Advertisement

Epithelial Mesenchymal Transition Traits in Human Breast Cancer Cell Lines Parallel the CD44hi/CD24lo/- Stem Cell Phenotype in Human Breast Cancer

  • Tony Blick
  • Honor Hugo
  • Edwin Widodo
  • Mark Waltham
  • Cletus Pinto
  • Sendurai A. Mani
  • Robert A. Weinberg
  • Richard M. Neve
  • Marc E. Lenburg
  • Erik W. ThompsonEmail author
Article

Abstract

We review here the recently emerging relationship between epithelial-mesenchymal transition (EMT) and breast cancer stem cells (BCSC), and provide analyses of published data on human breast cancer cell lines, supporting their utility as a model for the EMT/BCSC state. Genome-wide transcriptional profiling of these cell lines has confirmed the existence of a subgroup with mesenchymal tendencies and enhanced invasive properties (‘Basal B’/Mesenchymal), distinct from subgroups with either predominantly luminal (‘Luminal’) or mixed basal/luminal (‘Basal A’) features (Neve et al. Cancer Cell, 2006). A literature-derived EMT gene signature has shown specific enrichment within the Basal B subgroup of cell lines, consistent with their over-expression of various EMT transcriptional drivers. Basal B cell lines are found to resemble BCSC, being CD44highCD24low. Moreover, gene products that distinguish Basal B from Basal A and Luminal cell lines (Basal B Discriminators) showed close concordance with those that define BCSC isolated from clinical material, as reported by Shipitsin et al. (Cancer Cell, 2007). CD24 mRNA levels varied across Basal B cell lines, correlating with other Basal B Discriminators. Many gene products correlating with CD24 status in Basal B cell lines were also differentially expressed in isolated BCSC. These findings confirm and extend the importance of the cellular product of the EMT with Basal B cell lines, and illustrate the value of analysing these cell lines for new leads that may improve breast cancer outcomes. Gene products specific to Basal B cell lines may serve as tools for the detection, quantification, and analysis of BCSC/EMT attributes.

Keywords

EMT Basal B Mesenchymal Breast cancer Breast cancer stem cell CD24 

Abbreviations

ALDH1

Aldehyde dehydrogenase 1 family, member A1

BCSC

Breast cancer stem cells

BRCA1

Breast cancer 1, early onset

C/EBP β-2

CCAAT/enhancer binding protein (C/EBP), beta

CDH1

E-cadherin

COX-2

Cyclooxygenase-2

CTC

Circulating tumor cells

DDR1

Discoidin domain receptor tyrosine kinase 1

DTC

Disseminated tumor cells

EGF

Epidermal growth factor

EMT

Epithelial-to-mesenchymal transition

EMT-SIG

EMT-signature

EMP3

Epithelial membrane protein 3

ER

Estrogen receptor

EndMT

Endothelial-to-mesenchymal transition

FAK

Focal adhesion kinase

FOSL

Fos-like antigen

GAS6

Growth arrest-specific 6

HEEBO

Human exonic evidence based oligonucleotide array

HOXB7

Homeobox B7

HMLE

Human mammary epithelial cells

HR

Hazard recurrence

MaSC

Mammary stem cells

MEC

Mammary epithelial cells

MET

Mesenchymal-to-epithelial transition

mRNA

Messenger RNA

NFkB

Nuclear factor kappa B

PGI/AMF

Phosphoglucose isomerise/autocrine motility factor

PROCR

Protein C receptor, endothelial (EPCR)

shRNA

Short hairpin ribonucleic acid

Src

Src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (avian)

TGF-beta

Transforming growth factor-beta

Notes

Acknowledgements

The research effort associated with this article was funded in part by the U.S. Army Medical Research and Materiel Command (BC0213201 and BC084667), the Victorian Breast Cancer Research Consortium, The Cancer Council Victoria (#509295) and the National Breast Cancer Foundation (Australia). TB and EWT were supported in part by the Victorian Breast Cancer Research Consortium. HH is supported by a fellowship from the National Breast Cancer Foundation, Australia. EW is the recipient of an AUS Aid Scholarship. Parts of this work were also supported by the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research (Contract DE-AC03-76SF00098) and the California Breast Cancer Research Program (CBCRP) grant # 7FB-0027. SAM lab is supported by V foundations V Scholar award and M. D. Anderson Research Trust Fellow award. RAW is supported in part by the Breast Cancer Research Foundation. The authors are grateful to Dr. Kornelia Polyak for providing prepublication data from the Shipitsin et al. study (2007) for comparative analysis.

Supplementary material

10911_2010_9175_MOESM1_ESM.xls (20 kb)
Supplementary Table 1 The literature pertaining to EMT in breast cancer was searched and molecules shown empirically to cause EMT or change during EMT were assembled. In some cases, additional family members were included. Gene products known to be differentially expressed across different breast cancer cell lines were not included on the basis of that alone. Although not comprehensive, EMT-SIG is an ad hoc, literature-derived gene list from the breast cancer literature. (XLS 20 kb)

