Cancer Microenvironment

, Volume 5, Issue 1, pp 19–28 | Cite as

Partial Mesenchymal to Epithelial Reverting Transition in Breast and Prostate Cancer Metastases

  • Yvonne Chao
  • Qian Wu
  • Marie Acquafondata
  • Rajiv Dhir
  • Alan WellsEmail author
Original Paper


Epithelial to mesenchymal transition (EMT) is an oft-studied mechanism for the initiation of metastasis. We have recently shown that once cancer cells disseminate to a secondary organ, a mesenchymal to epithelial reverting transition (MErT) may occur, which we postulate is to enable metastatic colonization. Despite a wealth of in vitro and in vivo studies, evidence supportive of MErT in human specimens is rare and difficult to document because clinically detectable metastases are typically past the micrometastatic stage at which this transition is most likely evident. We obtained paired primary and metastatic tumors from breast and prostate cancer patients and evaluated expression of various epithelial and mesenchymal markers by immunohistochemistry. The metastases exhibited increased expression of membranous E-cadherin compared to primary tumors, consistent with EMT at the primary site and MErT at the metastatic site. However, the re-emergence of the epithelial phenotype was only partial or incomplete. Expression of epithelial markers connexins 26 and/or 43 was also increased on the majority of metastases, particularly those to the brain. Despite the upregulation of epithelial markers in metastases, expression of mesenchymal markers vimentin and FSP1 was mostly unchanged. We also examined prostate carcinoma metastases of varied sizes and found that while E-cadherin expression was increased compared to the primary lesion, the expression inversely correlated with size of the metastasis. This not only suggests that a second EMT may occur in the ectopic site for tumor growth or to seed further metastases, but also provides a basis for the failure to discern epithelial phenotypes in clinically examined macrometastases. In summary, we report increased expression of epithelial markers and persistence of mesenchymal markers consistent with a partial MErT that readily allows for a second EMT at the metastatic site. Our results suggest that cancer cells continue to display phenotypic plasticity beyond the EMT that initiates metastasis.


Mesenchymal-to-Epithelial transition E-cadherin Differentiation Connexin 



These studies were supported by a Merit Award from the Veterans Administration and a predoctoral fellowship from the DoD CDMRP in Breast Cancer.


