Dierich,A., LeMeur,M., Walztinger,C., Chambon,P. and de Murcia,G. (1997) Requirement of poly(ADP-ribose) polymerase in recovery from DNA damage in mice and in cells. Proc. Natl Acad. Sci. USA, 94, 7303–7307.[Abstract/Free Full Text]
9. Masutani,M., Nozaki,T., Nakamoto,K., Nakagama,H., Suzuki,H., Kusuoka,O., Tsutsumi,M. and Sugimura,T. (2000) The response of Parp knockout mice against DNA damaging agents. Mutat. Res., 462, 159–166.[Web of Science][Medline]
10. Wang,Z.Q., Stingl,L., Morrison,C., Jantsch,M., Los,M., Schulze-Osthoff,K. and Wagner,E.F. (1997) PARP-1 is important for genomic stability but dispensable in apoptosis. Genes Dev., 11, 2347–2358.[Abstract/Free Full Text]
11. Heller,B., Wang,Z.Q., Wagner,E.F., Radons,J., Bürkle,A., Fehsel,K., Burkart,V. and Kolb,H. (1995) Inactivation of the poly(ADP-ribose) polymerase gene affects oxygen radical and nitric oxide toxicity in islet cells. J. Biol. Chem., 270, 11176–11180.[Abstract/Free Full Text]
12. Eliasson,M.J., Sampei,K., Mandir,A.S., Hurn,P.D., Traystman,R.J., Bao,J., Pieper,A., Wang,Z.Q., Dawson,T.M., Snyder,S.H. and Dawson,V.L. (1997) Poly(ADP-ribose) polymerase gene disruption renders mice resistant to cerebral ischemia. Nature Med., 3, 1089–1095.[Web of Science][Medline]
13. Ha,H.C. and Snyder,S.H. (1999) Poly(ADP-ribose) polymerase is a mediator of necrotic cell death by ATP depletion. Proc. Natl Acad. Sci. USA, 96, 13978–13982.[Abstract/Free Full Text]
14. Fagagna,F.D., Hande,M.P., Tong,W.M., Lansdorp,P.M., Wang,Z.Q. and Jackson,S.P. (1999) Functions of poly(ADP-ribose) polymerase in controlling telomere length and chromosomal stability. Nature Genet., 23, 76–80.[Web of Science][Medline]
15. Simbulan-Rosenthal,C.M., Haddad,B.R., Rosenthal,D.S., Weaver,Z., Coleman,A., Luo,R., Young,H.M., Wang,Z.Q., Ried,T. and Smulson,M.E. (1999) Chromosomal aberrations in PARP-1 (–/–) mice: Genome stabilization in immortalized cells by reintroduction of poly(ADP-ribose) polymerase cDNA. Proc. Natl Acad. Sci. USA, 96, 13191–13196.[Abstract/Free Full Text]
16. Kinzler,K.W. and Vogelstein,B. (1997) Gatekeepers and caretakers. Nature, 386, 761–763.[Medline]
17. Oliver,F.J., Menissier-de Murcia,J., Nacci,C., Decker,P., Andriantsitohaina,R., Muller,S., de la Rubia,G., Stoclet,J.C. and de Murcia,G. (1999) Resistance to endotoxic shock as a consequence of defective NF-B activation in poly(ADP-ribose) polymerase-1 deficient mice. EMBO J., 18, 4446–4454.[Web of Science][Medline]
18. Nozaki,T., Masutani,M., Watanabe,M., Ochiya,T., Hasegawa,F., Nakagama,H., Suzuki,H. and Sugimura,T. (1999) Syncytiotrophoblastic giant cells in teratocarcinoma-like tumors derived from Parp-disrupted mouse embryonic stem cells. Proc. Natl Acad. Sci. USA, 96, 13345–13350.[Abstract/Free Full Text]
19. Konishi,Y., Kondo,H., Inui,S., Denda,A., Ikeda,T. and Kojima,K. (1977) Carcinogenic effect of N-bis(2-hydroxypropyl)-nitrosamine by oral administration in mice. Cancer Lett., 3, 255–257.
20. Konishi,Y., Ikeda,T., Yoshimura,H., Kawabata,A. and Mikami,R. (1979) Carcinogenic effect of N-bis(2-hydroxypropyl)nitrosamine by a single administration in rats. Cancer Lett., 6, 115–119.[Web of Science][Medline]
21. Masutani,M., Suzuki,H., Kamada,N., Watanabe,M., Ueda,O., Nozaki,T., Jishage,K., Watanabe,T., Sugimoto,T., Nakagama,H., Ochiya,T. and Sugimura,T. (1999) Poly(ADP-ribose) polymerase gene disruption conferred mice resistant to streptozotocin-induced diabetes. Proc. Natl Acad. Sci. USA, 96, 2301–2304.[Abstract/Free Full Text]
22. Issenberg,P., Conrad,E.E., Nielsen,J.W., Klein,D.A. and Miller,S.E. (1984) Determination of N-Nitrosobis(2-hydroxypropyl)amine in Environmental Samples, IARC Scientific Publications no. 57. IARC, Lyon, pp. 43–50.
23. Mori,Y., Takahashi,H., Yamazaki,H., Toyoshi,K., Makino,T., Yokose,Y. and Konishi,Y. (1984) Distribution, metabolism and excretion of N-nitrosobis(2-hydroxypropyl)amine in Wistar rats. Carcinogenesis, 5, 1443–1447.[Abstract/Free Full Text]
24. Kokkinakis,D.M. (1992) Alkylation of rodent tissue DNA induced by N-nitrosobis(2-hydroxypropyl)amine. Carcinogenesis, 13, 759–765.
25. Trucco,C., Oliver,F.J., de Murcia,G. and de Murcia,J.M. (1998) DNA repair defect in poly(ADP-ribose) polymerase-deficient cell lines. Nucleic Acids Res., 11, 2644–2649.
26. Morrison,C., Smith,G.C., Stingl,L., Jackson,S.P., Wang,Z.Q. and Wagner,E.F. (1997) Genetic interaction between PARP-1 and DNA-PK in V(D)J recombination and tumorigenesis. Nature Genet., 17, 479–482.[Web of Science][Medline]
27. Bieche,I, de Murcia,G. and Lidereau,R. (1996) Poly(ADP-ribose), polymerase gene expression status and genomic instability in human breast cancer. Clin. Cancer Res., 2, 1163–1167.[Abstract]

