Rapid SINE-mediated detection of cisplatin

DNA adduct formation in vitro and in vivo in blood

Guichun Wang, Lance M. Hallberg, Ella Englander

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Cisplatin (cis-dichlorodiammine platinum II) is one of the most effective antitumor agents to date. Its usefulness is limited, however, by toxicity to healthy tissues, most notably, its nephrotoxicity. To maximize the chemotherapeutic potential of cisplatin and minimize its adverse effects, it is imperative to monitor formation of cisplatin:DNA adducts throughout treatment. We developed a novel, highly sensitive, SINE (Short Interspersed DNA Element)-mediated, PCR-based assay for detection of cisplatin adducts in vitro and in vivo, in DNA from mouse blood cells. The assay relies on the abundance, dispersion and conservation of SINEs in mammalian genomes. The B1 elements at a copy number of 50,000-80,000 are the most abundant SINEs in the mouse genome. Due to their strong sequence conservation, primers complementary to the B1 consensus sequence anneal to the majority of their targets in the genome and allow simultaneous amplification of long random segments of genomic DNA. Thus, in conjunction with the fact that cisplatin adducts block the progression of thermostable polymerase, B1 element-anchored PCR makes a sensitive tool for assessing the overall integrity of the transcribed regions in the mouse genome. The high sensitivity of the assay allows detection of DNA damage at the low cisplatin dosage of 1-8 mg/kg that is considered as sub-chemotherapeutic in experimental animal models. The sensitivity range therefore, makes this assay suitable for the development of predictive correlation for both, the efficacy of treatment as well as induction of nephrotoxicity. Copyright (C) 1999 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)67-74
Number of pages8
JournalMutation Research - DNA Repair
Volume434
Issue number2
DOIs
StatePublished - Jun 23 1999

Fingerprint

Cisplatin
Blood
Assays
Short Interspersed Nucleotide Elements
Genes
Genome
DNA
Conservation
Polymerase Chain Reaction
Consensus Sequence
Antineoplastic Agents
DNA Damage
Amplification
Toxicity
Blood Cells
Animals
Animal Models
Cells
cisplatin-DNA adduct
In Vitro Techniques

Keywords

  • B1
  • Blood
  • DNA:cisplatin adducts
  • Mouse genome
  • PCR
  • SINEs

ASJC Scopus subject areas

  • Toxicology
  • Genetics
  • Molecular Biology

Cite this

Rapid SINE-mediated detection of cisplatin : DNA adduct formation in vitro and in vivo in blood. / Wang, Guichun; Hallberg, Lance M.; Englander, Ella.

In: Mutation Research - DNA Repair, Vol. 434, No. 2, 23.06.1999, p. 67-74.

Research output: Contribution to journalArticle

@article{00ab87efc6c04689b1445bfacb9f7a92,
title = "Rapid SINE-mediated detection of cisplatin: DNA adduct formation in vitro and in vivo in blood",
abstract = "Cisplatin (cis-dichlorodiammine platinum II) is one of the most effective antitumor agents to date. Its usefulness is limited, however, by toxicity to healthy tissues, most notably, its nephrotoxicity. To maximize the chemotherapeutic potential of cisplatin and minimize its adverse effects, it is imperative to monitor formation of cisplatin:DNA adducts throughout treatment. We developed a novel, highly sensitive, SINE (Short Interspersed DNA Element)-mediated, PCR-based assay for detection of cisplatin adducts in vitro and in vivo, in DNA from mouse blood cells. The assay relies on the abundance, dispersion and conservation of SINEs in mammalian genomes. The B1 elements at a copy number of 50,000-80,000 are the most abundant SINEs in the mouse genome. Due to their strong sequence conservation, primers complementary to the B1 consensus sequence anneal to the majority of their targets in the genome and allow simultaneous amplification of long random segments of genomic DNA. Thus, in conjunction with the fact that cisplatin adducts block the progression of thermostable polymerase, B1 element-anchored PCR makes a sensitive tool for assessing the overall integrity of the transcribed regions in the mouse genome. The high sensitivity of the assay allows detection of DNA damage at the low cisplatin dosage of 1-8 mg/kg that is considered as sub-chemotherapeutic in experimental animal models. The sensitivity range therefore, makes this assay suitable for the development of predictive correlation for both, the efficacy of treatment as well as induction of nephrotoxicity. Copyright (C) 1999 Elsevier Science B.V.",
keywords = "B1, Blood, DNA:cisplatin adducts, Mouse genome, PCR, SINEs",
author = "Guichun Wang and Hallberg, {Lance M.} and Ella Englander",
year = "1999",
month = "6",
day = "23",
doi = "10.1016/S0921-8777(99)00021-X",
language = "English (US)",
volume = "434",
pages = "67--74",
journal = "Mutation Research - DNA Repair",
issn = "0921-8777",
publisher = "Elsevier BV",
number = "2",

