Quantitation of Lipid Peroxidation Product DNA Adducts in Human Prostate by Tandem Mass Spectrometry: A Method That Mitigates Artifacts

Haoqing Chen, Sesha Krishnamachari, Jingshu Guo, Lihua Yao, Paari Murugan, Christopher J. Weight, Robert J. Turesky

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Reactive oxygen species (ROS) and chronic inflammation contribute to DNA damage of many organs, including the prostate. ROS cause oxidative damage to biomolecules, such as lipids, proteins, and nucleic acids, resulting in the formation of toxic and mutagenic intermediates. Lipid peroxidation (LPO) products covalently adduct to DNA and can lead to mutations. The levels of LPO DNA adducts reported in humans range widely. However, a large proportion of the DNA adducts may be attributed to artifact formation during the steps of isolation and nuclease digestion of DNA. We established a method that mitigates artifacts for most LPO adducts during the processing of DNA. We have applied this methodology to measure LPO DNA adducts in the genome of prostate cancer patients, employing ultrahigh-performance liquid chromatography electrospray ionization ion trap multistage mass spectrometry. Our preliminary data show that DNA adducts of acrolein, 6-hydroxy-1,N2-propano-2′-deoxyguanosine (6-OH-PdG) and 8-hydroxy-1,N2-propano-2′-deoxyguanosine (8-OH-PdG) (4-20 adducts per 107 nucleotides) are more prominent than etheno (ϵ) adducts (<0.5 adducts per 108 nucleotides). This analytical methodology will be used to examine the correlation between oxidative stress, inflammation, and LPO adduct levels in patients with benign prostatic hyperplasia and prostate cancer.

Original languageEnglish (US)
Pages (from-to)1850-1862
Number of pages13
JournalChemical research in toxicology
Volume32
Issue number9
DOIs
StatePublished - Sep 16 2019

Bibliographical note

Funding Information:
This work was supported by R01CA122320 and R01CA220367 (R.J.T.) from the National Cancer Institute and R01ES019564 (R.J.T.) from the National Institute of Environmental Health Sciences. Mass spectrometry was supported by Cancer Center Support Grant CA077598 from the National Cancer Institute, and human biospecimens were supported by the National Center for Advancing Translational Sciences of the National Institutes of Health award number UL1TR000114.

Publisher Copyright:
Copyright © 2019 American Chemical Society.

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