Email updates

Keep up to date with the latest news and content from BMC Proceedings and BioMed Central.

This article is part of the supplement: São Paulo Advanced School of Comparative Oncology: Abstracts

Open Access Poster presentation

Characterization of primary mammary epithelial cells with loss of BRCA1 at a single cell level

Rebecca E Nakles1*, Sarah Millman1, M Carla Cabrera1, Peter Johnson13, Susette C Mueller13, Philipp S Hoppe4, Timm Schroeder4 and Priscilla A Furth1235

Author Affiliations

1 Department of Oncology, Georgetown University, Washington, DC, USA

2 Department of Medicine, Georgetown University, Washington, DC, USA

3 Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA

4 Helmholtz Zentrum München – German Research Center for Environmental Health, Ingolstaedter Landstraße 1, D-85764 Neuherberg, Germany

5 Department of Nanobiomedical Science and WCU Research Center of Nanobiomedical Science, Dankook University, Chungnam 330-714, Korea

For all author emails, please log on.

BMC Proceedings 2013, 7(Suppl 2):P58  doi:10.1186/1753-6561-7-S2-P58


The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/1753-6561/7/S2/P58


Published:4 April 2013

© 2013 Nakles et al; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Background

Loss of BRCA1 has been linked to increased cell proliferation in human mammary epithelial cells. To determine if this phenotype is mirrored in the normal-appearing mammary epithelial cells from mouse models of BRCA1 deficiency, time-lapse imaging was performed on primary mammary epithelial cell (PMEC) cultures. Three distinct genetic models were tested to evaluate the role of p53 haploinsufficiency and p53 haploinsufficiency in the background of ERα over-expression to altered cell proliferation.

Methods

PMEC cultures were generated from 10-12 month old wild-type, Brca1f11/f11/MMTV−Cre, Brca1f11/f11/MMTV−Cre/p53+/- and Brca1f11/f11/MMTV−Cre/p53+/−/Tet-op-ER/MMTV-rtTA mice. Live cell phase contrast time-lapse imaging performed for 5-7 days immediately after plating. Timm’s Tracking Tool software (http://www.helmholtz-muenchen.de/scd/service/scientific-services/software-downloads/index.html webcite) was used to measure individual cell lifetimes.

Results

Mean cell lifetimes in generations 1-4 were significantly shorter in PMEC cultures from Brca1f11/f11/MMTV−Cre and Brca1f11/f11/MMTV−Cre/p53+/− mice as compared to Brca1f11/f11/MMTV−Cre/p53+/−/Tet-op-ER/MMTV-rtTA and wild-type mice. A higher percentage of dividing cells were found in Brca1f11/f11/MMTV−Cre, Brca1f11/f11/MMTV−Cre/p53+/− and Brca1f11/f11/MMTV−Cre/p53+/−/Tet-op-ER/MMTV-rtTA mice as compared to wild-type mice. Brca1f11/f11/MMTV−Cre/p53+/−/Tet-op-ER/MMTV-rtTA mice showed the highest level of colony formation and lowest numbers of apoptotic cells. Brca1f11/f11/MMTV−Cre/p53+/− mice showed the lowest level of colony formation and highest number of apoptotic cells.

Conclusions

Loss of Brca1 by itself was sufficient to decrease cell lifetime however this was modifiable by exposure to ERα overexpression. An inverse relationship between colony formation and numbers of apoptotic cells was found. In summary, genotype-specific differences in primary mammary epithelial cell behavior were revealed by single cell tracking.

Acknowledgements

Supported by NCI, NIH RO1CA112176 (P.A.F.), NCI, NIH. R01CA89041-10S1 (P.A.F.), Deutscher Akademischer Austaush Dienst e.V. A/09/72227 Ref. 316 (R.E.N.), Department of Defense W81XWH-11-1-0074 (R.E.N.), NIH IG20 RR025828-01 (Rodent Barrier Facility Equipment), and NIH NCI 5P30CA051008 (Microscopy and Imaging and Animal Shared Resources).