Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1995 Apr 25;23(8):1300–1306. doi: 10.1093/nar/23.8.1300

Detection of platinum-DNA adducts by 32P-postlabelling.

F A Blommaert 1, C P Saris 1
PMCID: PMC306853  PMID: 7753620

Abstract

We developed a sensitive 32P-postlabeling method for the detection of bifunctional intrastrand crosslinks d(Pt-GpG) and d(Pt-ApG) in DNA in vitro and in vivo. After enzymatic digestion of DNA the positively charged platinum adducts were purified from unplatinated products, using strong cation exchange chromatography. Subsequently the samples were deplatinated with cyanide, because platinated dinucleotides are very poor substrates for polynucleotide kinase. The excess of cyanide was removed using Sep-pak C18 cartridges, and the resulting dinucleoside monophosphates d(GpG) and d(ApG) were subsequently postlabelled. Analysis of the postlabelling mixture was performed by a combined TLC and HPLC-procedure. Good correlations with existing methods (AAS, immunocytochemistry and ELISA) were found in DNA samples treated in vitro and in vivo with cis- or carboplatin. The detection limit of the assay was 1 adduct/10(7) nucleotides in a 10 micrograms DNA sample.

Full text

PDF
1301

Images in this article

1303Image
on p.1303

Selected References

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

  1. Blommaert F. A., Michael C., Terheggen P. M., Muggia F. M., Kortes V., Schornagel J. H., Hart A. A., den Engelse L. Drug-induced DNA modification in buccal cells of cancer patients receiving carboplatin and cisplatin combination chemotherapy, as determined by an immunocytochemical method: interindividual variation and correlation with disease response. Cancer Res. 1993 Dec 1;53(23):5669–5675. [PubMed] [Google Scholar]
  2. Christian M. C. The current status of new platinum analogs. Semin Oncol. 1992 Dec;19(6):720–733. [PubMed] [Google Scholar]
  3. Eastman A. Activation of programmed cell death by anticancer agents: cisplatin as a model system. Cancer Cells. 1990 Aug-Sep;2(8-9):275–280. [PubMed] [Google Scholar]
  4. Eastman A. Reevaluation of interaction of cis-dichloro(ethylenediamine)platinum(II) with DNA. Biochemistry. 1986 Jul 1;25(13):3912–3915. doi: 10.1021/bi00361a026. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Fichtinger-Schepman A. M., Baan R. A., Luiten-Schuite A., van Dijk M., Lohman P. H. Immunochemical quantitation of adducts induced in DNA by cis-diamminedichloroplatinum (II) and analysis of adduct-related DNA-unwinding. Chem Biol Interact. 1985 Nov;55(3):275–288. doi: 10.1016/s0009-2797(85)80135-6. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Fichtinger-Schepman A. M., van der Veer J. L., den Hartog J. H., Lohman P. H., Reedijk J. Adducts of the antitumor drug cis-diamminedichloroplatinum(II) with DNA: formation, identification, and quantitation. Biochemistry. 1985 Jan 29;24(3):707–713. doi: 10.1021/bi00324a025. [DOI] [PubMed] [Google Scholar]
  9. Försti A., Hemminki K. A 32P-postlabelling assay for DNA adducts induced by cis-diamminedichloroplatinum(II). Cancer Lett. 1994 Aug 15;83(1-2):129–137. doi: 10.1016/0304-3835(94)90309-3. [DOI] [PubMed] [Google Scholar]
  10. Gately D. P., Howell S. B. Cellular accumulation of the anticancer agent cisplatin: a review. Br J Cancer. 1993 Jun;67(6):1171–1176. doi: 10.1038/bjc.1993.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grimaldi K. A., McAdam S. R., Souhami R. L., Hartley J. A. DNA damage by anti-cancer agents resolved at the nucleotide level of a single copy gene: evidence for a novel binding site for cisplatin in cells. Nucleic Acids Res. 1994 Jun 25;22(12):2311–2317. doi: 10.1093/nar/22.12.2311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hemminki K., Peltonen K., Mustonen R. 32P-postlabelling of 7-methyl-dGMP ring-opened 7-methyl-dGMP and platinated dGpdG. Chem Biol Interact. 1990;74(1-2):45–54. doi: 10.1016/0009-2797(90)90057-t. [DOI] [PubMed] [Google Scholar]
  13. Hickman J. A. Apoptosis induced by anticancer drugs. Cancer Metastasis Rev. 1992 Sep;11(2):121–139. doi: 10.1007/BF00048059. [DOI] [PubMed] [Google Scholar]