References

  1. 1.
    Australian Institute of Health and Welfare. 2006.Google Scholar
  2. 2.
    Australian Institute of Health and Welfare. 2008.Google Scholar
  3. 3.
    American Cancer Society. 2004.Google Scholar
  4. 4.
    Cristofanilli M. The biological information obtainable from circulating tumor cells. Breast. 2009;18 Suppl 3:S38–40.PubMedGoogle Scholar
  5. 5.
    Harris L, Fritsche H, Mennel R, Norton L, Ravdin P, Taube S, et al. American Society of Clinical Oncology 2007 update of recommendations for the use of tumor markers in breast cancer. J Clin Oncol. 2007;25(33):5287–312.PubMedGoogle Scholar
  6. 6.
    Thiery JP, Acloque H, Huang RY, Nieto MA. Epithelial-mesenchymal transitions in development and disease. Cell. 2009;139(5):871–90.PubMedGoogle Scholar
  7. 7.
    Hay ED. An overview of epithelio-mesenchymal transformation. Acta Anat (Basel). 1995;154(1):8–20.Google Scholar
  8. 8.
    Savagner P, Boyer B, Valles AM, Jouanneau J, Thiery JP. Modulations of the epithelial phenotype during embryogenesis and cancer progression. Cancer Treat Res. 1994;71:229–49.PubMedGoogle Scholar
  9. 9.
    Huber MA, Kraut N, Beug H. Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol. 2005;17(5):548–58.PubMedGoogle Scholar
  10. 10.
    Wynn TA. Cellular and molecular mechanisms of fibrosis. J Pathol. 2008;214(2):199–210.PubMedGoogle Scholar
  11. 11.
    Thiery JP. Epithelial-mesenchymal transitions in development and pathologies. Curr Opin Cell Biol. 2003;15(6):740–6.PubMedGoogle Scholar
  12. 12.
    Polyak K, Weinberg RA. Transitions between epithelial and mesenchymal states: acquisition of malignant and stem cell traits. Nat Rev Cancer. 2009;9(4):265–73.PubMedGoogle Scholar
  13. 13.
    Thompson EW, Newgreen DF, Tarin D. Carcinoma invasion and metastasis: a role for epithelial-mesenchymal transition? Cancer Res. 2005;65(14):5991–5. discussion 5995.PubMedGoogle Scholar
  14. 14.
    Hugo H, Ackland ML, Blick T, Lawrence MG, Clements JA, Williams ED, et al. Epithelial–mesenchymal and mesenchymal–epithelial transitions in carcinoma progression. J Cell Physiol. 2007;213(2):374–83.PubMedGoogle Scholar
  15. 15.
    Chaffer CL, Thompson EW, Williams ED. Mesenchymal to epithelial transition in development and disease. Cells Tissues Organs. 2007;185(1–3):7–19.PubMedGoogle Scholar
  16. 16.
    Tsuji T, Ibaragi S, Hu GF. Epithelial-mesenchymal transition and cell cooperativity in metastasis. Cancer Res. 2009;69(18):7135–9.PubMedGoogle Scholar
  17. 17.
    Tarin D, Thompson EW, Newgreen DF. The fallacy of epithelial mesenchymal transition in neoplasia. Cancer Res. 2005;65(14):5996–6000. discussion 6000–1.PubMedGoogle Scholar
  18. 18.
    Christiansen JJ, Rajasekaran AK. Reassessing epithelial to mesenchymal transition as a prerequisite for carcinoma invasion and metastasis. Cancer Res. 2006;66(17):8319–26.PubMedGoogle Scholar
  19. 19.
    Lee JM, Dedhar S, Kalluri R, Thompson EW. The epithelial-mesenchymal transition: new insights in signaling, development, and disease. J Cell Biol. 2006;172(7):973–81.PubMedGoogle Scholar
  20. 20.
    Frisch SM, Screaton RA. Anoikis mechanisms. Curr Opin Cell Biol. 2001;13(5):555–62.PubMedGoogle Scholar
  21. 21.
    Barrallo-Gimeno A, Nieto MA. The Snail genes as inducers of cell movement and survival: implications in development and cancer. Development. 2005;132(14):3151–61.PubMedGoogle Scholar
  22. 22.
    Przybylo JA, Radisky DC. Matrix metalloproteinase-induced epithelial-mesenchymal transition: tumor progression at Snail’s pace. Int J Biochem Cell Biol. 2007;39(6):1082–8.PubMedGoogle Scholar
  23. 23.
    Thomson S, Buck E, Petti F, Griffin G, Brown E, Ramnarine N, et al. Epithelial to mesenchymal transition is a determinant of sensitivity of non-small-cell lung carcinoma cell lines and xenografts to epidermal growth factor receptor inhibition. Cancer Res. 2005;65(20):9455–62.PubMedGoogle Scholar
  24. 24.
    Hugo, Ackland ML, Lawrence MG, Clements JA, Williams ED, and Thompson EW. Epithelial - Mesenchymal and Mesenchymal - Epithelial Transitions in Carcinoma Progression. J Cell Physiol. 2007;213(2):374–83.Google Scholar
  25. 25.
    Klymkowsky MW, Savagner P. Epithelial-mesenchymal transition: a cancer researcher’s conceptual friend and foe. Am J Pathol. 2009;174(5):1588–93.PubMedGoogle Scholar
  26. 26.
    Tomaskovic-Crook E, Thompson EW, Thiery JP. Epithelial to mesenchymal transition and breast cancer. Breast Cancer Res. 2009;11(6):213.PubMedGoogle Scholar
