| Evaluation of a range of anti-proliferative assays for the preclinical screening of anti-psoriatic drugs: a comparison of colorimetric and fluorimetric assays with the thymidine incorporation assay. | |
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MedLine Citation:
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PMID: 20482335 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Established treatments for psoriasis are generally based on antiproliferative, anti-inflammatory, or differentiation-modifying activity, or a combination of these effects. New agents for the treatment of psoriasis could be identified by high-throughput screening (HTS) of large compound libraries using keratinocyte proliferation models. Although several new proliferation assays have been developed, the radioactive [(3)H]-thymidine incorporation assay is still considered to be the gold standard for the evaluation of keratinocyte proliferation in vitro. In this study, we compare a number of simple, and reliable, colorimetric (MTT, NRU, SRB, and CVS), and fluorimetric (CAM and AB) methods with the [(3)H]-thymidine incorporation assay for the measurement of keratinocyte proliferation in the exponential growth phase in 96-well formats. The concentrations that induced 50% growth inhibition (GI(50)) were determined by each assay for the established antipsoriatics, dithranol, and methotrexate. Strong correlations were observed between the percentage growth inhibitions determined by the radioactive and the colorimetric assays, with no significant differences (P > 0.05) between their GI(50) values. The colorimetric assays are thus suitable alternatives to the radioactive assay for quantifying keratinocyte growth inhibition. We have also validated the use of the HaCaT cell line as a representative of the hyperproliferative psoriatic epidermis, in the preclinical screening of experimental anti-psoriatic agents. |
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Authors:
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Suja Elizabeth George; Rosaleen J Anderson; Anne Cunningham; Michael Donaldson; Paul W Groundwater |
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Publication Detail:
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Type: Comparative Study; Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Assay and drug development technologies Volume: 8 ISSN: 1557-8127 ISO Abbreviation: Assay Drug Dev Technol Publication Date: 2010 Jun |
Date Detail:
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Created Date: 2010-06-28 Completed Date: 2010-10-19 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101151468 Medline TA: Assay Drug Dev Technol Country: United States |
Other Details:
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Languages: eng Pagination: 389-400 Citation Subset: IM |
Affiliation:
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Sunderland Pharmacy School, University of Sunderland, Wharncliffe Street, Sunderland, United Kingdom. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Anthralin
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pharmacology Anti-Infective Agents, Local / pharmacology Caspase 3 / metabolism Caspase 7 / metabolism Cell Line Cell Proliferation / drug effects* Colorimetry Dermatologic Agents / pharmacology Dose-Response Relationship, Drug Drug Evaluation, Preclinical / methods* Fluorescent Dyes Fluorometry Humans Keratinocytes / drug effects Methotrexate / pharmacology Psoriasis / drug therapy* Reproducibility of Results Tetrazolium Salts Thiazoles Thymidine / metabolism* |
| Chemical | |
Reg. No./Substance:
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0/Anti-Infective Agents, Local; 0/Dermatologic Agents; 0/Fluorescent Dyes; 0/Tetrazolium Salts; 0/Thiazoles; 298-93-1/thiazolyl blue; 480-22-8/Anthralin; 50-89-5/Thymidine; 59-05-2/Methotrexate; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspase 7 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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