Received June 29, 2000; revised October 4, 2000; accepted October 6, 2000.

CiteULike    Connotea    Del.icio.us    What's this?

This article has been cited by other articles:

				S. Landi, F. Gemignani, F. Canzian, V. Gaborieau, R. Barale, D. Landi, N. Szeszenia-Dabrowska, D. Zaridze, J. Lissowska, P. Rudnai, et al. DNA Repair and Cell Cycle Control Genes and the Risk of Young-Onset Lung Cancer. Cancer Res., November 15, 2006; 66(22): 11062 - 11069. [Abstract] [Full Text] [PDF]

				S. Raval-Fernandes, V. A. Kickhoefer, C. Kitchen, and L. H. Rome Increased Susceptibility of Vault Poly(ADP-Ribose) Polymerase-Deficient Mice to Carcinogen-Induced Tumorigenesis Cancer Res., October 1, 2005; 65(19): 8846 - 8852. [Abstract] [Full Text] [PDF]

				M. Y. Kim, T. Zhang, and W. L. Kraus Poly(ADP-ribosyl)ation by PARP-1: `PAR-laying' NAD+ into a nuclear signal Genes & Dev., September 1, 2005; 19(17): 1951 - 1967. [Abstract] [Full Text] [PDF]

				M. Shiokawa, M. Masutani, H. Fujihara, K. Ueki, R. Nishikawa, T. Sugimura, H. Kubo, and H. Nakagama Genetic Alteration of Poly(ADP-ribose) Polymerase-1 in Human Germ Cell Tumors Jpn. J. Clin. Oncol., February 1, 2005; 35(2): 97 - 102. [Abstract] [Full Text] [PDF]

				M. Lebel, J. Lavoie, I. Gaudreault, M. Bronsard, and R. Drouin Genetic Cooperation between the Werner Syndrome Protein and Poly(ADP-Ribose) Polymerase-1 in Preventing Chromatid Breaks, Complex Chromosomal Rearrangements, and Cancer in Mice Am. J. Pathol., May 1, 2003; 162(5): 1559 - 1569. [Abstract] [Full Text] [PDF]

				W.-M. Tong, H. Ohgaki, H. Huang, C. Granier, P. Kleihues, and Z.-Q. Wang Null Mutation of DNA Strand Break-Binding Molecule Poly(ADP-ribose) Polymerase Causes Medulloblastomas in p53-/- Mice Am. J. Pathol., January 1, 2003; 162(1): 343 - 352. [Abstract] [Full Text] [PDF]

				W.-M. Tong, U. Cortes, M. P. Hande, H. Ohgaki, L. R. Cavalli, P. M. Lansdorp, B. R. Haddad, and Z.-Q. Wang Synergistic Role of Ku80 and Poly(ADP-ribose) Polymerase in Suppressing Chromosomal Aberrations and Liver Cancer Formation Cancer Res., December 1, 2002; 62(23): 6990 - 6996. [Abstract] [Full Text] [PDF]

				M. Masutani and M. Miwa Poly(ADP-ribose) Polymerase and Cancer: In Relation to the Lectures Presented by Dr Gilbert de Murcia Jpn. J. Clin. Oncol., November 1, 2002; 32(11): 483 - 487. [Full Text] [PDF]

				L. Virag and C. Szabo The Therapeutic Potential of Poly(ADP-Ribose) Polymerase Inhibitors Pharmacol. Rev., September 1, 2002; 54(3): 375 - 429. [Abstract] [Full Text] [PDF]

This Article Abstract FREE Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to My Personal Archive Download to citation manager Search for citing articles in: ISI Web of Science (42) Request Permissions Google Scholar Articles by Tsutsumi, M. Articles by Sugimura, T. Search for Related Content PubMed PubMed Citation Articles by Tsutsumi, M. Articles by Sugimura, T. Social Bookmarking          What's this?

Online ISSN 1460-2180 - Print ISSN 0143-3334

Copyright © 2009 Oxford University Press

Oxford Journals Oxford University Press

• Site Map
• Privacy Policy
• Frequently Asked Questions

Other Oxford University Press sites: Oxford University Press Oxford Journals China Oxford Journals Japan American National Biography Booksellers' Information Service Children's Fiction and Poetry Children's Reference Corporate & Special Sales Dictionaries Dictionary of National Biography Digital Reference English Language Teaching Higher Education Textbooks Humanities International Education Unit Law Medicine Music Online Products Oxford English Dictionary Reference Rights and Permissions Science School Books Social Sciences Very Short Introductions World's Classics