Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions


DNA damage checkpoint genes, such as p53, are frequently mutated in human cancer, but the selective pressure for their inactivation remains elusive1,2,3. We analysed a panel of human lung hyperplasias, all of which retained wild-type p53 genes and had no signs of gross chromosomal instability, and found signs of a DNA damage response, including histone H2AX and Chk2 phosphorylation, p53 accumulation, focal staining of p53 binding protein 1 (53BP1) and apoptosis. Progression to carcinoma was associated with p53 or 53BP1 inactivation and decreased apoptosis. A DNA damage response was also observed in dysplastic nevi and in human skin xenografts, in which hyperplasia was induced by overexpression of growth factors. Both lung and experimentally-induced skin hyperplasias showed allelic imbalance at loci that are prone to DNA double-strand break formation when DNA replication is compromised (common fragile sites). We propose that, from its earliest stages, cancer development is associated with DNA replication stress, which leads to DNA double-strand breaks, genomic instability and selective pressure for p53 mutations.

This is a preview of subscription content

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Activation of the DNA double-strand break (DSB) checkpoint pathway in human preneoplastic and neoplastic lesions.
Figure 2: Summary of DNA double-strand break responses in human normal tissues, preneoplastic lesions and neoplastic lesions.
Figure 3: Activation of the DNA DSB checkpoint pathway in experimentally induced hyperplasias.
Figure 4: Allelic imbalance at common fragile sites in early human cancer lesions and model for activation of the DNA DSB checkpoint in cancer.


  1. 1

    Hollstein, M., Sidransky, D., Vogelstein, B. & Harris, C. C. p53 mutations in human cancers. Science 253, 49–53 (1991)

    ADS  CAS  Article  Google Scholar 

  2. 2

    Kastan, M. B. & Bartek, J. Cell-cycle checkpoints and cancer. Nature 432, 316–323 (2004)

    ADS  CAS  Article  Google Scholar 

  3. 3

    Halazonetis, T. D. Constitutively active DNA damage checkpoint pathways as the driving force for the high frequency of p53 mutations in human cancer. DNA Repair (Amst.) 3, 1057–1062 (2004)

    CAS  Article  Google Scholar 

  4. 4

    Takai, H., Smogorzewska, A. & de Lange, T. DNA damage foci at dysfunctional telomeres. Curr. Biol. 13, 1549–1556 (2003)

    CAS  Article  Google Scholar 

  5. 5

    d'Adda di Fagagna, F. et al. A DNA damage checkpoint response in telomere-initiated senescence. Nature 426, 194–198 (2003)

    ADS  CAS  Article  Google Scholar 

  6. 6

    Graeber, T. G. et al. Hypoxia-mediated selection of cells with diminished apoptotic potential in solid tumours. Nature 379, 88–91 (1996)

    ADS  CAS  Article  Google Scholar 

  7. 7

    Gorgoulis, V. G. et al. Alterations of the p16-pRb pathway and the chromosome locus 9p21–22 in non-small-cell lung carcinomas: relationship with p53 and MDM2 protein expression. Am. J. Pathol. 153, 1749–1765 (1998)

    CAS  Article  Google Scholar 

  8. 8

    Karakaidos, P. et al. Overexpression of the replication licensing regulators hCdt1 and hCdc6 characterizes a subset of non-small-cell lung carcinomas: synergistic effect with mutant p53 on tumor growth and chromosomal instability—evidence of E2F-1 transcriptional control over hCdt1. Am. J. Pathol. 165, 1351–1365 (2004)

    CAS  Article  Google Scholar 

  9. 9

    Kastan, M. B., Onyekwere, O., Sidransky, D., Vogelstein, B. & Craig, R. W. Participation of p53 protein in the cellular response to DNA damage. Cancer Res. 51, 6304–6311 (1991)

    CAS  PubMed  Google Scholar 

  10. 10

    Rogakou, E. P., Pilch, D. R., Orr, A. H., Ivanova, V. S. & Bonner, W. M. DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139. J. Biol. Chem. 273, 5858–5868 (1998)

    CAS  Article  Google Scholar 

  11. 11

    Bartek, J. & Lukas, J. Chk1 and Chk2 kinases in checkpoint control and cancer. Cancer Cell 3, 421–429 (2003)

    CAS  Article  Google Scholar 

  12. 12

    Schultz, L. B., Chehab, N. H., Malikzay, A. & Halazonetis, T. D. p53 binding protein 1 (53BP1) is an early participant in the cellular response to DNA double-strand breaks. J. Cell Biol. 151, 1381–1390 (2000)

    CAS  Article  Google Scholar 

  13. 13

    Mochan, T. A., Venere, M., DiTullio, R. A. Jr & Halazonetis, T. D. 53BP1 and NFBD1/MDC1-Nbs1 function in parallel interacting pathways activating ataxia-telangiectasia mutated (ATM) in response to DNA damage. Cancer Res. 63, 8586–8591 (2003)

