Breast cancer tissue slices as a model for evaluation of response to rapamycin

Breast cancer tissue slices as a model for evaluation of response to rapamycin

Author Giorgi Grosso, Stana Helena Google Scholar
Hirata Katayama, Maria Lucia Google Scholar
Roela, Rosimeire Aparecida Google Scholar
Nonogaki, Suely Google Scholar
Soares, Fernando Augusto Google Scholar
Brentani, Helena Google Scholar
Lima, Leandro Google Scholar
Azevedo Koike Folgueira, Maria Aparecida Google Scholar
Logullo Waitzberg, Angela Flavia Autor UNIFESP Google Scholar
Pasini, Faima Solange Google Scholar
Guedes Sampaio Goes, Joao Carlos Google Scholar
Mitzi Brentani, M. Google Scholar
Institution Inst Brasileiro Controle Canc
Universidade de São Paulo (USP)
Adolfo Lutz Inst
Hosp A C Camargo
Hosp AC Camargo Fund Antonio Prudente
Universidade Federal de São Paulo (UNIFESP)
Abstract Rapamycin is a selective inhibitor of the mammalian target of rapamycin (mTOR), a regulator kinase that integrates growth factors signaling via the phosphoinositide-3-kinase pathway and that has emerged as a novel therapeutic modality in breast cancer (BC). We propose a pre-clinical ex-vivo personalized organotypic culture of BC that preserves the microenvironment to evaluate rapamycin-mediated gene expression changes. Freshly excised ductal invasive BC slices, 400 mu m thick (n=30), were cultured in the presence or absence (control) of rapamycin (20 nM) for 24 h. Some slices were formalin-fixed for immunohistochemical determinations and some were processed for microarray analysis. Control slices in culture retained their tissue morphology and tissue viability (detected by BrdU uptake). the percentage of proliferating cells (assessed by Ki67) did not change up to 24 h of treatment. Immunohistochemical evaluation of p-AKT, p-mTOR, p-4EBP1 and p-S6K1 indicated that AKT/mTOR pathway activation was maintained during cultivation. for microarray analysis, slices were divided into two groups, according to the presence/absence of epidermal growth factor receptor-type 2 and analyzed separately. Limited overlap was seen among differentially expressed genes after treatment (P < 0.01) in both groups suggesting different responses to rapamycin between these BC subtypes. Ontology analysis indicated that genes involved in biosynthetic processes were commonly reduced by rapamycin. Our network analysis suggested that concerted expression of these genes might distinguish controls from treated slices. Thus, breast carcinoma slices constitute a suitable physiological tool to evaluate the short-term effects of rapamycin on the gene profile of individual BC samples.
Keywords Breast cancer
Ex-vivo model
AKT/mTOR pathway
Co-expression network
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Grant number FAPESP: 2009/10088-7
Date 2013-06-01
Published in Cell and Tissue Research. New York: Springer, v. 352, n. 3, p. 671-684, 2013.
ISSN 0302-766X (Sherpa/Romeo, impact factor)
Publisher Springer
Extent 671-684
Access rights Closed access
Type Article
Web of Science ID WOS:000319427800020

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