  27. 27.
    Franci C, Takkunen M, Dave N, Alameda F, Gomez S, Rodriguez R, et al. Expression of Snail protein in tumor-stroma interface. Oncogene. 2006;25(37):5134–44.PubMedGoogle Scholar
  28. 28.
    Blanco MJ, Moreno-Bueno G, Sarrio D, Locascio A, Cano A, Palacios J, et al. Correlation of Snail expression with histological grade and lymph node status in breast carcinomas. Oncogene. 2002;21(20):3241–6.PubMedGoogle Scholar
  29. 29.
    Come C, Magnino F, Bibeau F, De Santa Barbara P, Becker KF, Theillet C, et al. Snail and slug play distinct roles during breast carcinoma progression. Clin Cancer Res. 2006;12(18):5395–402.PubMedGoogle Scholar
  30. 30.
    Elloul S, Elstrand MB, Nesland JM, Trope CG, Kvalheim G, Goldberg I, et al. Snail, Slug, and Smad-interacting protein 1 as novel parameters of disease aggressiveness in metastatic ovarian and breast carcinoma. Cancer. 2005;103(8):1631–43.PubMedGoogle Scholar
  31. 31.
    Martin TA, Goyal A, Watkins G, Jiang WG. Expression of the transcription factors snail, slug, and twist and their clinical significance in human breast cancer. Ann Surg Oncol. 2005;12(6):488–96.PubMedGoogle Scholar
  32. 32.
    Chen J, Imanaka N, Griffin JD. Hypoxia potentiates Notch signaling in breast cancer leading to decreased E-cadherin expression and increased cell migration and invasion. Br J Cancer. 2010;102(2):351–60.PubMedGoogle Scholar
  33. 33.
    Moody SE, Perez D, Pan TC, Sarkisian CJ, Portocarrero CP, Sterner CJ, et al. The transcriptional repressor Snail promotes mammary tumor recurrence. Cancer Cell. 2005;8(3):197–209.PubMedGoogle Scholar
  34. 34.
    Trimboli AJ, Fukino K, de Bruin A, Wei G, Shen L, Tanner SM, et al. Direct evidence for epithelial-mesenchymal transitions in breast cancer. Cancer Res. 2008;68(3):937–45.PubMedGoogle Scholar
  35. 35.
    Xue C, Plieth D, Venkov C, Xu C, Neilson EG. The gatekeeper effect of epithelial-mesenchymal transition regulates the frequency of breast cancer metastasis. Cancer Res. 2003;63(12):3386–94.PubMedGoogle Scholar
  36. 36.
    Damonte P, Gregg JP, Borowsky AD, Keister BA, Cardiff RD. EMT tumorigenesis in the mouse mammary gland. Lab Invest. 2007;87(12):1218–26.PubMedGoogle Scholar
  37. 37.
    Ross DT, Perou CM. A comparison of gene expression signatures from breast tumors and breast tissue derived cell lines. Dis Markers. 2001;17(2):99–109.PubMedGoogle Scholar
  38. 38.
    Neve RM, Chin K, Fridlyand J, Yeh J, Baehner FL, Fevr T, et al. A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer Cell. 2006;10(6):515–27.PubMedGoogle Scholar
  39. 39.
    Charafe-Jauffret E, Ginestier C, Monville F, Finetti P, Adelaide J, Cervera N, et al. Gene expression profiling of breast cell lines identifies potential new basal markers. Oncogene. 2006;25(15):2273–84.PubMedGoogle Scholar
  40. 40.
    Thompson EW, Torri J, Sabol M, Sommers CL, Byers S, Valverius EM, et al. Oncogene-induced basement membrane invasiveness in human mammary epithelial cells. Clin Exp Metastasis. 1994;12(3):181–94.PubMedGoogle Scholar
  41. 41.
    Thompson EW, Paik S, Brunner N, Sommers CL, Zugmaier G, Clarke R, et al. Association of increased basement membrane invasiveness with absence of estrogen receptor and expression of vimentin in human breast cancer cell lines. J Cell Physiol. 1992;150(3):534–44.PubMedGoogle Scholar
  42. 42.
    Sommers CL, Byers SW, Thompson EW, Torri JA, Gelmann EP. Differentiation state and invasiveness of human breast cancer cell lines. Breast Cancer Res Treat. 1994;31(2–3):325–35.PubMedGoogle Scholar
  43. 43.
    Gilles C, Polette M, Piette J, Munaut C, Thompson EW, Birembaut P, et al. High level of MT-MMP expression is associated with invasiveness of cervical cancer cells. Int J Cancer. 1996;65(2):209–13.PubMedGoogle Scholar
  44. 44.
    Blick T, Widodo E, Hugo H, Waltham M, Lenburg ME, Neve RM, et al. Epithelial mesenchymal transition traits in human breast cancer cell lines. Clin Exp Metastasis. 2008;25(6):629–42.PubMedGoogle Scholar
  45. 45.
    Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 2008;133(4):704–15.PubMedGoogle Scholar
  46. 46.
    Dumont N, Wilson MB, Crawford YG, Reynolds PA, Sigaroudinia M, Tlsty TD. Sustained induction of epithelial to mesenchymal transition activates DNA methylation of genes silenced in basal-like breast cancers. Proc Natl Acad Sci U S A. 2008;105(39):14867–72.PubMedGoogle Scholar
  47. 47.
