Flow cytometry in the study of cell death

Bertho, Álvaro Luiz; Santiago, Marta A; Coutinho, S.

doi:10.1590/s0074-02762000000300020

Cited by 43 publications

(23 citation statements)

References 35 publications

(18 reference statements)

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“…Annexin V/PI staining is proved to be sensitive assay for apoptosis detection. Annexin V binds to externalized phosphatidylserine, marker of early apoptosis.PI flows into cells with broken plasma membrane and binds to DNA and RNA [7][8][9]. Apoptosis (programmed cell death) is a crucial process in every cellular system [8,10].…”

Section: Discussionmentioning

confidence: 99%

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Lymphocytes sensitivity to Fas stimulation in healthy and asthmatic children.

Potapińska

Zawadzka-Krajewska²,

Kotuła³

et al. 2010

Folia Histochem Cytobiol.

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Abstract:The T cell hypothesis of asthma is based on the concept that the disease is driven and maintained by the persistence of a specialized subset of chronically activated T memory cells sensitized against an array of allergenic, occupational or viral antigens. Overreaction of CD4+ T cells in the peripheral blood and airway tissues is an invariant feature of asthma; therefore a potent mechanism for augmenting the number of activated T cells in this disease would be the resistance to the normally programmed pathway for cell death. The aim of the study was to evaluate the presence of apoptotic markers on peripheral blood lymphocytes from healthy and asthmatic children before and after stimulation with antiCD95 antibodies. The blood was collected from 21 children with atopic asthma suffering from allergic rhinitis because of house dust mite and/or grass pollen allergens and 8 healthy children matched for their age and sex. Blood was mixed with purified monoclonal antibody antiCD95 (Beckman Coulter), incubated for 24 hours and than stained with Annexin V andPI (Becton Dickinson). Prepared suspensions were analyzed with Cytomics FC 500 (Beckman Coulter) flow cytometer. Annexin V(+)/PI(-) cells were characterized as early apoptotic, Annexin V(+)/PI(+) as late apoptotic and Annexin V(-)/PI(+) as dead. In unstimulated sample from asthmatic children 21.09±11.20% cells were characterized as Annexin V positive/PI negative. After stimulation with antiCD95 Annexin V positive/PI negative cells constituted 18.72±9.42% of cells, p=0.1. In unstimulated sample from healthy children 11.69±6.70% cells were characterized as Annexin V positive/PI negative. In the sample stimulated with antiCD95 16.54±2.98% of cells were Annexin V positive/PI negative, p=0.02. There were no differences between results of late apoptotic and necrotic lymphocytes from healthy and asthmatic children. Performed research indicates that lymphocytes from asthmatic children are resistant to Fas mediated apoptosis.

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Section: Discussionmentioning

confidence: 99%

“…Binding of Annexin V andPI allows to discriminate early, late apoptotic and dead cells. Test can be performed using flow cytometry [4,9,10].…”

Section: Discussionmentioning

confidence: 99%

Lymphocytes sensitivity to Fas stimulation in healthy and asthmatic children.

Potapińska

Zawadzka-Krajewska²,

Kotuła³

et al. 2010

Folia Histochem Cytobiol.

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“…A Citometria de fluxo é um método qualitativo, quantitativo e exato para análise de apoptose (CHINKWO, 2005;BERTHO et al, 2000), porém o ensaio não distingue a apoptose tardia da necrose, pois o método avalia a apoptose pela translocação da fosfotidilserina para a membrana externa fosfolipídica (GRIFFIN et al, 2007;RÜCKER-MARTIN et al, 1999). Entretanto, a avaliação de apoptose por esse método não permite determinar os diferentes estágios de apoptose, denominados precoce e tardia (PEC et al, 2003;BOERSMA et al, 2005).…”

Section: Discussionunclassified

Sinalização celular para apoptose em linhagem celular de adenocarcinoma (MCF-7) e carcinoma ductal invasivo de mama (ZR 7531) tratados com alcalódes isolados de Pterogyne nitens

