Utility of siRNA against Keap1 as a strategy to stimulate a cancer chemopreventive phenotype

Tim W. P. Devling, Christopher D. Lindsay, Lesley I. McLellan, Michael McMahon, John D. Hayes

    Research output: Contribution to journalArticle

    95 Citations (Scopus)

    Abstract

    A duplex 21 nucleotide small interfering RNA (siRNA) against human Keap1 is described that represents a unique class of cancer chemopreventive agent. This siRNA can knockdown Keap1 mRNA and thereby relieve negative regulation of nuclear factor erythroid 2 p45-related factor 2 (Nrf2)-mediated gene expression. The siRNA lowered endogenous Keap1 mRNA to
    Original languageEnglish
    Pages (from-to)7280-7285A
    Number of pages6
    JournalProceedings of the National Academy of Sciences of the United States of America
    Volume102
    Issue number20
    DOIs
    Publication statusPublished - 2005

    Fingerprint

    Small Interfering RNA
    Phenotype
    Neoplasms
    Messenger RNA
    Nucleotides
    Gene Expression

    Keywords

    • Antioxidants
    • Chemoprevention
    • Glutathione
    • Nuclear factor erythroid 2 p45-related factor 2
    • Aldo-keto reductase

    Cite this

    Devling, Tim W. P. ; Lindsay, Christopher D. ; McLellan, Lesley I. ; McMahon, Michael ; Hayes, John D. / Utility of siRNA against Keap1 as a strategy to stimulate a cancer chemopreventive phenotype. In: Proceedings of the National Academy of Sciences of the United States of America. 2005 ; Vol. 102, No. 20. pp. 7280-7285A.
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    abstract = "A duplex 21 nucleotide small interfering RNA (siRNA) against human Keap1 is described that represents a unique class of cancer chemopreventive agent. This siRNA can knockdown Keap1 mRNA and thereby relieve negative regulation of nuclear factor erythroid 2 p45-related factor 2 (Nrf2)-mediated gene expression. The siRNA lowered endogenous Keap1 mRNA to",
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    author = "Devling, {Tim W. P.} and Lindsay, {Christopher D.} and McLellan, {Lesley I.} and Michael McMahon and Hayes, {John D.}",
    note = "dc.publisher: National Academy of Sciences A duplex 21 nucleotide small interfering RNA (siRNA) against human Keap1 is described that represents a unique class of cancer chemopreventive agent. This siRNA can knockdown Keap1 mRNA and thereby relieve negative regulation of nuclear factor erythroid 2 p45-related factor 2 (Nrf2)-mediated gene expression. The siRNA lowered endogenous Keap1 mRNA to <30{\%} of control levels between 24 and 72 h after transfection in human HaCaT keratinocyte cells and was capable of blocking ectopic expression of FLAG-tagged human Keap1 protein but not that of ectopic V5-tagged mouse Keap1 protein. Transfection of human HaCaT cells with Keap1 siRNA markedly enhanced endogenous levels of nuclear factor erythroid 2 p45-related factor 2 (Nrf2) protein and increased transcription of an antioxidant response element-driven reporter gene by 2.3-fold. Furthermore, 48 h after transfection of these cells with Keap1 siRNA, expression of aldo-keto reductase 1C1/2 and the glutamate cysteine ligase catalytic and modifier subunits was elevated between 5- and 14-fold. A modest increase of 3-fold in NAD(P)H:quinone oxidoreductase 1 was also observed. The Keap1 siRNA produced a 1.75-fold increase in intracellular glutathione 48 h after transfection. Thus, antagonism of Keap1 by siRNA can be used to preadapt human cells to oxidative stress without the need to expose them to redox stressors. dc.description.sponsorship: University Research Program of Defense Science and Technology Laboratory Contract CU013-923128 Association of International Cancer Research Grant 02-049 Medical Research Council Cooperative Group Grant G0000281",
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    Utility of siRNA against Keap1 as a strategy to stimulate a cancer chemopreventive phenotype. / Devling, Tim W. P.; Lindsay, Christopher D.; McLellan, Lesley I.; McMahon, Michael; Hayes, John D.

    In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 102, No. 20, 2005, p. 7280-7285A.

    Research output: Contribution to journalArticle

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    T1 - Utility of siRNA against Keap1 as a strategy to stimulate a cancer chemopreventive phenotype

    AU - Devling, Tim W. P.

    AU - Lindsay, Christopher D.

    AU - McLellan, Lesley I.

    AU - McMahon, Michael

    AU - Hayes, John D.

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    KW - Antioxidants

    KW - Chemoprevention

    KW - Glutathione

    KW - Nuclear factor erythroid 2 p45-related factor 2

    KW - Aldo-keto reductase

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    M3 - Article

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    JO - Proceedings of the National Academy of Sciences

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