  1. 1.
    Wells A, Chao YL, Grahovac J, Wu Q, Lauffenburger DA (2011) Epithelial and mesenchymal phenotypic switchings modulate cell motility in metastasis. Front Biosci 16:815–837PubMedCrossRefGoogle Scholar
  2. 2.
    Condeelis JS, Wyckoff J, Segall JE (2000) Imaging of cancer invasion and metastasis using green fluorescent protein. Eur J Cancer 36:1671–1680PubMedCrossRefGoogle Scholar
  3. 3.
    Tarin D, Thompson EW, Newgreen DF (2005) The fallacy of epithelial mesenchymal transition in neoplasia. Cancer Res 65:5996–6000, discussion −1PubMedCrossRefGoogle Scholar
  4. 4.
    Wells A, Yates C, Shepard CR (2008) E-cadherin as an indicator of mesenchymal to epithelial reverting transitions during the metastatic seeding of disseminated carcinomas. Clin Exp Metastasis 25:621–628PubMedCrossRefGoogle Scholar
  5. 5.
    Thiery JP (2002) Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer 2:442–454PubMedCrossRefGoogle Scholar
  6. 6.
    Hugo H, Ackland ML, Blick T, Lawrence MG, Clements JA, Williams ED, Thompson EW (2007) Epithelial–mesenchymal and mesenchymal–epithelial transitions in carcinoma progression. J Cell Physiol 213:374–383PubMedCrossRefGoogle Scholar
  7. 7.
    Chaffer CL, Brennan JP, Slavin JL, Blick T, Thompson EW, Williams ED (2006) Mesenchymal-to-epithelial transition facilitates bladder cancer metastasis: role of fibroblast growth factor receptor-2. Cancer Res 66:11271–11278PubMedCrossRefGoogle Scholar
  8. 8.
    Chao YL, Shepard CR, Wells A (2010) Breast carcinoma cells re-express E-cadherin during mesenchymal to epithelial reverting transition. Mol Cancer 9:179PubMedCrossRefGoogle Scholar
  9. 9.
    Yates CC, Shepard CR, Stolz DB, Wells A (2007) Co-culturing human prostate carcinoma cells with hepatocytes leads to increased expression of E-cadherin. Br J Cancer 96:1246–1252PubMedCrossRefGoogle Scholar
  10. 10.
    Kowalski PJ, Rubin MA, Kleer CG (2003) E-cadherin expression in primary carcinomas of the breast and its distant metastases. Breast Cancer Res 5:R217–R222PubMedCrossRefGoogle Scholar
  11. 11.
    Bukholm IK, Nesland JM, Borresen-Dale AL (2000) Re-expression of E-cadherin, alpha-catenin and beta-catenin, but not of gamma-catenin, in metastatic tissue from breast cancer patients [seecomments]. J Pathol 190:15–19PubMedCrossRefGoogle Scholar
  12. 12.
    Wong AS, Gumbiner BM (2003) Adhesion-independent mechanism for suppression of tumor cell invasion by E-cadherin. J Cell Biol 161:1191–1203PubMedCrossRefGoogle Scholar
  13. 13.
    Brabletz T, Jung A, Reu S, Porzner M, Hlubek F, Kunz-Schughart LA, Knuechel R, Kirchner T (2001) Variable beta-catenin expression in colorectal cancers indicates tumor progression driven by the tumor environment. Proc Natl Acad Sci USA 98:10356–10361PubMedCrossRefGoogle Scholar
  14. 14.
    Monaghan P, Clarke C, Perusinghe NP, Moss DW, Chen XY, Evans WH (1996) Gap junction distribution and connexin expression in human breast. Exp Cell Res 223:29–38PubMedCrossRefGoogle Scholar
  15. 15.
    Li Z, Zhou Z, Welch DR, Donahue HJ (2008) Expressing connexin 43 in breast cancer cells reduces their metastasis to lungs. Clin Exp Metastasis 25:893–901PubMedCrossRefGoogle Scholar
  16. 16.
    McLachlan E, Shao Q, Wang HL, Langlois S, Laird DW (2006) Connexins act as tumor suppressors in three-dimensional mammary cell organoids by regulating differentiation and angiogenesis. Cancer Res 66:9886–9894PubMedCrossRefGoogle Scholar
  17. 17.
    Kanczuga-Koda L, Sulkowski S, Koda M, Sulkowska M (2005) Alterations in connexin26 expression during colorectal carcinogenesis. Oncology 68:217–222PubMedCrossRefGoogle Scholar
  18. 18.
    Kanczuga-Koda L, Sulkowski S, Lenczewski A, Koda M, Wincewicz A, Baltaziak M, Sulkowska M (2006) Increased expression of connexins 26 and 43 in lymph node metastases of breast cancer. J Clin Pathol 59:429–433PubMedCrossRefGoogle Scholar
  19. 19.
    Kanczuga-Koda L, Sulkowska M, Koda M, Rutkowski R, Sulkowski S (2007) Increased expression of gap junction protein–connexin 32 in lymph node metastases of human ductal breast cancer. Folia Histochem Cytobiol 45(Suppl 1):S175–S180PubMedGoogle Scholar
  20. 20.
    Trimboli AJ, Fukino K, de Bruin A, Wei G, Shen L, Tanner SM, Creasap N, Rosol TJ, Robinson ML, Eng C, Ostrowski MC, Leone G (2008) Direct evidence for epithelial-mesenchymal transitions in breast cancer. Cancer Res 68:937–945PubMedCrossRefGoogle Scholar
  21. 21.
    Okada H, Danoff TM, Kalluri R, Neilson EG (1997) Early role of Fsp1 in epithelial-mesenchymal transformation. Am J Physiol 273:F563–F574PubMedGoogle Scholar
  22. 22.