    Whitehead RH, Bertoncello I, Webber LM, Pedersen JS. A new human breast carcinoma cell line (PMC42) with stem cell characteristics. I. Morphologic characterization. J Natl Cancer Inst. 1983;70(4):649–61.PubMedGoogle Scholar
  48. 48.
    Ackland ML, Michalczyk A, Whitehead RH. PMC42, A novel model for the differentiated human breast. Exp Cell Res. 2001;263(1):14–22.PubMedGoogle Scholar
  49. 49.
    Ackland ML, Newgreen D, Price JT, Fridman M, Waltham M, Arvanitis A, et al. Epidermal growth factor stimulates epithelio-mesenchymal transition in the stable human breast carcinoma cell line variant PMC42-LA. Lab Invest. 2003;83(3):435–48.PubMedGoogle Scholar
  50. 50.
    Hugo HJ, Wafai R, Blick T, Thompson EW, Newgreen DF. Staurosporine augments EGF-mediated EMT in PMC42-LA cells through actin depolymerisation, focal contact size reduction and Snail1 induction - a model for cross-modulation. BMC Cancer. 2009;9:235–50.PubMedGoogle Scholar
  51. 51.
    Newgreen DF, Minichiello J. Control of epitheliomesenchymal transformation. I. Events in the onset of neural crest cell migration are separable and inducible by protein kinase inhibitors. Dev Biol. 1995;170(1):91–101.PubMedGoogle Scholar
  52. 52.
    Minichiello J, Ben-Ya’acov A, Hearn CJ, Needham B, Newgreen DF. Induction of epithelio-mesenchymal transformation of quail embryonic neural cells by inhibition of atypical protein kinase-C. Cell Tissue Res. 1999;295(2):195–206.PubMedGoogle Scholar
  53. 53.
    Lebret SC, Newgreen DF, Thompson EW, Ackland ML. Induction of epithelial to mesenchymal transition in PMC42-LA human breast carcinoma cells by carcinoma-associated fibroblast secreted factors. Breast Cancer Res. 2007;9(1):R19.PubMedGoogle Scholar
  54. 54.
    Kao J, Salari K, Bocanegra M, Choi YL, Girard L, Gandhi J, et al. Molecular profiling of breast cancer cell lines defines relevant tumor models and provides a resource for cancer gene discovery. PLoS One. 2009;4(7):e6146.PubMedGoogle Scholar
  55. 55.
    Zajchowski DA, Bartholdi MF, Gong Y, Webster L, Liu HL, Munishkin A, et al. Identification of gene expression profiles that predict the aggressive behavior of breast cancer cells. Cancer Res. 2001;61(13):5168–78.PubMedGoogle Scholar
  56. 56.
    Taylor V, Suter U. Epithelial membrane protein-2 and epithelial membrane protein-3: two novel members of the peripheral myelin protein 22 gene family. Gene. 1996;175(1–2):115–20.PubMedGoogle Scholar
  57. 57.
    Zhou W, Jiang Z, Li X, Xu F, Liu Y, Wen P, et al. EMP3 overexpression in primary breast carcinomas is not associated with epigenetic aberrations. J Korean Med Sci. 2009;24(1):97–103.PubMedGoogle Scholar
  58. 58.
    Evtimova V, Zeillinger R, Weidle UH. Identification of genes associated with the invasive status of human mammary carcinoma cell lines by transcriptional profiling. Tumour Biol. 2003;24(4):189–98.PubMedGoogle Scholar
  59. 59.
    Fumoto S, Tanimoto K, Hiyama E, Noguchi T, Nishiyama M, Hiyama K. EMP3 as a candidate tumor suppressor gene for solid tumors. Expert Opin Ther Targets. 2009;13(7):811–22.PubMedGoogle Scholar
  60. 60.
    Varnum BC, Young C, Elliott G, Garcia A, Bartley TD, Fridell YW, et al. Axl receptor tyrosine kinase stimulated by the vitamin K-dependent protein encoded by growth-arrest-specific gene 6. Nature. 1995;373(6515):623–6.PubMedGoogle Scholar
  61. 61.
    Sharif MN, Sosic D, Rothlin CV, Kelly E, Lemke G, Olson EN, et al. Twist mediates suppression of inflammation by type I IFNs and Axl. J Exp Med. 2006;203(8):1891–901.PubMedGoogle Scholar
  62. 62.
    Hafizi S, Dahlback B. Signalling and functional diversity within the Axl subfamily of receptor tyrosine kinases. Cytokine Growth Factor Rev. 2006;17(4):295–304.PubMedGoogle Scholar
  63. 63.
    Zhang YX, Knyazev PG, Cheburkin YV, Sharma K, Knyazev YP, Orfi L, et al. AXL is a potential target for therapeutic intervention in breast cancer progression. Cancer Res. 2008;68(6):1905–15.PubMedGoogle Scholar
  64. 64.
    Gjerdrum C, Tiron C, Hoiby T, Stefansson I, Haugen H, Sandal T, et al. Axl is an essential epithelial-to-mesenchymal transition-induced regulator of breast cancer metastasis and patient survival. Proc Natl Acad Sci U S A. 2010;107(3):1124–9.PubMedGoogle Scholar
  65. 65.
    McCormack O, Chung WY, Fitzpatrick P, Cooke F, Flynn B, Harrison M, et al. Growth arrest-specific gene 6 expression in human breast cancer. Br J Cancer. 2008;98(6):1141–6.Google Scholar
  66. 66.
    Liu L, Greger J, Shi H, Liu Y, Greshock J, Annan R, et al. Novel mechanism of lapatinib resistance in HER2-positive breast tumor cells: activation of AXL. Cancer Res. 2009;69(17):6871–8.PubMedGoogle Scholar