}

TY - JOUR

T1 - Rapid SINE-mediated detection of cisplatin

T2 - DNA adduct formation in vitro and in vivo in blood

AU - Wang, Guichun

AU - Hallberg, Lance M.

AU - Englander, Ella

PY - 1999/6/23

Y1 - 1999/6/23

N2 - Cisplatin (cis-dichlorodiammine platinum II) is one of the most effective antitumor agents to date. Its usefulness is limited, however, by toxicity to healthy tissues, most notably, its nephrotoxicity. To maximize the chemotherapeutic potential of cisplatin and minimize its adverse effects, it is imperative to monitor formation of cisplatin:DNA adducts throughout treatment. We developed a novel, highly sensitive, SINE (Short Interspersed DNA Element)-mediated, PCR-based assay for detection of cisplatin adducts in vitro and in vivo, in DNA from mouse blood cells. The assay relies on the abundance, dispersion and conservation of SINEs in mammalian genomes. The B1 elements at a copy number of 50,000-80,000 are the most abundant SINEs in the mouse genome. Due to their strong sequence conservation, primers complementary to the B1 consensus sequence anneal to the majority of their targets in the genome and allow simultaneous amplification of long random segments of genomic DNA. Thus, in conjunction with the fact that cisplatin adducts block the progression of thermostable polymerase, B1 element-anchored PCR makes a sensitive tool for assessing the overall integrity of the transcribed regions in the mouse genome. The high sensitivity of the assay allows detection of DNA damage at the low cisplatin dosage of 1-8 mg/kg that is considered as sub-chemotherapeutic in experimental animal models. The sensitivity range therefore, makes this assay suitable for the development of predictive correlation for both, the efficacy of treatment as well as induction of nephrotoxicity. Copyright (C) 1999 Elsevier Science B.V.

AB - Cisplatin (cis-dichlorodiammine platinum II) is one of the most effective antitumor agents to date. Its usefulness is limited, however, by toxicity to healthy tissues, most notably, its nephrotoxicity. To maximize the chemotherapeutic potential of cisplatin and minimize its adverse effects, it is imperative to monitor formation of cisplatin:DNA adducts throughout treatment. We developed a novel, highly sensitive, SINE (Short Interspersed DNA Element)-mediated, PCR-based assay for detection of cisplatin adducts in vitro and in vivo, in DNA from mouse blood cells. The assay relies on the abundance, dispersion and conservation of SINEs in mammalian genomes. The B1 elements at a copy number of 50,000-80,000 are the most abundant SINEs in the mouse genome. Due to their strong sequence conservation, primers complementary to the B1 consensus sequence anneal to the majority of their targets in the genome and allow simultaneous amplification of long random segments of genomic DNA. Thus, in conjunction with the fact that cisplatin adducts block the progression of thermostable polymerase, B1 element-anchored PCR makes a sensitive tool for assessing the overall integrity of the transcribed regions in the mouse genome. The high sensitivity of the assay allows detection of DNA damage at the low cisplatin dosage of 1-8 mg/kg that is considered as sub-chemotherapeutic in experimental animal models. The sensitivity range therefore, makes this assay suitable for the development of predictive correlation for both, the efficacy of treatment as well as induction of nephrotoxicity. Copyright (C) 1999 Elsevier Science B.V.

KW - B1

KW - Blood

KW - DNA:cisplatin adducts

KW - Mouse genome

KW - PCR

KW - SINEs

UR - http://www.scopus.com/inward/record.url?scp=0033065751&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033065751&partnerID=8YFLogxK

U2 - 10.1016/S0921-8777(99)00021-X

DO - 10.1016/S0921-8777(99)00021-X

M3 - Article

VL - 434

SP - 67

EP - 74

JO - Mutation Research - DNA Repair

JF - Mutation Research - DNA Repair

SN - 0921-8777

IS - 2

ER -