| JC-1, a sensitive probe for a simultaneous detection of P-glycoprotein activity and apoptosis in leukemic cells. | |
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MedLine Citation:
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PMID: 16568474 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: JC-1 probe has been successfully used for the analysis of either apoptosis or P-glycoprotein (P-gp) activity. Therefore, we wanted to see if JC-1 could also simultaneously assess both, P-gp activity and apoptosis, in acute myeloid leukemia (AML) cells. METHODS: P-gp activity was measured using JC-1 and compared to the results of the Rhodamine 123 (Rh 123) assay in P-gp negative and P-gp positive cell lines, and 12 AML samples. For apoptosis, spontaneous apoptosis, as well as, apoptosis induced by Cytosine Arabinosine and Homoharringtonine were analyzed. Both mitochondrial red fluorescence and cytoplasmic green fluorescence of JC-1 with and without a P-gp inhibitor (Cyclosporine A : CsA) were used for the identification of apoptotic cells, and this was compared to Annexin V/PI staining. RESULTS: (1) We found a good correlation between JC-1 and Rh 123 in viable cells. Even in a small population of viable cells, P-gp positive cells emitting low red fluorescence, gained on red fluorescence after P-gp inhibition with CsA permitting an evaluation of P-gp activity. (2) We found a good correlation between the Annexin V/PI staining and JC-1 (P < 0.0001) in the assessment of apoptotic cells. Most importantly, the apoptotic cells could be distinguished by the loss of red fluorescence and the increase of green fluorescence without any change after P-gp inhibition with CsA. CONCLUSIONS: JC-1 can simultaneously evaluate two important parameters involved in drug resistance in AML cells, P-gp activity and apoptosis. |
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Authors:
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Driss Chaoui; Anne-Marie Faussat; Patricia Majdak; Ruoping Tang; Jean-Yves Perrot; Sabine Pasco; Christophe Klein; Jean-Pierre Marie; Ollivier Legrand |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Cytometry. Part B, Clinical cytometry Volume: 70 ISSN: 1552-4949 ISO Abbreviation: Cytometry B Clin Cytom Publication Date: 2006 May |
Date Detail:
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Created Date: 2006-04-25 Completed Date: 2006-07-10 Revised Date: 2009-12-11 |
Medline Journal Info:
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Nlm Unique ID: 101235690 Medline TA: Cytometry B Clin Cytom Country: United States |
Other Details:
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Languages: eng Pagination: 189-96 Citation Subset: IM |
Copyright Information:
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Copyright 2006 International Society for Analytical Cytology. |
Affiliation:
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Université Pierre et Marie Curie-Paris 6, UMR S 736, Les Cordeliers, France. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acute Disease Annexin A5 / chemistry Apoptosis* Benzimidazoles / chemistry* Carbocyanines / chemistry* Cell Line, Tumor Cyclosporine / pharmacology Cytoplasm / chemistry, drug effects Flow Cytometry / methods* Fluorescent Dyes / chemistry Humans Leukemia, Myeloid / metabolism, pathology* Mitochondria / chemistry, drug effects P-Glycoprotein / analysis*, antagonists & inhibitors, chemistry Propidium / chemistry Rhodamine 123 / chemistry Staining and Labeling / methods Tumor Cells, Cultured |
| Chemical | |
Reg. No./Substance:
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0/Annexin A5; 0/Benzimidazoles; 0/Carbocyanines; 0/Fluorescent Dyes; 0/P-Glycoprotein; 21527-78-6/5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine; 36015-30-2/Propidium; 59865-13-3/Cyclosporine; 62669-70-9/Rhodamine 123 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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