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Breast cancer stem cells are involved in Trastuzumab resistance through the HER2 modulation in 3D culture
Author(s) -
Rodríguez Cristina E.,
Berardi Damian E.,
Abrigo Marianela,
Todaro Laura B.,
Bal de Kier Joffé Elisa D.,
Fiszman Gabriel L.
Publication year - 2018
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.26298
Subject(s) - cancer stem cell , cd44 , stem cell , trastuzumab , cancer research , breast cancer , population , skbr3 , biology , flow cytometry , cancer cell , side population , stem cell marker , cancer , immunology , cell , medicine , microbiology and biotechnology , genetics , human breast , environmental health
Abstract Breast cancer human cells culture as spheroids develop autophagy and apoptosis, which promotes Trastuzumab resistance in HER2 overexpressing cells. Our aim was to study the association of the hostile environment developed in 3D with the breast cancer stem cells population and the HER2 modulation. Human mammary adenocarcinoma cell lines were cultured as spheroids using the hanging drop method. We generated hypoxia conditions by using a hypoxic chamber and CoCl 2 treatment. Breast cancer stem cells were measured with mammosphere assays, the analysis of CD44 + CD24 low population by flow cytometry and the pluripotent gene expression by RT‐qPCR. HER2 expression was evaluated by flow cytometry and Western blot. MTS assays were conducted to study cell viability. Hostil environment developed in spheroids, defined by hypoxia and autophagy, modulated the response to Trastuzumab. In HER2+ cells with acquired resistance, we observed an increase in the breast cancer stem cell population. In BT474 spheroids, Trastuzumab induced the acquisition of resistance, along with an increase in breast cancer stem cells. Also, in 3D culture conditions we determined a modulation in the HER2 expression. Moreover, breast cancer stem cells showed enhanced HER2 expression. Finally, cells without HER2 gene amplification cultured as spheroids were sensitive to Trastuzumab, diminishing HER2 expression and cancer stem cells. Our findings show that 3D architecture is able to modulate breast cancer stem cell population and HER2 distribution, modifying the cell response to Trastuzumab.