  67. 67.
    Holland SJ, Pan A, Franci C, Hu Y, Chang B, Li W, et al. R428, a selective small molecule inhibitor of axl kinase, blocks tumor spread and prolongs survival in models of metastatic breast cancer. Cancer Res. 2010;70(4):1544–54.PubMedGoogle Scholar
  68. 68.
    Li Y, Ye X, Tan C, Hongo JA, Zha J, Liu J, et al. Axl as a potential therapeutic target in cancer: role of Axl in tumor growth, metastasis and angiogenesis. Oncogene. 2009;28(39):3442–55.PubMedGoogle Scholar
  69. 69.
    Chiappetta G, Ferraro A, Botti G, Monaco M, Pasquinelli R, Vuttariello E, et al. FRA-1 protein overexpression is a feature of hyperplastic and neoplastic breast disorders. BMC Cancer. 2007;7:17–27.PubMedGoogle Scholar
  70. 70.
    Song Y, Song S, Zhang D, Zhang Y, Chen L, Qian L, et al. An association of a simultaneous nuclear and cytoplasmic localization of Fra-1 with breast malignancy. BMC Cancer. 2006;6:298–304.PubMedGoogle Scholar
  71. 71.
    Luo YP, Zhou H, Krueger J, Kaplan C, Liao D, Markowitz D, et al. The role of proto-oncogene Fra-1 in remodeling the tumor microenvironment in support of breast tumor cell invasion and progression. Oncogene. 2010;29(5):662–73.PubMedGoogle Scholar
  72. 72.
    Chen H, Zhu G, Li Y, Padia RN, Dong Z, Pan ZK, et al. Extracellular signal-regulated kinase signaling pathway regulates breast cancer cell migration by maintaining slug expression. Cancer Res. 2009;69(24):9228–35.PubMedGoogle Scholar
  73. 73.
    Shipitsin M, Campbell LL, Argani P, Weremowicz S, Bloushtain-Qimron N, Yao J, et al. Molecular definition of breast tumor heterogeneity. Cancer Cell. 2007;11(3):259–73.PubMedGoogle Scholar
  74. 74.
    Toole BP. Hyaluronan-CD44 Interactions in Cancer: Paradoxes and Possibilities. Clin Cancer Res. 2009;15(24):7462–8.PubMedGoogle Scholar
  75. 75.
    Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF. Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci U S A. 2003;100(7):3983–8.PubMedGoogle Scholar
  76. 76.
    Lynch MD, Cariati M, Purushotham AD. Breast cancer, stem cells and prospects for therapy. Breast Cancer Res. 2006;8(3):211–21.PubMedGoogle Scholar
  77. 77.
    Lim SC, Oh SH. The role of CD24 in various human epithelial neoplasias. Pathol Res Pract. 2005;201(7):479–86.PubMedGoogle Scholar
  78. 78.
    Visvader JE, Lindeman GJ. Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer. 2008;8(10):755–68.PubMedGoogle Scholar
  79. 79.
    Kristiansen G, Sammar M, Altevogt P. Tumour biological aspects of CD24, a mucin-like adhesion molecule. J Mol Histol. 2004;35(3):255–62.PubMedGoogle Scholar
  80. 80.
    Liu R, Wang X, Chen GY, Dalerba P, Gurney A, Hoey T, et al. The prognostic role of a gene signature from tumorigenic breast-cancer cells. N Engl J Med. 2007;356(3):217–26.PubMedGoogle Scholar
  81. 81.
    Al-Hajj M, Clarke MF. Self-renewal and solid tumor stem cells. Oncogene. 2004;23(43):7274–82.PubMedGoogle Scholar
  82. 82.
    Dalerba P, Cho RW, Clarke MF. Cancer stem cells: models and concepts. Annu Rev Med. 2007;58:267–84.PubMedGoogle Scholar
  83. 83.
    Ginestier C, Hur MH, Charafe-Jauffret E, Monville F, Dutcher J, Brown M, et al. ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell. 2007;1(5):555–67.PubMedGoogle Scholar
  84. 84.
    Creighton CJ, Li X, Landis M, Dixon JM, Neumeister VM, Sjolund A, et al. Residual breast cancers after conventional therapy display mesenchymal as well as tumor-initiating features. Proc Natl Acad Sci U S A. 2009;106(33):13820–5.PubMedGoogle Scholar
  85. 85.
    Stingl J, Eirew P, Ricketson I, Shackleton M, Vaillant F, Choi D, et al. Purification and unique properties of mammary epithelial stem cells. Nature. 2006;439(7079):993–7.PubMedGoogle Scholar
  86. 86.
    Trzpis M, McLaughlin PM, de Leij LM, Harmsen MC. Epithelial cell adhesion molecule: more than a carcinoma marker and adhesion molecule. Am J Pathol. 2007;171(2):386–95.PubMedGoogle Scholar
  87. 87.
    Wright MH, Robles AI, Herschkowitz JI, Hollingshead MG, Anver MR, Perou CM, et al. Molecular analysis reveals heterogeneity of mouse mammary tumors conditionally mutant for Brca1. Mol Cancer. 2008;7:29–40.PubMedGoogle Scholar
  88. 88.
    Shackleton M, Vaillant F, Simpson KJ, Stingl J, Smyth GK, Asselin-Labat ML, et al. Generation of a functional mammary gland from a single stem cell. Nature. 2006;439(7072):84–8.PubMedGoogle Scholar
  89. 89.
