Outgrowth of single oncogene-expressing cells from suppressive epithelial environments

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 quiescent within growth-arrested acini. By contrast, single cells that overexpress ERBB2 initiated a cellular cascade involving cell translocation from the epithelial layer, as well as luminal outgrowth that is characteristic of neoplastic progression in early-stage epithelial tumours. In addition, ERBB2-mediated cell translocation to the lumen was found to depend on 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 options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Single-cell induction of oncogenic alteration in mammary acinar culture.
Figure 2: Mechanisms of cell translocation to the lumen.
Figure 3: Luminal translocation and clonal outgrowth from a suppressive epithelial environment.
Figure 4: Cell translocation elicits clonal selection or outgrowth of quiescent mutant cells.

References

  1. 1

    Nowell, P. C. The clonal evolution of tumor cell populations. Science 194, 23–28 (1976)

    ADS  CAS  Article  Google Scholar 

  2. 2

    Dolberg, D. S. & Bissell, M. J. Inability of Rous sarcoma virus to cause sarcomas in the avian embryo. Nature 309, 552–556 (1984)

    ADS  CAS  Article  Google Scholar 

  3. 3

    Holst, C. R. et al. Methylation of p16INK4a promoters occurs in vivo in histologically normal human mammary epithelia. Cancer Res. 63, 1596–1601 (2003)

    CAS  PubMed  Google Scholar 

  4. 4

    Illmensee, K. & Mintz, B. Totipotency and normal differentiation of single teratocarcinoma cells cloned by injection into blastocysts. Proc. Natl Acad. Sci. USA 73, 549–553 (1976)

    ADS  CAS  Article  Google Scholar 

  5. 5

    Jonason, A. S. et al. Frequent clones of p53-mutated keratinocytes in normal human skin. Proc. Natl Acad. Sci. USA 93, 14025–14029 (1996)

    ADS  CAS  Article  Google Scholar 

  6. 6

    Michaloglou, C. et al. BRAFE600-associated senescence-like cell cycle arrest of human naevi. Nature 436, 720–724 (2005)

    ADS  CAS  Article  Google Scholar 

  7. 7

    Weaver, V. M. et al. Reversion of the malignant phenotype of human breast cells in three-dimensional culture and in vivo by integrin blocking antibodies. J. Cell Biol. 137, 231–245 (1997)

    CAS  Article  Google Scholar 

  8. 8

    Slamon, D. J. et al. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235, 177–182 (1987)

    ADS  CAS  Article  Google Scholar 

  9. 9

    Hebner, C., Weaver, V. M. & Debnath, J. Modeling morphogenesis and oncogenesis in three-dimensional breast epithelial cultures. Annu. Rev. Pathol. 3, 313–339 (2008)

    CAS  Article  Google Scholar 

  10. 10

    Kajita, M. et al. Interaction with surrounding normal epithelial cells influences signalling pathways and behaviour of Src-transformed cells. J. Cell Sci. 123, 171–180 (2010)

    CAS  Article  Google Scholar 

  11. 11

    Hogan, C. et al. Characterization of the interface between normal and transformed epithelial cells. Nature Cell Biol. 11, 460–467 (2009)

    CAS  Article  Google Scholar 

  12. 12

    Pearson, G. W. & Hunter, T. Real-time imaging reveals that noninvasive mammary epithelial acini can contain motile cells. J. Cell Biol. 179, 1555–1567 (2007)

    CAS  Article  Google Scholar 

  13. 13

    Lincoln, D. W., II & Bove, K. The transcription factor Ets-1 in breast cancer. Front. Biosci. 10, 506–511 (2005)

    CAS  Article  Google Scholar 

  14. 14

    Bershadsky, A. D., Balaban, N. Q. & Geiger, B. Adhesion-dependent cell mechanosensitivity. Annu. Rev. Cell Dev. Biol. 19, 677–695 (2003)

    CAS  Article  Google Scholar 

  15. 15

    Gibson, M. C. & Perrimon, N. Extrusion and death of DPP/BMP-compromised epithelial cells in the developing Drosophila wing. Science 307, 1785–1789 (2005)

    ADS  CAS  Article  Google Scholar 

  16. 16

    Li, X., Han, Y. & Xi, R. Polycomb group genes Psc and Su(z)2 restrict follicle stem cell self-renewal and extrusion by controlling canonical and noncanonical Wnt signaling. Genes Dev. 24, 933–946 (2010)

    CAS  Article  Google Scholar 

  17. 17

    Rosenblatt, J., Raff, M. C. & Cramer, L. P. An epithelial cell destined for apoptosis signals its neighbors to extrude it by an actin- and myosin-dependent mechanism. Curr. Biol. 11, 1847–1857 (2001)

    CAS  Article  Google Scholar 

  18. 18

    Davis, M. A., Ireton, R. C. & Reynolds, A. B. A core function for p120-catenin in cadherin turnover. J. Cell Biol. 163, 525–534 (2003)

    CAS  Article  Google Scholar 

  19. 19

    Partanen, J. I., Nieminen, A. I., Makela, T. P. & Klefstrom, J. Suppression of oncogenic properties of c-Myc by LKB1-controlled epithelial organization. Proc. Natl Acad. Sci. USA 104, 14694–14699 (2007)

    ADS  Article  Google Scholar 

  20. 20

    Shen, J. & Dahmann, C. Extrusion of cells with inappropriate Dpp signaling from Drosophila wing disc epithelia. Science 307, 1789–1790 (2005)

    ADS  CAS  Article  Google Scholar 

  21. 21

    Bullen, T. F. et al. Characterization of epithelial cell shedding from human small intestine. Lab. Invest. 86, 1052–1063 (2006)

    CAS  Article  Google Scholar 

  22. 22

    Marshall, T. W., Lloyd, I. E., Delalande, J. M., Nathke, I. & Rosenblatt, J. The tumor suppressor adenomatous polyposis coli controls the direction a cell extrudes from an epithelium. Mol. Biol. Cell 22, 3962–3970 (2011)

    CAS  Article  Google Scholar 

  23. 23

    Brawley, C. & Matunis, E. Regeneration of male germline stem cells by spermatogonial dedifferentiation in vivo. Science 304, 1331–1334 (2004)

    ADS  CAS  Article  Google Scholar 

  24. 24

    Aman, A. & Piotrowski, T. Cell migration during morphogenesis. Dev. Biol. 341, 20–33 (2010)

    CAS  Article  Google Scholar 

  25. 25

    Debnath, J. et al. The role of apoptosis in creating and maintaining luminal space within normal and oncogene-expressing mammary acini. Cell 111, 29–40 (2002)

    CAS  Article  Google Scholar 

  26. 26

    Gunawardane, R. N. et al. Novel role for PDEF in epithelial cell migration and invasion. Cancer Res. 65, 11572–11580 (2005)

    CAS  Article  Google Scholar 

Download 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

Affiliations

Authors

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

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

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Sign up for the Nature Briefing newsletter for a daily update on COVID-19 science.
Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing