Document Detail

A high-throughput cell-based assay for inhibitors of ABCG2 activity.
MedLine Citation:
PMID:  16490770     Owner:  NLM     Status:  MEDLINE    
ABCG2 is a member of the adenosine triphosphate (ATP)-binding cassette family of multidrug transporters associated with resistance of tumor cells to many cytotoxic agents. Evaluation of modulators of ABCG2 activity has relied on methods such as drug sensitization, biochemical characterization, and transport studies. To search for novel inhibitors of ABCG2, a fluorescent cell-based assay was developed for application in high-throughput screening. Accumulation of pheophorbide a (PhA), an ABCG2-specific substrate, forms the basis for the assay in NCI-H460/MX20 cells overexpressing wild-type ABCG2. Treatment of these cells with 10 microM fumitremorgin C (FTC), a specific ABCG2 inhibitor, increased cell accumulation of PhA to 5.6 times control (Z' 0.5). Validation included confirmation with known ABCG2 inhibitors: FTC, novobiocin, tariquidar, and quercetin. Verapamil, reported to inhibit P-glycoprotein but not ABCG2, had insignificant activity. Screening of a library of 3523 natural products identified 11 compounds with high activity (> or = 50% of FTC, confirmed by reassay), including 3 flavonoids, members of a family of compounds that include ABCG2 inhibitors. One of the inhibitors detected, eupatin, was moderately potent (IC50 of 2.2 microM) and, like FTC, restored sensitivity of resistant cells to mitoxantrone. Application of this assay to other libraries of synthetic compounds and natural products is expected to identify novel inhibitors of ABCG2 activity.
Curtis J Henrich; Heidi R Bokesch; Michael Dean; Susan E Bates; Robert W Robey; Ekaterina I Goncharova; Jennifer A Wilson; James B McMahon
Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural; Research Support, N.I.H., Intramural     Date:  2006-02-20
Journal Detail:
Title:  Journal of biomolecular screening     Volume:  11     ISSN:  1087-0571     ISO Abbreviation:  J Biomol Screen     Publication Date:  2006 Mar 
Date Detail:
Created Date:  2006-03-30     Completed Date:  2006-07-17     Revised Date:  2011-05-23    
Medline Journal Info:
Nlm Unique ID:  9612112     Medline TA:  J Biomol Screen     Country:  United States    
Other Details:
Languages:  eng     Pagination:  176-83     Citation Subset:  IM    
Basic Research Program, SAIC-Frederick, Inc., NCI Frederick, Frederick, MD 21702, USA.
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MeSH Terms
ATP-Binding Cassette Transporters / analysis,  antagonists & inhibitors*
Cell Line, Tumor
Chlorophyll / analogs & derivatives*,  pharmacology
Dose-Response Relationship, Drug
Drug Resistance, Neoplasm
Drug Synergism
Indenes / pharmacology*
Mitoxantrone / pharmacology*
Neoplasm Proteins / analysis,  antagonists & inhibitors*
Radiation-Sensitizing Agents / pharmacology
Grant Support
Reg. No./Substance:
0/ABCG2 protein, human; 0/ATP-Binding Cassette Transporters; 0/Indenes; 0/Neoplasm Proteins; 0/Radiation-Sensitizing Agents; 12626-18-5/Fumitremorgin A; 1406-65-1/Chlorophyll; 15664-29-6/pheophorbide a; 65271-80-9/Mitoxantrone

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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