    CAS  PubMed  Google Scholar 

  14. 14

    Huyen, Y. et al. Methylated lysine 79 of histone H3 targets 53BP1 to DNA double-strand breaks. Nature 432, 406–411 (2004)

    ADS  CAS  Article  Google Scholar 

  15. 15

    DiTullio, R. A. Jr et al. 53BP1 functions in an ATM-dependent checkpoint pathway that is constitutively activated in human cancer. Nature Cell Biol. 4, 998–1002 (2002)

    CAS  Article  Google Scholar 

  16. 16

    Bartkova, J. et al. DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. Nature doi:10.1038/nature03482 (this issue)

  17. 17

    Berking, C. et al. Induction of melanoma phenotypes in human skin by growth factors and ultraviolet B. Cancer Res. 64, 807–811 (2004)

    CAS  Article  Google Scholar 

  18. 18

    Spruck, C. H., Won, K. A. & Reed, S. I. Deregulated cyclin E induces chromosome instability. Nature 401, 297–300 (1999)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Lengronne, A. & Schwob, E. The yeast CDK inhibitor Sic1 prevents genomic instability by promoting replication origin licensing in late G1. Mol. Cell 9, 1067–1078 (2002)

    CAS  Article  Google Scholar 

  20. 20

    Tanaka, S. & Diffley, J. F. Deregulated G1-cyclin expression induces genomic instability by preventing efficient pre-RC formation. Genes Dev. 16, 2639–2649 (2002)

    CAS  Article  Google Scholar 

  21. 21

    Vaziri, C. et al. A p53-dependent checkpoint pathway prevents rereplication. Mol. Cell 11, 997–1008 (2003)

    CAS  Article  Google Scholar 

  22. 22

    Ekholm-Reed, S. et al. Deregulation of cyclin E in human cells interferes with prereplication complex assembly. J. Cell Biol. 165, 789–800 (2004)

    CAS  Article  Google Scholar 

  23. 23

    Tibbetts, R. S. et al. Functional interactions between BRCA1 and the checkpoint kinase ATR during genotoxic stress. Genes Dev. 14, 2989–3002 (2000)

    CAS  Article  Google Scholar 

  24. 24

    Cortez, D., Guntuku, S., Qin, J. & Elledge, S. J. ATR and ATRIP: partners in checkpoint signaling. Science 294, 1713–1716 (2001)

    ADS  CAS  Article  Google Scholar 

  25. 25

    MacPail, S. H., Banath, J. P., Yu, Y., Chu, E. & Olive, P. L. Cell cycle-dependent expression of phosphorylated histone H2AX: reduced expression in unirradiated but not X-irradiated G1-irradiated G1-phase cells. Radiat. Res. 159, 759–767 (2003)

    ADS  Article  Google Scholar 

  26. 26

    Arlt, M. F., Casper, A. M. & Glover, T. W. Common fragile sites. Cytogenet. Genome Res. 100, 92–100 (2003)

    CAS  Article  Google Scholar 

  27. 27

    Casper, A. M., Nghiem, P., Arlt, M. F. & Glover, T. W. ATR regulates fragile site stability. Cell 111, 779–789 (2002)

    CAS  Article  Google Scholar 

  28. 28

    Mao, L. et al. Frequent microsatellite alterations at chromosomes 9p21 and 3p14 in oral premalignant lesions and their value in cancer risk assessment. Nature Med. 2, 682–685 (1996)

    CAS  Article  Google Scholar 

  29. 29

    Wistuba, I. I. et al. High resolution chromosome 3p allelotyping of human lung cancer and preneoplastic/preinvasive bronchial epithelium reveals multiple, discontinuous sites of 3p allele loss and three regions of frequent breakpoints. Cancer Res. 60, 1949–1960 (2000)

    CAS  PubMed  Google Scholar 

  30. 30

    Maitra, A. et al. High-resolution chromosome 3p allelotyping of breast carcinomas and precursor lesions demonstrates frequent loss of heterozygosity and a discontinuous pattern of allele loss. Am. J. Pathol. 159, 119–130 (2001)

    CAS  Article  Google Scholar 

Download references


The authors thank R. Kaufman, L. Kaklamanis, M. Arnaouti, P. Foukas, K. Ryan and V. Kostaki for support, reagents and tissue samples. This work was supported by grants to T.D.H. from the National Cancer Institute and to T.L. from the Roy Castle Lung Foundation, UK. M.V. was supported by a Radiation training grant from the NIH.

Author information



Corresponding author

Correspondence to Thanos D. Halazonetis.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Notes

Supplementary Figures S1-S6 and Supplementary Methods. (PDF 460 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Gorgoulis, V., Vassiliou, LV., Karakaidos, P. et al. Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions. Nature 434, 907–913 (2005).

Download citation

Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing