Assessment of chemotherapy-induced DNA damage in peripheral blood leukocytes of cancer patients using the alkaline comet assay.

The alkaline comet assay was employed to assess the pre- and post-treatment levels of in vivo DNA damage in peripheral blood leukocytes of cancer patients. During the study all patients were given antineoplastic drugs, mainly as polychemotherapy. To quantify the DNA damage, two different comet parameters were evaluated: the tail length and the tail moment. Our results indicate marked interindividual variations between baseline DNA damage in peripheral blood leukocytes recorded among cancer patients prior to the chemotherapy. After intravenous administration of various antineoplastic drugs, a significantly increased level of DNA damage in all cancer patients compared to their pre-treatment values was recorded The highest level of DNA damage was seen following administration of 5-fluorouracil, adriamycin, and cisplatin (FAP protocol). The results indicate that administration of antineoplastic drugs in standard protocols is accompanied by significant DNA damage in peripheral blood leukocytes. In order to diminish the potential risks of developing second neoplasms, a continuous biomonitoring of cancer patients after the ending of chemotherapy becomes important. Despite their limitations, present results confirm the usefulness of the alkaline comet assay as a sensitive biomarker of exposure that enables rapid and simple detection of primary DNA damage in peripheral blood leukocytes of cancer patients. Together with standard cytogenetic endpoints, the comet assay provides a powerful technique for the routine detection of critical DNA lesions produced after administration of antineoplastic drugs in the clinical settings.

[1]  V. Garaj-Vrhovac,et al.  Application of the alkaline comet assay in human biomonitoring for genotoxicity: a study on Croatian medical personnel handling antineoplastic drugs. , 2001, Mutagenesis.

[2]  O. Merk,et al.  Analysis of chromate-induced DNA-protein crosslinks with the comet assay. , 2000, Mutation research.

[3]  R. Pasquini,et al.  Pesticide-induced primary DNA damage in peripheral blood leukocytes of farm workers evaluated by the computerized 'comet' assay. , 2000 .

[4]  R. Sram,et al.  Molecular epidemiology studies on occupational and environmental exposure to mutagens and carcinogens, 1997-1999. , 2000, Environmental health perspectives.

[5]  W. Vogel,et al.  Induced micronucleus frequencies in peripheral lymphocytes as a screening test for carriers of a BRCA1 mutation in breast cancer families. , 2000, Cancer research.

[6]  V. Garaj-Vrhovac,et al.  Application of cytogenetic endpoints and Comet assay on human lymphocytes treated with vincristine in vitro. , 2000, Neoplasma.

[7]  R. Tice,et al.  Single cell gel/comet assay: Guidelines for in vitro and in vivo genetic toxicology testing , 2000, Environmental and molecular mutagenesis.

[8]  A. Collins,et al.  Lack of adverse effect of smoking habit on DNA strand breakage and base damage, as revealed by the alkaline comet assay. , 1999, Mutation research.

[9]  B. Freidlin,et al.  Secondary leukemia or myelodysplastic syndrome after treatment with epipodophyllotoxins. , 1999, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[10]  P. Olive,et al.  DNA damage and repair in individual cells: applications of the comet assay in radiobiology. , 1999, International journal of radiation biology.

[11]  D. McLaren,et al.  The comet assay in clinical practice. , 1999, Acta oncologica.

[12]  S. Piperakis,et al.  Comet assay for nuclear DNA damage. , 1999, Methods in enzymology.

[13]  G. Melino,et al.  Induction of apoptosis by bleomycin in resting and cycling human lymphocytes. , 1998, Mutagenesis.

[14]  W. Dames,et al.  Human effect monitoring in cases of occupational exposure to antineoplastic drugs: a method comparison. , 1998, Occupational and environmental medicine.

[15]  B. Tunca,et al.  Common fragile site expression and genetic predisposition to breast cancer. , 1998, Teratogenesis, carcinogenesis, and mutagenesis.

[16]  R. Sram,et al.  Single cell gel electrophoresis assay: sensitivity of peripheral white blood cells in human population studies. , 1998, Mutagenesis.

[17]  C. Bokemeyer,et al.  Risk of secondary myeloid leukemia and myelodysplastic syndrome following standard-dose chemotherapy or high-dose chemotherapy with stem cell support in patients with potentially curable malignancies , 1998, Journal of Cancer Research and Clinical Oncology.

[18]  J. Bergh,et al.  Alkaline single-cell gel electrophoresis and human biomonitoring for genotoxicity: a pilot study on breast cancer patients undergoing chemotherapy including cyclophosphamide. , 1997, Mutation research.

[19]  K. Szyfter,et al.  Bleomycin-induced DNA damage and its removal in lymphocytes of breast cancer patients studied by comet assay. , 1997, Mutation research.

[20]  I. Fleming,et al.  Second malignant neoplasms occuring in survivors of osteosarcoma , 1997, Cancer.

[21]  Y. Miyamae,et al.  Detection of DNA lesions induced by chemical mutagens by the single cell gel electrophoresis (Comet) assay. 1. Relationship between the onset of DNA damage and the characteristics of mutagens. , 1997, Mutation research.

[22]  R. Wainstok de Calmanovici,et al.  Evaluation of the Porphyrinogenic Risk of Antineoplastics , 1997, Journal of applied toxicology : JAT.

