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. 1995 Mar;71(3):512–517. doi: 10.1038/bjc.1995.102

Current sample handling methods for measurement of platinum-DNA adducts in leucocytes in man lead to discrepant results in DNA adduct levels and DNA repair.

J Ma 1, J Verweij 1, A S Planting 1, M de Boer-Dennert 1, H E van Ingen 1, M E van der Burg 1, G Stoter 1, J H Schellens 1
PMCID: PMC2033627  PMID: 7880732

Abstract

DNA adduct levels were measured with atomic spectroscopy in white blood cells (WBCs) from patients with solid tumours who were treated with six weekly courses of cisplatin. In 21 patients (I) the WBCs were collected after thawing frozen whole-blood samples according to a previously described method. In 32 other patients (II) WBCs were collected immediately after blood sample collection. The two methods for WBC collection were also compared in vitro. The maximal DNA adduct levels in vivo after the first course were in I 2.48 +/- 1.14 and in II 1.28 +/- 0.40 pg of platinum per microgram of DNA (P < 0.0001). The DNA 'repair' in the first course (DNA adduct level at the end of the infusion minus the level 15 h post infusion) was in I 40% +/- 29% and in II 18% +/- 29% (P = 0.009). These differences were consistent in all measured courses. In vitro, the DNA adduct levels in the freshly prepared WBCs were significantly lower at 0, 1 and 4, but not 24 h, after start of the incubation with cisplatin than in the WBCs collected after freezing and thawing the blood sample. The same experiment with carboplatin in vitro also resulted in significantly lower adducts in freshly isolated WBCs. The higher DNA adduct levels and DNA 'repair' in I are caused by remaining unbound cisplatin in the sample tubes, which can form DNA adducts ex vivo. The same results in vivo can be anticipated when carboplatin is used.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Fichtinger-Schepman A. M., Baan R. A., Berends F. Influence of the degree of DNA modification on the immunochemical determination of cisplatin-DNA adduct levels. Carcinogenesis. 1989 Dec;10(12):2367–2369. doi: 10.1093/carcin/10.12.2367. [DOI] [PubMed] [Google Scholar]
  2. Fichtinger-Schepman A. M., Vendrik C. P., van Dijk-Knijnenburg W. C., de Jong W. H., van der Minnen A. C., Claessen A. M., van der Velde-Visser S. D., de Groot G., Wubs K. L., Steerenberg P. A. Platinum concentrations and DNA adduct levels in tumors and organs of cisplatin-treated LOU/M rats inoculated with cisplatin-sensitive or -resistant immunoglobulin M immunocytoma. Cancer Res. 1989 Jun 1;49(11):2862–2867. [PubMed] [Google Scholar]
  3. Fichtinger-Schepman A. M., van Oosterom A. T., Lohman P. H., Berends F. cis-Diamminedichloroplatinum(II)-induced DNA adducts in peripheral leukocytes from seven cancer patients: quantitative immunochemical detection of the adduct induction and removal after a single dose of cis-diamminedichloroplatinum(II). Cancer Res. 1987 Jun 1;47(11):3000–3004. [PubMed] [Google Scholar]
  4. Fichtinger-Schepman A. M., van der Velde-Visser S. D., van Dijk-Knijnenburg H. C., van Oosterom A. T., Baan R. A., Berends F. Kinetics of the formation and removal of cisplatin-DNA adducts in blood cells and tumor tissue of cancer patients receiving chemotherapy: comparison with in vitro adduct formation. Cancer Res. 1990 Dec 15;50(24):7887–7894. [PubMed] [Google Scholar]
  5. Loehrer P. J., Einhorn L. H. Drugs five years later. Cisplatin. Ann Intern Med. 1984 May;100(5):704–713. doi: 10.7326/0003-4819-100-5-704. [DOI] [PubMed] [Google Scholar]
