Linked Life Data

11040064

Source:http://linkedlifedata.com/resource/pubmed/id/11040064

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2000-10-30
pubmed:abstractText
Many tumors overexpress the NQO1 gene, which encodes DT-diaphorase (NADPH:quinone oxidoreductase; EC 1.6.99.2). This obligate two-electron reductase deactivates toxins and activates bioreductive anticancer drugs. We describe the establishment of an isogenic human tumor cell model for DT-diaphorase expression. An expression vector was used in which the human elongation factor 1alpha promoter produces a bicistronic message containing the genes for human NQO1 and puromycin resistance. This was transfected into the human colon BE tumor line, which has a disabling point mutation in NQO1. Two clones, BE2 and BE5, were selected that were shown by immunoblotting and enzyme activity to stably express high levels of DT-diaphorase. Drug response was determined using 96-h exposures compared with the BE vector control. Functional validation of the isogenic model was provided by the much greater sensitivity of the NQO1-transfected cells to the known DT-diaphorase substrates and bioreductive agents streptonigrin (113- to 132-fold) and indoloquinone EO9 (17- to 25-fold) and the inhibition of this potentiation by the DT-diaphorase inhibitor dicoumarol. A lower degree of potentiation was seen with the clinically used agent mitomycin C (6- to 7-fold) and the EO9 analogs, EO7 and EO2, that are poorer substrates for DT-diaphorase (5- to 8-fold and 2- to 3-fold potentiation, respectively), and there was no potentiation or protection with menadione and tirapazamine. Exposure time-dependent potentiation was seen with the diaziquone analogs methyl-diaziquone and RH1 [2, 5-diaziridinyl-3-(hydroxymethyl)-6-methyl-1,4-benzoquinone], the latter being an agent in preclinical development. In contrast to the in vitro potentiation, there was no difference in the response to mitomycin C when BE2 and BE vector control were treated as tumor xenografts in vivo. This isogenic model should be valuable for mechanistic studies and bioreductive drug development.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0026-895X
pubmed:author
pubmed:issnType
Print
pubmed:volume
58
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1146-55
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed-meshheading:11040064-Animals, pubmed-meshheading:11040064-Antibiotics, Antineoplastic, pubmed-meshheading:11040064-Colonic Neoplasms, pubmed-meshheading:11040064-Disease Models, Animal, pubmed-meshheading:11040064-Gene Expression, pubmed-meshheading:11040064-HT29 Cells, pubmed-meshheading:11040064-Humans, pubmed-meshheading:11040064-Mice, pubmed-meshheading:11040064-Mice, Nude, pubmed-meshheading:11040064-Mitomycin, pubmed-meshheading:11040064-Models, Biological, pubmed-meshheading:11040064-NAD(P)H Dehydrogenase (Quinone), pubmed-meshheading:11040064-Neoplasm Transplantation, pubmed-meshheading:11040064-Reducing Agents, pubmed-meshheading:11040064-Streptonigrin, pubmed-meshheading:11040064-Transplantation, Heterologous, pubmed-meshheading:11040064-Treatment Outcome, pubmed-meshheading:11040064-Tumor Cells, Cultured
pubmed:year
2000
pubmed:articleTitle
Establishment of an isogenic human colon tumor model for NQO1 gene expression: application to investigate the role of DT-diaphorase in bioreductive drug activation in vitro and in vivo.
pubmed:affiliation
CRC Centre for Cancer Therapeutics, The Institute of Cancer Research, Sutton, Surrey, United Kingdom.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't