[23]  G. Duménil,et al.  Chemical degradation of wastes of antineoplastic agents: cyclophosphamide, ifosfamide and melphalan , 1996, International archives of occupational and environmental health.

[24]  M. Dusinska,et al.  Comet assay in human biomonitoring studies: Reliability, validation, and applications , 1997, Environmental and molecular mutagenesis.

[25]  C. Edling,et al.  Alkaline single cell gel electrophoresis of DNA fragments in biomonitoring for genotoxicity: an introductory study on healthy human volunteers , 1997, International archives of occupational and environmental health.

[26]  T. Connor,et al.  Monitoring occupational exposure to cancer chemotherapy drugs. , 1996, American journal of health-system pharmacy : AJHP : official journal of the American Society of Health-System Pharmacists.

[27]  G. Hortobagyi,et al.  Treatment-related leukemia in breast cancer patients treated with fluorouracil-doxorubicin-cyclophosphamide combination adjuvant chemotherapy: the University of Texas M.D. Anderson Cancer Center experience. , 1996, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[28]  P. Dhar,et al.  Significance of lymphocytic sister chromatid exchange frequencies in ovarian cancer patients. , 1996, Cancer genetics and cytogenetics.

[29]  M. Falchi,et al.  Analysis of DNA alkylation damage and repair in mammalian cells by the comet assay. , 1996, Mutagenesis.

[30]  P. Olive,et al.  Sizing highly fragmented DNA in individual apoptotic cells using the comet assay and a DNA crosslinking agent. , 1995, Experimental cell research.

[31]  R. C. Garner,et al.  Cyclophosphamide: review of its mutagenicity for an assessment of potential germ cell risks. , 1995, Mutation research.

[32]  B. Trock.  Application of biological markers in cancer environmental epidemiology. , 1995, Toxicology.

[33]  B. Hellman,et al.  The concepts of tail moment and tail inertia in the single cell gel electrophoresis assay. , 1995, Mutation research.

[34]  P. Olive,et al.  The comet assay: a comprehensive review. , 1995, Mutation research.

[35]  G. Speit,et al.  DNA‐damaging effect of cyclophosphamide on human blood cells in vivo and in vitro studied with the single‐cell gel test (comet assay) , 1995, Environmental and molecular mutagenesis.

[36]  S. Parodi,et al.  Sister-chromatid exchanges, chromosomal aberrations and cytotoxicity produced by topoisomerase II-targeted drugs in sensitive (A2780) and resistant (A2780-DX3) human ovarian cancer cells: correlations with the formation of DNA double-strand breaks. , 1994, Mutation research.

[37]  R. Barale,et al.  Microgel electrophoresis assay (comet test) and SCE analysis in human lymphocytes from 100 normal subjects. , 1994, Mutation research.

[38]  A. Schneider,et al.  Cytogenetic studies in lymphocytes of patients with rectal cancer. , 1993, Environmental health perspectives.

[39]  H. Liber,et al.  Mutagenesis after cancer therapy. , 1993, Environmental health perspectives.

[40]  Saito Isao.  Chemistry of drug-induced DNA lesions , 1993 .

[41]  L. Zwelling.  Consequences of the resistance to drug-induced topoisomerase II-mediated DNA cleavage. , 1993, Toxicology letters.

[42]  T. Skopek,et al.  In vivo mutations in human blood cells: biomarkers for molecular epidemiology. , 1993, Environmental health perspectives.

[43]  E. Reed,et al.  Cisplatin-DNA damage and repair in peripheral blood leukocytes in vivo and in vitro. , 1992, Environmental health perspectives.

[44]  D. Rideout,et al.  Synergism and antagonism in cancer chemotherapy , 1992 .

[45]  R. Tice,et al.  High-dose combination alkylating agents with autologous bone-marrow support in patients with breast cancer: preliminary assessment of DNA damage in individual peripheral blood lymphocytes using the single cell gel electrophoresis assay. , 1992, Mutation research.

[46]  A. Eastman,et al.  The origins of DNA breaks: a consequence of DNA damage, DNA repair, or apoptosis? , 1992, Cancer investigation.

[47]  H. Magdelenat,et al.  Genotoxic effects of radiotherapy and chemotherapy on the circulating lymphocytes of breast cancer patients. II. Alteration of DNA repair and chromosome radiosensitivity. , 1990, Mutation research.

[48]  A. Carrano,et al.  International Commission for Protection Against Environmental Mutagens and Carcinogens. ICPEMC publication no. 14. Considerations for population monitoring using cytogenetic techniques. , 1988, Mutation research.

[49]  R. Tice,et al.  A simple technique for quantitation of low levels of DNA damage in individual cells. , 1988, Experimental cell research.

[50]  N. Day,et al.  Risk of leukaemia in ovarian tumour and breast cancer patients following treatment by cyclophosphamide. , 1987, British Journal of Cancer.

[51]  L. Melton,et al.  Melphalan may be a more potent leukemogen than cyclophosphamide. , 1986, Annals of internal medicine.

[52]  I. Barrai.  Problems in the estimation of mutation rates and in the detection of induced mutations in man. , 1975, Mutation research.