Duarte¹

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Sinalização celular para apoptose em linhagem celular de adenocarcinoma (MCF-7) e carcinoma ductal invasivo de mama (ZR 7531) tratadas com alcalóides isolados de Pterogyne nitens ARARAQUARA -SPÀ Camila Araki uma pessoa maravilhosa e competente, que esteve sempre presente e auxiliou-me em todos os momentos.Aos membros da banca examinadora, pela disponibilidade e competência na avaliação deste trabalho.À Regina, pela amizade, carinho e atenção, sempre me apoiando.Às queridas amigas Ana Paula, Débora, Rita Karina, que sempre estiveram ao meu lado em todos os momentos, muito obrigadas pela amizade, carinho, incentivo, apoio e por toda ajuda. À todos do laboratório de Citologia e Biologia Celular, estagiários e pós-graduandos, que de uma forma ou de outra contribuíram para a realização deste trabalho Aline, Angélica, Camila Olivato, Claudete, Daniele, Felipe, Heloísa, Isabel, Jamile, Juliana, Mauro, Maura, Rochane.À Elaine e a Tarsia pela imprescendível colaboração durante a realizção deste trabalho, pelo apoio e estímulo, meus afetivos agradecimentos.Ao Ederlan e Delfina pelo auxílio nas etapas finais deste trabalho.À Maria Isabel pelo apoio técnico e competência com que trabalha pelo nosso laboratório, e principalmente, pela amizade, carinho, preocupação, incentivo e pela certeza de sempre poder contar com sua sinceridade.Ao Max, com quem compartilhei momentos de angústias, por sempre me ajudar a ver o lado positivo de todos acontecimentos e com isso ter proporcionado importantes momentos de descontração.Às meninas da pós-graduação Angela, Cláudia, Laura e Sônia pela dedicação e competência com que trabalham pelo nosso curso, pelo acolhimento que nos proporciona e, principalmente, pela amizade e carinho. mama. Para avaliação da apoptose foi observado um intense efeito concentração-resposta em apoptose tardia/necrose e discreto sinal para apoptose precoce em todas concentrações (p<0,01). No ensaio Hoechst/iodeto, observou diferença significante entre os estágios de apoptose precoce e tardia de ambas linhagens. A pteroginina no período t0 e t24, e pteroginidina no período t0 demonstraram possuir intenso efeito concentração resposta para apoptose tardia e discreto efeito para apoptose precoce em todas as concentrações (p<0,01). Entretanto a linhagem MCF-7 tratada com pteroginidina (t24) demonstrou intenso efeito concentração-resposta para necrose (p<0,01). Atividade de caspase 3/7 foi significativamente maior na concentração de 1,125 mM com pteroginina por t0 (p<0,05). Conclusão: Podemos concluir que PGN e PGD possuem efeito anticarcinogênico contra células de câncer de mama in vitro, induzindo a morte celular programada.Palavras-chave: linhagem cancer de mama, atividade citotoxica, alcalóides, apoptose, necrose, citometria de fluxo. ABSTRACTIntroduction: Breast cancer is a major cause of morbidity and mortality among

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“…This is the reason why AO is commonly used to detect apoptosis [37][38]. It is concluded that the binding targets of AO in intact living cells are the various RNAs and acidic vesicles, most of which are lysosomes.…”

Section: Gastric Cancer Cellsmentioning

confidence: 99%

Intraoperative Photodynamic Surgery (iPDS) with Acridine Orange for Musculoskeletal Sarcomas

Kusuzaki¹,

Matsubara²,

Satonaka³

et al. 2014

Cureus

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We recently established the new limb salvage modality of acridine orange (AO) therapy (AOT), in an attempt to develop minimally invasive limb salvage surgery with minimal damage of normal tissues and a low risk of local recurrence. The treatment modality consists of intraoperative photodynamic surgery (iPDS) and photodynamic therapy (iPDT), followed by postoperative radiodynamic therapy (RDT) using AO for patients with high-grade malignant musculoskeletal sarcomas. Clinical results have shown that the treatment is associated with a low risk of local recurrence, the risk being almost the same as that following conventional wide resection, and yields superior limb function as compared to that obtained after wide resection.In this review, we present the detailed mechanism of selective accumulation of AO in sarcomas, which is related to the acidic environment and lysosomal acidity of the tumor cells induced by cancer-specific glycolysis not involving the define tricarboxylic acid (TCA) cycle (Warburg's effect). We also describe the clinical uses of AOT and the procedure for intraoperative photodynamic surgery (iPDS) using local administration of AO.

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Flow cytometry in the study of cell death

Cited by 43 publications

References 35 publications

Lymphocytes sensitivity to Fas stimulation in healthy and asthmatic children.

Lymphocytes sensitivity to Fas stimulation in healthy and asthmatic children.

Sinalização celular para apoptose em linhagem celular de adenocarcinoma (MCF-7) e carcinoma ductal invasivo de mama (ZR 7531) tratados com alcalódes isolados de Pterogyne nitens

Intraoperative Photodynamic Surgery (iPDS) with Acridine Orange for Musculoskeletal Sarcomas

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