    Radisky DC, LaBarge MA. Epithelial-mesenchymal transition and the stem cell phenotype. Cell Stem Cell. 2008;2(6):511–2.PubMedGoogle Scholar
  90. 90.
    Morel AP, Lievre M, Thomas C, Hinkal G, Ansieau S, Puisieux A. Generation of breast cancer stem cells through epithelial-mesenchymal transition. PLoS One. 2008;3(8):e2888.PubMedGoogle Scholar
  91. 91.
    Shimono Y, Zabala M, Cho RW, Lobo N, Dalerba P, Qian D, et al. Downregulation of miRNA-200c links breast cancer stem cells with normal stem cells. Cell. 2009;138(3):592–603.PubMedGoogle Scholar
  92. 92.
    Santisteban M, Reiman JM, Asiedu MK, Behrens MD, Nassar A, Kalli KR, et al. Immune-induced epithelial to mesenchymal transition in vivo generates breast cancer stem cells. Cancer Res. 2009;69(7):2887–95.PubMedGoogle Scholar
  93. 93.
    DiMeo TA, Anderson K, Phadke P, Fan C, Perou CM, Naber S, et al. A novel lung metastasis signature links Wnt signaling with cancer cell self-renewal and epithelial-mesenchymal transition in basal-like breast cancer. Cancer Res. 2009;69(13):5364–73.PubMedGoogle Scholar
  94. 94.
    Lim E, Vaillant F, Wu D, Forrest NC, Pal B, Hart AH, et al. Aberrant luminal progenitors as the candidate target population for basal tumor development in BRCA1 mutation carriers. Nat Med. 2009;15(8):907–13.PubMedGoogle Scholar
  95. 95.
    Sarrio D, Rodriguez-Pinilla SM, Hardisson D, Cano A, Moreno-Bueno G, Palacios J. Epithelial-mesenchymal transition in breast cancer relates to the basal-like phenotype. Cancer Res. 2008;68(4):989–97.PubMedGoogle Scholar
  96. 96.
    Logullo AF, Nonogaki S, Pasini FS, Osorio CA, Soares FA, Brentani MM. Concomitant expression of epithelial-mesenchymal transition biomarkers in breast ductal carcinoma: association with progression. Oncol Rep. 2010;23(2):313–20.PubMedGoogle Scholar
  97. 97.
    Herschkowitz JI, Simin K, Weigman VJ, Mikaelian I, Usary J, Hu Z, et al. Identification of conserved gene expression features between murine mammary carcinoma models and human breast tumors. Genome Biol. 2007;8(5):R76.PubMedGoogle Scholar
  98. 98.
    Hennessy BT, Gonzalez-Angulo AM, Stemke-Hale K, Gilcrease MZ, Krishnamurthy S, Lee JS, et al. Characterization of a naturally occurring breast cancer subset enriched in epithelial-to-mesenchymal transition and stem cell characteristics. Cancer Res. 2009;69(10):4116–24.PubMedGoogle Scholar
  99. 99.
    Lien HC, Hsiao YH, Lin YS, Yao YT, Juan HF, Kuo WH, et al. Molecular signatures of metaplastic carcinoma of the breast by large-scale transcriptional profiling: identification of genes potentially related to epithelial-mesenchymal transition. Oncogene. 2007;26(57):7859–71.PubMedGoogle Scholar
  100. 100.
    Mimeault M, Batra SK. Functions of tumorigenic and migrating cancer progenitor cells in cancer progression and metastasis and their therapeutic implications. Cancer Metastasis Rev. 2007;26(1):203–14.PubMedGoogle Scholar
  101. 101.
    Mimeault M, Batra SK. Interplay of distinct growth factors during epithelial mesenchymal transition of cancer progenitor cells and molecular targeting as novel cancer therapies. Ann Oncol. 2007;18(10):1605–19.PubMedGoogle Scholar
  102. 102.
    Brabletz T, Jung A, Spaderna S, Hlubek F, Kirchner T. Opinion: migrating cancer stem cells - an integrated concept of malignant tumour progression. Nat Rev Cancer. 2005;5(9):744–9.PubMedGoogle Scholar
  103. 103.
    Brabletz T, Jung A, Hermann K, Gunther K, Hohenberger W, Kirchner T. Nuclear overexpression of the oncoprotein beta-catenin in colorectal cancer is localized predominantly at the invasion front. Pathol Res Pract. 1998;194(10):701–4.PubMedGoogle Scholar
  104. 104.
    Kokkinos MI, Wafai R, Wong MK, Newgreen DF, Thompson EW, Waltham M. Vimentin and epithelial-mesenchymal transition in human breast cancer-observations in vitro and in vivo. Cells Tissues Organs. 2007;185:191–203.Google Scholar
  105. 105.
    Korsching E, Packeisen J, Liedtke C, Hungermann D, Wulfing P, van Diest PJ, et al. The origin of vimentin expression in invasive breast cancer: epithelial-mesenchymal transition, myoepithelial histogenesis or histogenesis from progenitor cells with bilinear differentiation potential? J Pathol. 2005;206(4):451–7.PubMedGoogle Scholar
  106. 106.
    Li X, Lewis MT, Huang J, Gutierrez C, Osborne CK, Wu MF, et al. Intrinsic resistance of tumorigenic breast cancer cells to chemotherapy. J Natl Cancer Inst. 2008;100(9):672–9.PubMedGoogle Scholar
  107. 107.
