Document Detail

Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancer.
MedLine Citation:
PMID:  18785266     Owner:  NLM     Status:  MEDLINE    
When designing molecular targeted therapeutic strategies against cancer, it is important to correlate protein expression and cell viability. However, such goal can be difficult if performed in separate assays, especially when only a fraction of cells has been efficiently transfected. Therefore, the aim of the present study was to establish a flow cytometry procedure to assess simultaneously Bcl-2 protein level and viability in small-cell lung cancer (SCLC) cells. Viability assessment was performed by staining cells with Annexin V-fluorescein isothiocyanate (FITC) and 7-aminoactinomycin D (7-AAD). Intracellular detection of Bcl-2 was carried out by immunodetection with monoclonal antibodies. Regarding viability determination, the FSC/7-AAD plot identifies the same percentage of viable cells as the FSC/Annexin V-FITC plot, although with greater sensitivity. The procedures involving cells' fixation with 1% paraformaldehyde and permeabilization with digitonin, required for intracellular Bcl-2 immunostaining did not compromise the association of 7-ADD (nor Annexin V-FITC) previously incubated with SCLC cells. It was therefore possible to simultaneously assess cell viability and Bcl-2 protein in SCLC cells. A simple, sensitive, and versatile procedure was established for the first time for the simultaneous evaluation of cell viability and intracellular detection of Bcl-2 in SCLC.
A Santos; A B Sarmento-Ribeiro; M C Pedroso de Lima; S Simões; J N Moreira
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cytometry. Part A : the journal of the International Society for Analytical Cytology     Volume:  73A     ISSN:  1552-4930     ISO Abbreviation:  Cytometry A     Publication Date:  2008 Dec 
Date Detail:
Created Date:  2008-11-21     Completed Date:  2009-08-11     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101235694     Medline TA:  Cytometry A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1165-72     Citation Subset:  IM    
Faculty of Pharmacy, Laboratory of Pharmaceutical Technology, University of Coimbra, Coimbra, Portugal.
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MeSH Terms
Annexin A5 / pharmacology
Antineoplastic Agents / immunology,  pharmacology
Cell Survival
Enzyme Inhibitors / pharmacology
Etoposide / immunology,  pharmacology
Flow Cytometry / methods*
Lung Neoplasms / metabolism*,  pathology
Piperazines / immunology,  pharmacology
Proto-Oncogene Proteins c-bcl-2 / metabolism*
Pyrimidines / immunology,  pharmacology
Small Cell Lung Carcinoma / metabolism*,  pathology
Reg. No./Substance:
0/Annexin A5; 0/Antineoplastic Agents; 0/Enzyme Inhibitors; 0/Piperazines; 0/Proto-Oncogene Proteins c-bcl-2; 0/Pyrimidines; 152459-95-5/imatinib; 33419-42-0/Etoposide

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