Document Detail


A novel reporter system for molecular imaging and high-throughput screening of anticancer drugs.
MedLine Citation:
PMID:  23881799     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Apoptosis is irreversible programmed cell death, characterized by a cellular cascade activation of caspase 3, which subsequently degrades proteins and other components of cells with a motif sequence. Here we report a novel reporter system to detect apoptosis, growth arrest, and cell death based on controlled and self-amplified protein degradation. The key element of the reporter system is an apoptotic sensor chimerical protein which consists of three components: procaspase 3, ubiquitin (Ub), and a strong consensus sequence of N-degron. Between each of these units is a DEVD (Asp-Glu-Val-Asp) sequence, which acts as the cleavage target of caspase 3. This non-conventional signal loss approach is much more sensitive than other native methods that are based on signal gain. The superior sensitivity is demonstrated by its effective application in 386-well high-throughput screening (HTS) with low drug concentrations and a short incubation time. The HTS selection process using this reporter system is very simple and economic. The simplicity eliminates potential errors introduced by multiple steps; there is no need for any substrate. Furthermore, the cells in the assay need not be disrupted, and the morphology of the cells can provide additional information on mechanisms. After HTS, the intact cells can also be used for other analytic analysis. This system thus has a potentially important role in the discovery and development of new anticancer drugs. It also appears to be very versatile, can be used both in vitro and in vivo with different linked reporter genes, and can be used for a variety of imaging applications.
Authors:
Jingping Xie; Chunxia Wang; John Virostko; H Charles Manning; Wellington Pham; Joshua Bauer; John C Gore
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2013-07-24
Journal Detail:
Title:  Chembiochem : a European journal of chemical biology     Volume:  14     ISSN:  1439-7633     ISO Abbreviation:  Chembiochem     Publication Date:  2013 Aug 
Date Detail:
Created Date:  2013-08-14     Completed Date:  2014-03-03     Revised Date:  2014-08-20    
Medline Journal Info:
Nlm Unique ID:  100937360     Medline TA:  Chembiochem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1494-503     Citation Subset:  IM    
Copyright Information:
Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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MeSH Terms
Descriptor/Qualifier:
Antineoplastic Agents / pharmacology*
Apoptosis
Biological Assay
Cell Line
Cell Shape / drug effects
Cell Survival / drug effects
Drug Discovery*
Drug Screening Assays, Antitumor*
Genes, Reporter / drug effects*
Humans
Microscopy, Confocal
Recombinant Proteins / genetics
Reproducibility of Results
Grant Support
ID/Acronym/Agency:
1P50A128323//PHS HHS; P50 CA095103/CA/NCI NIH HHS; P50 CA128323/CA/NCI NIH HHS; U24 CA126588/CA/NCI NIH HHS; U24A126588//PHS HHS
Chemical
Reg. No./Substance:
0/Antineoplastic Agents; 0/Recombinant Proteins
Comments/Corrections

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


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