    Becker S, Becker-Pergola G, Wallwiener D, Solomayer EF, Fehm T. Detection of cytokeratin-positive cells in the bone marrow of breast cancer patients undergoing adjuvant therapy. Breast Cancer Res Treat. 2006;97(1):91–6.PubMedGoogle Scholar
  108. 108.
    Becker S, Solomayer E, Becker-Pergola G, Wallwiener D, Fehm T. Primary systemic therapy does not eradicate disseminated tumor cells in breast cancer patients. Breast Cancer Res Treat. 2007;106(2):239–43.PubMedGoogle Scholar
  109. 109.
    Braun S, Pantel K, Muller P, Janni W, Hepp F, Kentenich CR, et al. Cytokeratin-positive cells in the bone marrow and survival of patients with stage I, II, or III breast cancer. N Engl J Med. 2000;342(8):525–33.PubMedGoogle Scholar
  110. 110.
    Cristofanilli M, Budd GT, Ellis MJ, Stopeck A, Matera J, Miller MC, et al. Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med. 2004;351(8):781–91.PubMedGoogle Scholar
  111. 111.
    Braun S, Pantel K. Diagnosis and clinical significance of disseminated tumor cells in bone marrow. Dtsch Med Wochenschr. 2000;125(41):1237–9.PubMedGoogle Scholar
  112. 112.
    Hayes DF, Cristofanilli M, Budd GT, Ellis MJ, Stopeck A, Miller MC, et al. Circulating tumor cells at each follow-up time point during therapy of metastatic breast cancer patients predict progression-free and overall survival. Clin Cancer Res. 2006;12(14 Pt 1):4218–24.PubMedGoogle Scholar
  113. 113.
    Wimberger P, Heubner M, Otterbach F, Fehm T, Kimmig R, Kasimir-Bauer S. Influence of platinum-based chemotherapy on disseminated tumor cells in blood and bone marrow of patients with ovarian cancer. Gynecol Oncol. 2007;107(2):331–8.PubMedGoogle Scholar
  114. 114.
    Balic M, Lin H, Young L, Hawes D, Giuliano A, McNamara G, et al. Most early disseminated cancer cells detected in bone marrow of breast cancer patients have a putative breast cancer stem cell phenotype. Clin Cancer Res. 2006;12(19):5615–21.PubMedGoogle Scholar
  115. 115.
    Watson MA, Ylagan LR, Trinkaus KM, Gillanders WE, Naughton MJ, Weilbaecher KN, Fleming TP, Aft RL. Isolation and molecular profiling of bone marrow micrometastases identifies TWIST1 as a marker of early tumor relapse in breast cancer patients. Clin Cancer Res. 2007;13:5001–9.Google Scholar
  116. 116.
    Pantel K, Schlimok G, Angstwurm M, Weckermann D, Schmaus W, Gath H, et al. Methodological analysis of immunocytochemical screening for disseminated epithelial tumor cells in bone marrow. J Hematother. 1994;3(3):165–73.PubMedGoogle Scholar
  117. 117.
    Aktas B, Tewes M, Fehm T, Hauch S, Kimmig R, Kasimir-Bauer S. Stem cell and epithelial-mesenchymal transition markers are frequently overexpressed in circulating tumor cells of metastatic breast cancer patients. Breast Cancer Res. 2009;11(4):R46.PubMedGoogle Scholar
  118. 118.
    Gupta PB, Onder TT, Jiang G, Tao K, Kuperwasser C, Weinberg RA, et al. Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell. 2009;138(4):645–59.PubMedGoogle Scholar
  119. 119.
    Hoeflich KP, O’Brien C, Boyd Z, Cavet G, Guerrero S, Jung K, et al. In vivo antitumor activity of MEK and phosphatidylinositol 3-kinase inhibitors in basal-like breast cancer models. Clin Cancer Res. 2009;15(14):4649–64.PubMedGoogle Scholar
  120. 120.
    Cho KB, Cho MK, Lee WY, and Kang KW. Overexpression of c-myc induces epithelial mesenchymal transition in mammary epithelial cells. Cancer Lett. 2010;293(2):230–9.Google Scholar
  121. 121.
    Wendt MK, Schiemann WP. Therapeutic targeting of the focal adhesion complex prevents oncogenic TGF-beta signaling and metastasis. Breast Cancer Res. 2009;11(5):R68.PubMedGoogle Scholar
  122. 122.
    Funasaka T, Hogan V, Raz A. Phosphoglucose isomerase/autocrine motility factor mediates epithelial and mesenchymal phenotype conversions in breast cancer. Cancer Res. 2009;69(13):5349–56.PubMedGoogle Scholar
  123. 123.
    Arima Y, Inoue Y, Shibata T, Hayashi H, Nagano O, Saya H, et al. Rb depletion results in deregulation of E-cadherin and induction of cellular phenotypic changes that are characteristic of the epithelial-to-mesenchymal transition. Cancer Res. 2008;68(13):5104–12.PubMedGoogle Scholar
  124. 124.
    Tumbarello DA, Turner CE. Hic-5 contributes to epithelial-mesenchymal transformation through a RhoA/ROCK-dependent pathway. J Cell Physiol. 2007;211(3):736–47.PubMedGoogle Scholar
  125. 125.
    Wu X, Chen H, Parker B, Rubin E, Zhu T, Lee JS, et al. HOXB7, a homeodomain protein, is overexpressed in breast cancer and confers epithelial-mesenchymal transition. Cancer Res. 2006;66(19):9527–34.PubMedGoogle Scholar
