Cisplatin DNA adduct detection and depurination measured by 32P DNA radiolabeling and two-dimensional thin-layer chromatography: A time and concentration study

Nazih Farah, Harley S. Dresner, Kenneth J. Searles, Raz Winiarsky, Mark Moosikasuwan, Antonio Cajigas, Sae Hahm, Jacob J. Steinberg

Research output: Contribution to journalArticle

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Abstract

Platinum-based chemotherapies cause the formation of DNA adducts and have profound effects on DNA. This study measured cis- diamminedichloroplatinum II (cisplatin) DNA adducts by 32P-radiolabeling DNA, enzymatically digesting radiolabeled DNA, separating the formed adducts on two-dimensional thin-layer chromatography, and quantitating the adducts with autoradiography and densitometry. HeLa DNA was incubated with cisplatin at varying concentrations (6.25-325 nM) and times (0 min to 72 hr). Cisplatin rapidly depurinated dGMP and dAMP (90%, 15-min incubation with 325 nM cisplatin). Partial depurination of dGMP (15%) and dAMP (25%) occurred with lower cisplatin concentrations at equal incubation times. A minimum of four new adducts, with relatively rapid migratory patterns, were detected at high cisplatin concentrations with short incubation times. These results indicate that the depurination of DNA correlates with DNA adduct formation and that the quantification of these adducts may be applicable to monitoring tumor and host cell response to cisplatin chemotherapy.

Original languageEnglish (US)
Pages (from-to)314-326
Number of pages13
JournalCancer Investigation
Volume18
Issue number4
StatePublished - 2000

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Thin Layer Chromatography
Cisplatin
DNA
DNA Adducts
Drug Therapy
Densitometry
cisplatin-DNA adduct
Platinum
Autoradiography
Neoplasms

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Cisplatin DNA adduct detection and depurination measured by 32P DNA radiolabeling and two-dimensional thin-layer chromatography : A time and concentration study. / Farah, Nazih; Dresner, Harley S.; Searles, Kenneth J.; Winiarsky, Raz; Moosikasuwan, Mark; Cajigas, Antonio; Hahm, Sae; Steinberg, Jacob J.

In: Cancer Investigation, Vol. 18, No. 4, 2000, p. 314-326.

Research output: Contribution to journalArticle

Farah, N, Dresner, HS, Searles, KJ, Winiarsky, R, Moosikasuwan, M, Cajigas, A, Hahm, S & Steinberg, JJ 2000, 'Cisplatin DNA adduct detection and depurination measured by 32P DNA radiolabeling and two-dimensional thin-layer chromatography: A time and concentration study', Cancer Investigation, vol. 18, no. 4, pp. 314-326.
Farah, Nazih ; Dresner, Harley S. ; Searles, Kenneth J. ; Winiarsky, Raz ; Moosikasuwan, Mark ; Cajigas, Antonio ; Hahm, Sae ; Steinberg, Jacob J. / Cisplatin DNA adduct detection and depurination measured by 32P DNA radiolabeling and two-dimensional thin-layer chromatography : A time and concentration study. In: Cancer Investigation. 2000 ; Vol. 18, No. 4. pp. 314-326.
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