@article{f37a21f68e4d4136930167818b882977,
title = "Utility of siRNA against Keap1 as a strategy to stimulate a cancer chemopreventive phenotype",
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 ",
keywords = "Antioxidants, Chemoprevention, Glutathione, Nuclear factor erythroid 2 p45-related factor 2, Aldo-keto reductase",
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 ",
year = "2005",
doi = "10.1073/pnas.0501475102",
language = "English",
volume = "102",
pages = "7280--7285A",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "20",
}