  126. 126.
    Chua HL, Bhat-Nakshatri P, Clare SE, Morimiya A, Badve S, Nakshatri H. NF-kappaB represses E-cadherin expression and enhances epithelial to mesenchymal transition of mammary epithelial cells: potential involvement of ZEB-1 and ZEB-2. Oncogene. 2007;26(5):711–24.PubMedGoogle Scholar
  127. 127.
    Bundy L, Wells S, Sealy L. C/EBPbeta-2 confers EGF-independent growth and disrupts the normal acinar architecture of human mammary epithelial cells. Mol Cancer. 2005;4:43.PubMedGoogle Scholar
  128. 128.
    Tanaka H, Shirkoohi R, Nakagawa K, Qiao H, Fujita H, Okada F, et al. siRNA gelsolin knockdown induces epithelial-mesenchymal transition with a cadherin switch in human mammary epithelial cells. Int J Cancer. 2006;118(7):1680–91.PubMedGoogle Scholar
  129. 129.
    Guan F, Handa K, Hakomori SI. Specific glycosphingolipids mediate epithelial-to-mesenchymal transition of human and mouse epithelial cell lines. Proc Natl Acad Sci U S A. 2009;106(18):7461–6.PubMedGoogle Scholar
  130. 130.
    Cheng GZ, Chan J, Wang Q, Zhang W, Sun CD, Wang LH. Twist transcriptionally up-regulates AKT2 in breast cancer cells leading to increased migration, invasion, and resistance to paclitaxel. Cancer Res. 2007;67(5):1979–87.PubMedGoogle Scholar
  131. 131.
    Hiscox S, Jiang WG, Obermeier K, Taylor K, Morgan L, Burmi R, et al. Tamoxifen resistance in MCF7 cells promotes EMT-like behaviour and involves modulation of beta-catenin phosphorylation. Int J Cancer. 2006;118(2):290–301.PubMedGoogle Scholar
  132. 132.
    Planas-Silva MD, Waltz PK. Estrogen promotes reversible epithelial-to-mesenchymal-like transition and collective motility in MCF-7 breast cancer cells. J Steroid Biochem Mol Biol. 2007;104(1–2):11–21.PubMedGoogle Scholar
  133. 133.
    Shtutman M, Levina E, Ohouo P, Baig M, Roninson IB. Cell adhesion molecule L1 disrupts E-cadherin-containing adherens junctions and increases scattering and motility of MCF7 breast carcinoma cells. Cancer Res. 2006;66(23):11370–80.PubMedGoogle Scholar
  134. 134.
    Micalizzi DS, Christensen KL, Jedlicka P, Coletta RD, Baron AE, Harrell JC, et al. The Six1 homeoprotein induces human mammary carcinoma cells to undergo epithelial-mesenchymal transition and metastasis in mice through increasing TGF-beta signaling. J Clin Invest. 2009;119(9):2678–90.PubMedGoogle Scholar
  135. 135.
    Allington TM, Galliher-Beckley AJ, Schiemann WP. Activated Abl kinase inhibits oncogenic transforming growth factor-beta signaling and tumorigenesis in mammary tumors. FASEB J. 2009;23(12):4231–43.PubMedGoogle Scholar
  136. 136.
    Neil JR, Johnson KM, Nemenoff RA, Schiemann WP. Cox-2 inactivates Smad signaling and enhances EMT stimulated by TGF-beta through a PGE2-dependent mechanisms. Carcinogenesis. 2008;29(11):2227–35.PubMedGoogle Scholar
  137. 137.
    Lee YH, Albig AR, Regner M, Schiemann BJ, Schiemann WP. Fibulin-5 initiates epithelial-mesenchymal transition (EMT) and enhances EMT induced by TGF-beta in mammary epithelial cells via a MMP-dependent mechanism. Carcinogenesis. 2008;29(12):2243–51.PubMedGoogle Scholar
  138. 138.
    Galliher AJ, Schiemann WP. Beta3 integrin and Src facilitate transforming growth factor-beta mediated induction of epithelial-mesenchymal transition in mammary epithelial cells. Breast Cancer Res. 2006;8(4):R42.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Tony Blick
    • 1
  • Honor Hugo
    • 1
  • Edwin Widodo
    • 2
    • 3
  • Mark Waltham
    • 1
    • 2
  • Cletus Pinto
    • 1
    • 2
  • Sendurai A. Mani
    • 4
  • Robert A. Weinberg
    • 5
  • Richard M. Neve
    • 6
    • 7
  • Marc E. Lenburg
    • 7
    • 8
  • Erik W. Thompson
    • 1
    • 2
    Email author
  1. 1.Invasion and Metastasis UnitSt. Vincent’s InstituteMelbourneAustralia
  2. 2.Department of SurgerySt. Vincent’s Hospital, University of MelbourneFitzroyAustralia
  3. 3.Faculty of MedicineBrawijaya UniversityEast JavaIndonesia
  4. 4.Department of Molecular Pathology, Unit 951The University of Texas M. D. Anderson Cancer CenterHoustonUSA
  5. 5.Whitehead Institute for Biomedical Research, 9 Cambridge Center, and Department of BiologyMassachusetts Institute of TechnologyCambridgeUSA
  6. 6.Molecular Biology DepartmentGenentech IncSouth San FranciscoUSA
  7. 7.Life Sciences DivisionLawrence Berkeley National LaboratoryBerkeleyUSA
  8. 8.Department of Pathology and Laboratory MedicineBoston University School of MedicineBostonUSA

Personalised recommendations