Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancer
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10316/8353 https://doi.org/10.1002/cyto.a.20634 |
Resumo: | 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. © 2008 International Society for Advancement of Cytometry |
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Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancerFlow cytometryBcl-27-AADAnnexin VSCLCViabilityWhen 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. © 2008 International Society for Advancement of Cytometry2008info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/8353http://hdl.handle.net/10316/8353https://doi.org/10.1002/cyto.a.20634engCytometry Part A. 73A:12 (2008) 1165-72Santos, A.Ribeiro, A. B. SarmentoLima, M. C. Pedroso deSimões, S.Moreira, J. N.info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2022-05-25T04:34:03Zoai:estudogeral.uc.pt:10316/8353Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:47:26.966619Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancer |
title |
Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancer |
spellingShingle |
Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancer Santos, A. Flow cytometry Bcl-2 7-AAD Annexin V SCLC Viability |
title_short |
Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancer |
title_full |
Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancer |
title_fullStr |
Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancer |
title_full_unstemmed |
Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancer |
title_sort |
Simultaneous evaluation of viability and Bcl-2 in small-cell lung cancer |
author |
Santos, A. |
author_facet |
Santos, A. Ribeiro, A. B. Sarmento Lima, M. C. Pedroso de Simões, S. Moreira, J. N. |
author_role |
author |
author2 |
Ribeiro, A. B. Sarmento Lima, M. C. Pedroso de Simões, S. Moreira, J. N. |
author2_role |
author author author author |
dc.contributor.author.fl_str_mv |
Santos, A. Ribeiro, A. B. Sarmento Lima, M. C. Pedroso de Simões, S. Moreira, J. N. |
dc.subject.por.fl_str_mv |
Flow cytometry Bcl-2 7-AAD Annexin V SCLC Viability |
topic |
Flow cytometry Bcl-2 7-AAD Annexin V SCLC Viability |
description |
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. © 2008 International Society for Advancement of Cytometry |
publishDate |
2008 |
dc.date.none.fl_str_mv |
2008 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10316/8353 http://hdl.handle.net/10316/8353 https://doi.org/10.1002/cyto.a.20634 |
url |
http://hdl.handle.net/10316/8353 https://doi.org/10.1002/cyto.a.20634 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Cytometry Part A. 73A:12 (2008) 1165-72 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
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1799133752079679488 |