  14. Jennerwein M. M., Eastman A. A polymerase chain reaction-based method to detect cisplatin adducts in specific genes. Nucleic Acids Res. 1991 Nov 25;19(22):6209–6214. doi: 10.1093/nar/19.22.6209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lemaire M. A., Schwartz A., Rahmouni A. R., Leng M. Interstrand cross-links are preferentially formed at the d(GC) sites in the reaction between cis-diamminedichloroplatinum (II) and DNA. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1982–1985. doi: 10.1073/pnas.88.5.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. May A., Nairn R. S., Okumoto D. S., Wassermann K., Stevnsner T., Jones J. C., Bohr V. A. Repair of individual DNA strands in the hamster dihydrofolate reductase gene after treatment with ultraviolet light, alkylating agents, and cisplatin. J Biol Chem. 1993 Jan 25;268(3):1650–1657. [PubMed] [Google Scholar]
  17. Murray V., Motyka H., England P. R., Wickham G., Lee H. H., Denny W. A., McFadyen W. D. An investigation of the sequence-specific interaction of cis-diamminedichloroplatinum(II) and four analogues, including two acridine-tethered complexes, with DNA inside human cells. Biochemistry. 1992 Dec 1;31(47):11812–11817. doi: 10.1021/bi00162a020. [DOI] [PubMed] [Google Scholar]
  18. Oshita F., Eastman A. Gene-specific damage produced by cisplatin, ormaplatin and UV light in human cells as assayed by the polymerase chain reaction. Oncol Res. 1993;5(3):111–118. [PubMed] [Google Scholar]
  19. 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]
  20. Plooy A. C., van Dijk M., Berends F., Lohman P. H. Formation and repair of DNA interstrand cross-links in relation to cytotoxicity and unscheduled DNA synthesis induced in control and mutant human cells treated with cis-diamminedichloroplatinum(II). Cancer Res. 1985 Sep;45(9):4178–4184. [PubMed] [Google Scholar]
  21. 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]
  22. Poirier M. C., Reed E., Zwelling L. A., Ozols R. F., Litterst C. L., Yuspa S. H. Polyclonal antibodies to quantitate cis-diamminedichloroplatinum(II)--DNA adducts in cancer patients and animal models. Environ Health Perspect. 1985 Oct;62:89–94. doi: 10.1289/ehp.856289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Randerath K., Reddy M. V., Gupta R. C. 32P-labeling test for DNA damage. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6126–6129. doi: 10.1073/pnas.78.10.6126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Reed E., Gupta-Burt S., Litterst C. L., Poirier M. C. Characterization of the DNA damage recognized by an antiserum elicited against cis-diamminedichloroplatinum (II)-modified DNA. Carcinogenesis. 1990 Dec;11(12):2117–2121. doi: 10.1093/carcin/11.12.2117. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Terheggen P. M., Floot B. G., Lempers E. L., van Tellingen O., Begg A. C., den Engelse L. Antibodies against cisplatin-modified DNA and cisplatin-modified (di)nucleotides. Cancer Chemother Pharmacol. 1991;28(3):185–191. doi: 10.1007/BF00685507. [DOI] [PubMed] [Google Scholar]
  27. Terheggen P. M., Floot B. G., Scherer E., Begg A. C., Fichtinger-Schepman A. M., den Engelse L. Immunocytochemical detection of interaction products of cis-diamminedichloroplatinum(II) and cis-diammine(1,1-cyclobutanedicarboxylato)platinum(II) with DNA in rodent tissue sections. Cancer Res. 1987 Dec 15;47(24 Pt 1):6719–6725. [PubMed] [Google Scholar]
  28. Van Benthem J., Wild C. P., Vermeulen E., Den Engelse L., Scherer E. Immunocytochemical localization of DNA adducts induced by a single dose of N-nitroso-N-methylbenzylamine in target and non-target tissues of tumor formation in the rat. Carcinogenesis. 1991 Oct;12(10):1831–1837. doi: 10.1093/carcin/12.10.1831. [DOI] [PubMed] [Google Scholar]
  29. de Graeff A., Slebos R. J., Rodenhuis S. Resistance to cisplatin and analogues: mechanisms and potential clinical implications. Cancer Chemother Pharmacol. 1988;22(4):325–332. doi: 10.1007/BF00254240. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES