ABSTRACT Tumorigenesis is a clonal evolution process that is initiated from single cells within otherwise histologically normal tissue1. It is unclear how single, sporadic mutant cells that

have sustained oncogenic alterations evolve within a tightly regulated tissue environment. Here we investigated the effects of inducing oncogene expression in single cells in organotypic

mammary acini as a model to elucidate the processes by which oncogenic alterations initiate clonal progression from organized epithelial environments. Sporadic cells induced to overexpress

oncogenes that specifically perturb cell-cycle checkpoints (for example, E7 from human papilloma virus 16, and cyclin D1), deregulate Myc transcription or activate AKT signalling remained

extracellular-regulated kinase and matrix metalloproteinase activities, and genetic alterations that perturb local cell–matrix adhesion drove cell translocation. We also provide evidence

that luminal cell translocation may drive clonal selection by promoting either the death or the expansion of quiescent oncogene-expressing cells, depending on whether the pre-existing

alterations allow anchorage-independent survival and growth. Our data show that the initial outgrowth of single oncogene-expressing cells from organized epithelial structures is a highly

regulated process, and we propose that a cell translocation mechanism allows sporadic mutant cells to evade suppressive micro-environments and elicits clonal selection for survival and

proliferative expansion outside the native niches of these cells. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution

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references ACKNOWLEDGEMENTS We thank S. Valastyan, T. Muranen and W. Lee for critical reading of the manuscript. We thank the members of the Brugge laboratory for comments and discussion,

the Nikon Imaging Center at Harvard Medical School for providing imaging equipment and software, and the laboratory of G. Danuser for imaging software support. This work was supported by a

grant from the National Cancer Institute (CA080111, to J.S.B.) and an American Cancer Society postdoctoral fellowship (C.T.L.). AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of

Cell Biology, Harvard Medical School, Boston, 02115, Massachusetts, USA Cheuk T. Leung & Joan S. Brugge Authors * Cheuk T. Leung View author publications You can also search for this

author inPubMed Google Scholar * Joan S. Brugge View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS C.T.L. conceived the study, performed the

experiments, analysed the data and drafted the manuscript. J.S.B. supervised the study and edited the manuscript. CORRESPONDING AUTHOR Correspondence to Joan S. Brugge. ETHICS DECLARATIONS

COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION This file contains Supplementary Figures 1-16 with legends,

Legends for Supplementary Movies 1-2 and Supplementary Tables 1-8. (PDF 1848 kb) SUPPLEMENTARY MOVIE 1 This file shows a movie of single ErbB2 or GFP control cells within Day16 MCF10A acini

(see Supplementary Information file for full legend). (MOV 1825 kb) SUPPLEMENTARY MOVIE 2 This file shows a movie of single ErbB2 cell translocation and first division in the lumen (see

Supplementary Information file for full legend). (MOV 1053 kb) POWERPOINT SLIDES POWERPOINT SLIDE FOR FIG. 1 POWERPOINT SLIDE FOR FIG. 2 POWERPOINT SLIDE FOR FIG. 3 POWERPOINT SLIDE FOR FIG.

4 RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Leung, C., Brugge, J. Outgrowth of single oncogene-expressing cells from suppressive epithelial

environments. _Nature_ 482, 410–413 (2012). https://doi.org/10.1038/nature10826 Download citation * Received: 03 May 2011 * Accepted: 03 January 2012 * Published: 08 February 2012 * Issue

Date: 16 February 2012 * DOI: https://doi.org/10.1038/nature10826 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a

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