Regulation of transgenes in three-dimensional cultures of primary mouse mammary cells demonstrates oncogene dependence and identifies cells that survive deinduction
- 15 July 2009
- journal article
- Published by Cold Spring Harbor Laboratory in Genes & Development
- Vol. 23 (14), 1677-1688
- https://doi.org/10.1101/gad.1801809
Abstract
The advent of targeted therapies for cancer has provoked interest in experimental models for the systematic study of oncogene dependence. To that end, we developed a three-dimensional (3D) culture system to analyze the responses of primary mouse mammary epithelial cells to the induction and deinduction of oncogenes. Mammary cells derived from normal virgin mice, or from tritransgenic mice (TetO-MYC;TetO-KrasG12D;MMTV-rtTA) in which MYC and mutant Kras can be regulated by doxycycline, develop from single cells into polarized acini. Lumen formation occurs without apparent apoptosis, and the hollow spheres of cells enlarge by division, with metaphase plates oriented perpendicularly to the apical surface. When MYC and KrasG12D are induced, the acini enlarge and form solid, depolarized spheres. Upon deinduction of MYC and KrasG12D the solid structures regress, leaving a repolarized monolayer of viable cells. These cells display a phenotype consistent with progenitors of mammary epithelium: They exclude Hoechst dye 33342, and reform acini in 3D cultures and repopulate mammary fat pads more efficiently than cells harvested from uninduced acini. Moreover, cells in the surviving spheres retain the ability to respond to reinduction and thus may represent the type of cells that give rise to recurrent tumors.Keywords
This publication has 48 references indexed in Scilit:
- Cdc42 controls spindle orientation to position the apical surface during epithelial morphogenesisThe Journal of cell biology, 2008
- Cell-Polarity Dynamics Controls the Mechanism of Lumen Formation in Epithelial MorphogenesisCurrent Biology, 2008
- Oncogene cooperation in tumor maintenance and tumor recurrence in mouse mammary tumors induced by Myc and mutant KrasProceedings of the National Academy of Sciences of the United States of America, 2008
- Real-time imaging reveals that noninvasive mammary epithelial acini can contain motile cellsThe Journal of cell biology, 2007
- Genetic control of single lumen formation in the zebrafish gutNature, 2007
- Generation of a functional mammary gland from a single stem cellNature, 2006
- Purification and unique properties of mammary epithelial stem cellsNature, 2006
- Oncogenes Come of AgeCold Spring Harbor Symposia on Quantitative Biology, 2005
- MYC inactivation uncovers pluripotent differentiation and tumour dormancy in hepatocellular cancerNature, 2004
- Induction of epithelial tubular morphogenesis in vitro by fibroblast-derived soluble factorsCell, 1991