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Study of markers of epithelial cells by RT-PCR in real-time detection of circulating tumor cells (CTCs) in mononuclear fraction of peripheral blood in patients with breast cancer during chemotherapy

Grant number: 11/22746-9
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): August 01, 2012
Effective date (End): July 31, 2014
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Fernando Luiz Affonso Fonseca
Grantee:Sarah Rodrigues Marsicano
Home Institution: Faculdade de Medicina do ABC (FMABC). Organização Social de Saúde. Fundação do ABC. Santo André , SP, Brazil


The risk of a subsequent relapse tumor resectable breast cancer - the main factor to indicate the use of adjuvant chemotherapy to a patient is extrapolated from the characteristics of the tumor and not in the detection of residual tumor cells in the patient. This is due to the current absence of complementary methods routinely sensitive enough to detect a very small amount of residual breast tumor (DRM minimal residual disease) in a patient who underwent surgery with curative intent for a primarily unresectable cancer.Evidence in the literature corroborates the detection of minimal residual disease in patients with breast cancer, using various techniques such as immunohistochemistry, immuno-cytochemistry (Schoenfeld et al. 1997, Braun et al., 2000) flow cytometry (FERRERO et al., 1990) and polymerase chain reaction (Manhan et al., 2001, Fonseca et al., 2002). Studies by our group demonstrated that it is possible to monitor patients and detect epithelial antigens (CK 19-9 and Her 2) in patients with breast cancer during chemotherapy using qualitative RT-PCR. These antigens are expressed by epithelial cells from tissues such as breast tumors that are not usually expressed, however, for hematological cells (Manhan et al., 2001, Fonseca et al., 2002).The clinical significance of these findings, however, is currently under intense investigation and then with new tools of molecular biology detection techniques, such as it was suggested the possibility of using quantitative RT-PCR in real time could be improved. In fact, it has been shown that the detection of circulating tumor cells may be useful in screening, prognosis and monitoring of the treatment proposed by the clinic. It is important to note that although quantitative RT-PCR is very sensitive and detects small amounts of tumor cells it is necessary to standardize the markers or antigens to be studied for each type of cancer (melanoma, breast, colon, esophagus, head and neck lung) (XI et al., 2007). These authors compare different markers tumor tissue compared to peripheral blood and found that tumor tissues expressed up to 1000 times more than normal peripheral blood. Besides that, there are reports that you can replace immunohistochemistry by RT-PCR Quantitative real-time verification of gene expression in neoplastic tissues Her 2 breast (Schlemmer et al., 2004).This enables us to study the expression of these markers in peripheral blood mononuclear fraction by RT-PCR Quantitative real-time peripheral blood samples of women with breast cancer. These samples will be obtained during chemotherapy and these findings will also be related with the clinical profile of patients and type of treatment proposed. Thus,we propose a methodology for monitoring the proposed treatment and see how the minimal residual disease in breast cancer behaves characterized by the detection of circulating tumor cells.

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MARSICANO, S. R.; KUNIYOSHI, R. K.; GEHRKE, F. S.; ALVES, B. C. A.; AZZALIS, L. A.; FONSECA, F. L. A. Survinin expression in patients with breast cancer during chemotherapy. TUMOR BIOLOGY, v. 36, n. 5, p. 3441-3445, MAY 2015. Web of Science Citations: 1.

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