Document Detail


Flow cytometric evaluation of apoptosis and cell viability as a criterion of anti-tumour drug toxicity.
MedLine Citation:
PMID:  12894557     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Determination of the drug concentration required to kill 50% of the tumour cells (EC50) does not take into account the propensity of cells to undergo apoptosis and necrosis. These 2 parameters and the viable cells are here assessed by a flow cytometric (FC) approach using propidium iodide (PI) and FITC-Annexin V staining. MATERIALS AND METHODS: A number of carcinoma cell lines of defined p53 status were exposed to cis-PtII for 24 hours, stained with PI and FITC-Annexin V and analyzed by FC. Unstained viable cells, early apoptotic cells and necrotic cells were scored separately in dual parameter plots of green fluorescence (FITC) against red fluorescence (PI) to generate dose response curves. RESULTS: EC50 values for cell viability were found to be 1-4 times higher than survival data from colony assays resembling data obtained by MTT or Crystal Violet vital dye staining. Percentage apoptosis measured by Annexin V binding was in agreement with microscopic scoring of apoptotic cells after Acridine Orange staining. CONCLUSION: The FC assay described gives a good estimate of cell viability resembling data from vital dye staining assays and provides additional information on apoptosis and necrosis. FC data from Annexin V binding and microscopic scoring after Acridine Orange staining were in excellent agreement.
Authors:
J Michie; J Akudugu; A Binder; C E Van Rensburg; L Böhm
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Anticancer research     Volume:  23     ISSN:  0250-7005     ISO Abbreviation:  Anticancer Res.     Publication Date:    2003 May-Jun
Date Detail:
Created Date:  2003-08-04     Completed Date:  2003-08-28     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8102988     Medline TA:  Anticancer Res     Country:  Greece    
Other Details:
Languages:  eng     Pagination:  2675-9     Citation Subset:  IM    
Affiliation:
Department of Radiation Oncology, Radiobiology Laboratory, Department of Pharmacology, University of Stellenbosch, Faculty of Health, Tygerberg Hospital, P.O. Box 19063, Tygerberg 7505, South Africa.
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MeSH Terms
Descriptor/Qualifier:
Antineoplastic Agents / pharmacology*
Apoptosis / drug effects
Carcinoma, Squamous Cell / drug therapy,  pathology
Cell Survival / drug effects
Drug Screening Assays, Antitumor / methods*
Flow Cytometry / methods*
Humans
Male
Prostatic Neoplasms / drug therapy,  pathology
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Antineoplastic Agents

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


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