Abstract
The sensitivity of human tumor cells to chemotherapeutic agents has recently been correlated to their ability to eliminate drug-induced DNA damage. However, the exact mechanisms by which specific DNA adducts cause cell death are still not fully understood nor are the relative efficacies of distinct pathways within the complex DNA repair network of mammalian cells. To elucidate the link between repair and drug resistance in primary cancer cells we have measured the functional DNA repair capacity of normal and leukemic human lymphocytes ex vivo. Two sensitive assays (the immuno-cytological assay, ICA, detecting specific DNA adducts, and the comet assay, measuring the kinetics of DNA strand break processing) were used to determine the repair capacity at the single cell level.
The individual kinetics for the removal of primary or secondary lesions from nuclear DNA varied significantly (by factors > 10) between cell specimens derived from both healthy individuals and CLL patients. An inverse correlation was observed between the DNA repair capacity of CLL lymphocytes and their in vitro sensitivity to various mono- and bifunctional alkylating agents.
The application of modifiers which block distinct steps of DNA repair pathways provided information about their relative contribution to damage processing in individual cell samples or in cell subpopulations. Furthermore, this strategy revealed a loss of stringent control of specific repair functions in leukemic cells in comparison to their normal counterparts.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Neidle S, Waring M (1993) Molecular aspects of anticancer drug-DNA interactions. In: Neidle S, Fuller W, Cohen JS (eds) Topics in molecular and structural biology, Macmillan, London
Friedberg EC, Walker GC, Siede W (1995) DNA repair and mutagenesis. ASM Press, Washington
Chaney SG, Sancar S (1996) DNA repair: enzymatic mechanisms and relevance to drug response. J. Natl. Cancer Inst 88: 1346–1360
Genetic instability in cancer (1996). Lindahl T (ed.) Cancer surveys, Vol. 28, CSH Laboratory Press, Cold Spring Harbor
Buschfort C, Müller MR, Seeber S, Rajewsky MF, Thomale J. (1997) DNA excision repair profiles of normal and leukemic human lymphocytes: Functional analysis at the single cell level. Cancer Res. 57: 651–658
Thomale J, Seiler F, Müller MR, Seeber S, Rajewsky MF (1994) Repair of 06-alkylguanines in the nuclear DNA of human lymphocytes and leukemic cells: Analysis at the single-cell level. Brit. J. Cancer, 69: 698–705.
Müller MR, Seiler F, Thomale J, Buschfort C, Rajewsky MF, Seeber S (1994) Capacity of individual CLL lymphocytes and leukemic blast cells for repair of O6-ethylguanine in DNA: relation to chemosensitivity in vitro and treatment outcome. Cancer Res., 54: 4524–4531
Müller MR, Buschfort C, Thomale J, Lensing C, Rajewsky MF, Seeber S. DNA repair and cellular resistance to alkylating agents in chronic lymphocytic leukemia. submitted for publication
Thomale J, Engelbergs J, Seiler F, Rajewsky MF (1996) Monoclonal antibody-based quantification and repair analysis of specific alkylation products in DNA. In: G. Pfeifer (Ed.) Thomale J, Engelbergs J, Seiler F, Rajewsky MF, pp 89–103, Plenum Press New York
Thomale J, Hochleitner K, Rajewsky MF (1994) Differential formation and repair of the muta-genic DNA alkylation product O6-ethylguanine in transcribed and non-transcribed genes of the rat. J. Biol. Chem., 269: 1681–2160.
Müller MR, Thomale J, Lensing C, Rajewsky MF, Seeber S (1993) Chemosensitization to alkylating agents by pentoxifylline, O6-benzylguanine and ethacrynic acid in haematological malignancies. Anticancer Res., 13: 2155–2160.
Karran P, Bignami M (1994) DNA damage tolerance, mismatch repair and genomic instability. Bio Essays 16: 833–839
Liu L, Markowitz S, Gerson SL (1996) Mismatch repair mutations override alkyltransferase in conferring resistance to temozolomide but not to BCNU. Cancer Res. 56: 5375–5379
Fink D, Nebel S, Aebi S, Zheng H, Cenni B, Nehmé A, Christen RD, Howell SB (1996) The role of DNA mismatch repair in platinum drug resistance. Cancer Res. 56: 4881–4886
Bentley P, Thomas A, Thomas J (1996) Failure to induce resistance to cytotoxic drugs in normal lymphocytes. Leuk. Res. 20: 645–648
Ben-Yehuda D, Krichevsky S, Caspi O, Rund D, Pollack A, Abeliovich D, Zelig O, Yaholom V, Paltiel O, Or R, Peretz T, Ben-Neriah S, Yehuda O, Rachmilewitz EA (1996) Microsatellite instability and p53 mutations in therapy-related leukemia suggest mutator phenotype. Blood 88: 4298–4303
Tasaka T, Lee S, Spira S, Takeuchi S, Hatta Y, Nagai M, Takahara J, Koeffler HP (1996) Infrequent microsatellite instability during the evolution of the myelodysplastic syndrome to acute myelocytic leukemia. Leukemia Res. 20: 113–117
Gartenhaus R, Johns MM, Wang P, Rai K, Sidransky D (1996) Mutator phenotype in a subset of chronic lymphocytic leukemia. Blood, 87: 38–41
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Springer-Verlag Berlin Heidelberg
About this paper
Cite this paper
Thomale, J., Müller, M.R., Buschfort, C., Seeber, S., Rajewsky, M.F. (1998). Alterations in DNA Repair: Implications for Leukemia Cell Biology. In: Hiddemann, W., et al. Acute Leukemias VII. Haematology and Blood Transfusion / Hämatologie und Bluttransfusion, vol 39. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-71960-8_1
Download citation
DOI: https://doi.org/10.1007/978-3-642-71960-8_1
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-71962-2
Online ISBN: 978-3-642-71960-8
eBook Packages: Springer Book Archive