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Functional Characterization of HC11 Mouse Mammary Epithelial Cells Transduced with Abcg2
Author(s) -
Wu Alex,
Dalvi Pooja,
Earl Brian,
Yang Mingdong,
Harper Patricia,
Ito Shinya
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.lb528
Subject(s) - abcg2 , matrigel , cell culture , transfection , chemistry , microbiology and biotechnology , cancer research , biology , pharmacology , transporter , angiogenesis , biochemistry , atp binding cassette transporter , gene , genetics
The Breast Cancer Resistance Protein (BCRP/ABCG2) is a multidrug efflux transporter that is highly expressed in certain cancer cells and at biological barriers such as the intestine. ABCG2 is also dramatically upregulated in mammary epithelial cells during lactation where it plays a role in the transfer of drugs and toxins into breast milk. Despite the large number of cell lines that overexpress ABCG2, none are suitable models for ABCG2‐mediated drug excretion into breast milk. We report here the creation of a non‐tumorigenic mouse mammary epithelial cell line that expresses high levels of functional ABCG2. Prolactin‐responsive HC11 mouse mammary epithelial cells, which form globular acinar‐like structures with a central lumen (called mammosphere) in Matrigel®, were transduced with a lentiviral vector encoding mouse Abcg2 . One of the 48 clones isolated (designated HC11.Abcg2.C10) showed high levels of ABCG2. HC11.Abcg2.C10 displayed significantly lower intracellular retention of ABCG2 substrates BODIPY‐Prazosin and Hoechst 33342 compared to parent HC11 cells. We then identified an optimal Matrigel‐based culture condition, which supports these cells to form mammosphere structures. Future work will focus on developing methods to measure drug accumulation in the acinar lumen as a model for ABCG2‐mediated drug excretion into breast milk. This work is supported by the Canadian Institutes of Health Research.