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Master's Dissertation
DOI
10.11606/D.5.2007.tde-03052007-131443
Document
Author
Full name
Yumi Hasegawa Maekawa
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2007
Supervisor
Committee
Pinotti, Jose Aristodemo (President)
Piáto, José Roberto Morales
Prado, Luiz Carlos Batista do
Title in Portuguese
Detecção e caracterização de células epiteliais no sangue periférico de pacientes com carcinoma localmente avançado de mama
Keywords in Portuguese
Citometria de fluxo
Imunocitoquímica
Neoplasia mamária
Queratina
Separação imunomagnética
Abstract in Portuguese
O projeto tem como objetivo a detecção e caracterização das células epiteliais circulantes de pacientes com carcinoma de mama no estádio III. Analisou-se a presença ou ausência destas células antes da realização de quimioterapia neoadjuvante. Para tanto, utilizou-se o sistema de enriquecimento de amostras de sangue periférico por meio da marcação intracelular imunomagnética direta, empregando-se microbeads conjugados com citoqueratina e posterior separação imunomagnética. A amostra final, com as células alvo detectadas, foi então quantificada por imunocitoquímica ou citometria de fluxo. Resultados: A reação de imunocitoquímica no sangue periférico foi negativa em todos os casos enquanto que na citometria de fluxo foi positiva em 22/23 casos analisados. Conclusão: A detecção e caracterização de células tumorais circulantes em pacientes com câncer de mama podem vir a se constituir em um novo fator prognóstico, porém, são ainda necessárias novas estratégias para melhor detectá-las.
Title in English
Detection and characterization of epithelial cells in the peripheral blood of patients with breast cancer
Keywords in English
Breast neoplasms
Flow cytometry
Immunohistochemistry
Immunomagnetic separation
Keratins
Abstract in English
The aim of this study was to detect and characterize carcinoma cells of patients with breast cancer with clinical stage III analyzing the presence or absence before chemotherapy by enrichment of peripheral blood with super paramagnetic microbeads, using direct Immunomagnetic labeling of intracellular 7/8 cytokeratin. For enrichment, magnetically labeled tumor cells are passed over a high-gradient magnetic positive selection column. Cytospin preparations are made after immunomagnetic enrichment and are examined immunocytochemically (ICC) using an anti-cytokeratin and quantified by flow cytometry (CMF). Results: No cytokeratin positive cells were detected in the 32 peripheral blood of patients of breast carcinoma by ICC and cytokeratin positive cells were detected in the 22 of 23 patients by CMF. Conclusion: These methods of detection and characterization breast carcinoma cells will become useful in the diagnosis and prognosis but is necessary optimization of existing methods.
 
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YUMIMestrado2007.pdf (4.11 Mbytes)
Publishing Date
2007-05-17
 
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