Regulation of transgenes in three-dimensional cultures of primary mouse mammary cells demonstrates oncogene dependence and identifies cells that survive deinduction Journal Article
| Authors: | Jechlinger, M.; Podsypanina, K.; Varmus, H. |
| Article Title: | Regulation of transgenes in three-dimensional cultures of primary mouse mammary cells demonstrates oncogene dependence and identifies cells that survive deinduction |
| 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-Kras<sup>G12D</sup>;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 Kras<sup>G12D</sup> are induced, the acini enlarge and form solid, depolarized spheres. Upon deinduction of MYC and Kras<sup>G12D</sup> 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. © 2009 by Cold Spring Harbor Laboratory Press. |
| Keywords: | controlled study; genetics; nonhuman; methodology; mitosis; animal cell; mouse; animal; cytology; metabolism; animals; mice; cell viability; cell survival; mus; cell division; apoptosis; transcription initiation; animal experiment; caspase 3; drug effect; physiology; transgenic mouse; mice, transgenic; stem cell; oncogenes; gene expression regulation; oncogene; antiinfective agent; anti-bacterial agents; tumor recurrence; transgene; cell culture techniques; breast epithelium; stem cells; hoe 33342; oncogene k ras; doxycycline; culture technique; mitochondria; breast carcinogenesis; udder; mammary glands, animal; transcriptional activation; mitochondrion; oncogene myc; inducible transgenes; mammary gland; oncogene dependence; primary cells; residual tumor cells; three-dimensional culture; benzimidazole derivative; casp3 protein, mouse; acinar cell; breast cell; concentration response; metaphase; polarization; repolarization; tissue culture cell; mouse mammary tumor virus; benzimidazoles; transgenes |
| Journal Title: | Genes and Development |
| Volume: | 23 |
| Issue: | 14 |
| ISSN: | 0890-9369 |
| Publisher: | Cold Spring Harbor Laboratory Press |
| Date Published: | 2009-07-15 |
| Start Page: | 1677 |
| End Page: | 1688 |
| Language: | English |
| DOI: | 10.1101/gad.1801809 |
| PUBMED: | 19605689 |
| PROVIDER: | scopus |
| PMCID: | PMC2714708 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 2" - "Export Date: 30 November 2010" - "CODEN: GEDEE" - "Source: Scopus" |
Altmetric
Citation Impact
BMJ Impact Analytics
MSK Authors
-
15Podsypanina -
96Varmus
Related MSK Work