| High-content screening with siRNA optimizes a cell biological approach to drug discovery: defining the role of P53 activation in the cellular response to anticancer drugs. | |
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MedLine Citation:
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PMID: 15475475 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Deciphering the effects of compounds on molecular events within living cells is becoming an increasingly important component of drug discovery. In a model application of the industrial drug discovery process, the authors profiled a panel of 22 compounds using hierarchical cluster analysis of multiparameter high-content screening measurements from nearly 500,000 cells per microplate. RNAi protein knockdown methodology was used with high-content screening to dissect the effects of 2 anticancer drugs on multiple target activities. Camptothecin activated p53 in A549 lung carcinoma cells pretreated with scrambled siRNA, exhibited concentration-dependent cell cycle blocks, and induced moderate microtubule stabilization. Knockdown of camptothecin-induced p53 protein expression with p53 siRNA inhibited the G1/S blocking activity of the drug and diminished its microtubule-stabilizing activity. Paclitaxel activated p53 protein at low concentrations but exhibited G2/M cell cycle blocking activity at higher concentrations where microtubules were stabilized. In cells treated with p53 siRNA, paclitaxel failed to activate p53 protein, but the knockdown did not have a significant effect on the ability of paclitaxel to stabilize microtubules or induce a G2/M cell cycle block. Thus, this model application of the use of RNAi technology within the context of high-content screening shows the potential to provide massive amounts of combinatorial cell biological information on the temporal and spatial responses that cells mount to treatment by promising therapeutic candidates. |
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Authors:
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Kenneth A Giuliano; Yih-Tai Chen; D Lansing Taylor |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of biomolecular screening Volume: 9 ISSN: 1087-0571 ISO Abbreviation: J Biomol Screen Publication Date: 2004 Oct |
Date Detail:
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Created Date: 2004-10-11 Completed Date: 2005-08-25 Revised Date: 2011-05-23 |
Medline Journal Info:
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Nlm Unique ID: 9612112 Medline TA: J Biomol Screen Country: United States |
Other Details:
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Languages: eng Pagination: 557-68 Citation Subset: IM |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Antineoplastic Agents, Phytogenic
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pharmacology Camptothecin / pharmacology Cell Cycle / drug effects Cluster Analysis DNA / analysis Drug Screening Assays, Antitumor / methods* Fluorescent Antibody Technique Humans Models, Biological Paclitaxel / pharmacology RNA, Small Interfering / metabolism* Tumor Cells, Cultured Tumor Suppressor Protein p53 / drug effects, metabolism* |
| Chemical | |
Reg. No./Substance:
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0/Antineoplastic Agents, Phytogenic; 0/RNA, Small Interfering; 0/Tumor Suppressor Protein p53; 33069-62-4/Paclitaxel; 7689-03-4/Camptothecin; 9007-49-2/DNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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