  6. Ma J., Verweij J., Kolker H. J., van Ingen H. E., Stoter G., Schellens J. H. Pharmacokinetic-dynamic relationship of cisplatin in vitro: simulation of an i.v. bolus and 3 h and 20 h infusion. Br J Cancer. 1994 May;69(5):858–862. doi: 10.1038/bjc.1994.166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Miller S. A., Dykes D. D., Polesky H. F. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988 Feb 11;16(3):1215–1215. doi: 10.1093/nar/16.3.1215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Motzer R. J., Reed E., Perera F., Tang D., Shamkhani H., Poirier M. C., Tsai W. Y., Parker R. J., Bosl G. J. Platinum-DNA adducts assayed in leukocytes of patients with germ cell tumors measured by atomic absorbance spectrometry and enzyme-linked immunosorbent assay. Cancer. 1994 Jun 1;73(11):2843–2852. doi: 10.1002/1097-0142(19940601)73:11<2843::aid-cncr2820731130>3.0.co;2-d. [DOI] [PubMed] [Google Scholar]
  9. Parker R. J., Gill I., Tarone R., Vionnet J. A., Grunberg S., Muggia F. M., Reed E. Platinum-DNA damage in leukocyte DNA of patients receiving carboplatin and cisplatin chemotherapy, measured by atomic absorption spectrometry. Carcinogenesis. 1991 Jul;12(7):1253–1258. doi: 10.1093/carcin/12.7.1253. [DOI] [PubMed] [Google Scholar]
  10. Poirier M. C., Reed E., Shamkhani H., Tarone R. E., Gupta-Burt S. Platinum drug-DNA interactions in human tissues measured by cisplatin-DNA enzyme-linked immunosorbent assay and atomic absorbance spectroscopy. Environ Health Perspect. 1993 Mar;99:149–154. doi: 10.1289/ehp.9399149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Reed E., Litterst C. L., Thill C. C., Yuspa S. H., Poirier M. C. cis-Diamminedichloroplatinum (II)-DNA adduct formation in renal, gonadal, and tumor tissues of male and female rats. Cancer Res. 1987 Feb 1;47(3):718–722. [PubMed] [Google Scholar]
  12. Reed E., Ostchega Y., Steinberg S. M., Yuspa S. H., Young R. C., Ozols R. F., Poirier M. C. Evaluation of platinum-DNA adduct levels relative to known prognostic variables in a cohort of ovarian cancer patients. Cancer Res. 1990 Apr 15;50(8):2256–2260. [PubMed] [Google Scholar]
  13. Reed E., Ozols R. F., Tarone R., Yuspa S. H., Poirier M. C. Platinum-DNA adducts in leukocyte DNA correlate with disease response in ovarian cancer patients receiving platinum-based chemotherapy. Proc Natl Acad Sci U S A. 1987 Jul;84(14):5024–5028. doi: 10.1073/pnas.84.14.5024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Reed E., Ozols R. F., Tarone R., Yuspa S. H., Poirier M. C. The measurement of cisplatin-DNA adduct levels in testicular cancer patients. Carcinogenesis. 1988 Oct;9(10):1909–1911. doi: 10.1093/carcin/9.10.1909. [DOI] [PubMed] [Google Scholar]
  15. Reed E., Parker R. J., Gill I., Bicher A., Dabholkar M., Vionnet J. A., Bostick-Bruton F., Tarone R., Muggia F. M. Platinum-DNA adduct in leukocyte DNA of a cohort of 49 patients with 24 different types of malignancies. Cancer Res. 1993 Aug 15;53(16):3694–3699. [PubMed] [Google Scholar]
  16. Vermorken J. B., van der Vijgh W. J., Klein I., Gall H. E., Pinedo H. M. Pharmacokinetics of free platinum species following rapid, 3-hr and 24-hr infusions of cis-diamminedichloroplatinum (II) and its therapeutic implications. Eur J Cancer Clin Oncol. 1982 Nov;18(11):1069–1074. doi: 10.1016/0277-5379(82)90085-2. [DOI] [PubMed] [Google Scholar]

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