    Mendez MG, Kojima SI, Goldman RD (2010) Vimentin induces changes in cell shape, motility, and adhesion during the epithelial to mesenchymal transition. Faseb JGoogle Scholar
  23. 23.
    Vuoriluoto K, Haugen H, Kiviluoto S, Mpindi JP, Nevo J, Gjerdrum C, Tiron C, Lorens JB, Ivaska J (2010) Vimentin regulates EMT induction by Slug and oncogenic H-Ras and migration by governing Axl expression in breast cancer. OncogeneGoogle Scholar
  24. 24.
    Stoletov K, Kato H, Zardouzian E, Kelber J, Yang J, Shattil S, Klemke R (2010) Visualizing extravasation dynamics of metastatic tumor cells. J Cell Sci 123:2332–2341PubMedCrossRefGoogle Scholar
  25. 25.
    Yates C, Shepard CR, Papworth G, Dash A, Beer Stolz D, Tannenbaum S, Griffith L, Wells A (2007) Novel three-dimensional organotypic liver bioreactor to directly visualize early events in metastatic progression. Adv Cancer Res 97:225–246PubMedCrossRefGoogle Scholar
  26. 26.
    Giampieri S, Pinner S, Sahai E (2010) Intravital imaging illuminates transforming growth factor beta signaling switches during metastasis. Cancer Res 70:3435–3439PubMedCrossRefGoogle Scholar
  27. 27.
    Glinskii OV, Huxley VH, Glinsky GV, Pienta KJ, Raz A, Glinsky VV (2005) Mechanical entrapment is insufficient and intercellular adhesion is essential for metastatic cell arrest in distant organs. Neoplasia 7:522–527PubMedCrossRefGoogle Scholar
  28. 28.
    Pontes-Junior J, Reis ST, Dall’Oglio M, Neves de Oliveira LC, Cury J, Carvalho PA, Ribeiro-Filho LA, Moreira Leite KR, Srougi M (2009) Evaluation of the expression of integrins and cell adhesion molecules through tissue microarray in lymph node metastases of prostate cancer. J Carcinog 8:3PubMedCrossRefGoogle Scholar
  29. 29.
    Huang CF, Lira C, Chu K, Bilen MA, Lee YC, Ye X, Kim S, Ortiz A, Wu F, Logothetis C, Yu-Lee LY, Lin SH (2010) Cadherin-11 increases migration and invasion of prostate cancer cells and enhances their interaction with osteoblasts. Cancer Res 70:4580–4589PubMedCrossRefGoogle Scholar
  30. 30.
    Li Z, Zhou Z, Donahue HJ (2008) Alterations in Cx43 and OB-cadherin affect breast cancer cell metastatic potential. Clin Exp Metastasis 25:265–272PubMedCrossRefGoogle Scholar
  31. 31.
    Fidler IJ, Balasubramanian K, Lin Q, Kim SW, Kim SJ (2010) The brain microenvironment and cancer metastasis. Mol Cells 30:93–98PubMedCrossRefGoogle Scholar
  32. 32.
    Langley RR, Fan D, Guo L, Zhang C, Lin Q, Brantley EC, McCarty JH, Fidler IJ (2009) Generation of an immortalized astrocyte cell line from H-2Kb-tsA58 mice to study the role of astrocytes in brain metastasis. Int J Oncol 35:665–672PubMedCrossRefGoogle Scholar
  33. 33.
    Lin Q, Balasubramanian K, Fan D, Kim SJ, Guo L, Wang H, Bar-Eli M, Aldape KD, Fidler IJ (2010) Reactive astrocytes protect melanoma cells from chemotherapy by sequestering intracellular calcium through gap junction communication channels. Neoplasia 12:748–754PubMedGoogle Scholar
  34. 34.
    Christofori G (2006) New signals from the invasive front. Nature 441:444–450PubMedCrossRefGoogle Scholar
  35. 35.
    Friedl P, Gilmour D (2009) Collective cell migration in morphogenesis, regeneration and cancer. Nat Rev Mol Cell Biol 10:445–457PubMedCrossRefGoogle Scholar
  36. 36.
    Schmidmaier R, Baumann P (2008) ANTI-ADHESION evolves to a promising therapeutic concept in oncology. Curr Med Chem 15:978–990PubMedCrossRefGoogle Scholar
  37. 37.
    Gout S, Tremblay PL, Huot J (2008) Selectins and selectin ligands in extravasation of cancer cells and organ selectivity of metastasis. Clin Exp Metastasis 25:335–344PubMedCrossRefGoogle Scholar
  38. 38.
    Christiansen JJ, Rajasekaran AK (2006) Reassessing epithelial to mesenchymal transition as a prerequisite for carcinoma invasion and metastasis. Cancer Res 66:8319–8326PubMedCrossRefGoogle Scholar
  39. 39.
    Tarin D (2011) Cell and tissue interactions in carcinogenesis and metastasis and their clinical significance. Semin Cancer Biol 21:72–82PubMedCrossRefGoogle Scholar
  40. 40.
    Viadana E, Bross ID, Pickren JW (1973) An autopsy study of some routes of dissemination of cancer of the breast. Br J Cancer 27:336–340PubMedCrossRefGoogle Scholar
  41. 41.
    Alterman AL, Fornabaio DM, Stackpole CW (1985) Metastatic dissemination of B16 melanoma: pattern and sequence of metastasis. J Natl Cancer Inst 75:691–702PubMedGoogle Scholar
  42. 42.
    Stackpole CW (1990) Intrapulmonary spread of established B16 melanoma lung metastases and lung colonies. Invasion Metastasis 10:267–280PubMedGoogle Scholar
  43. 43.
    Hoover HC Jr, Ketcham AS (1975) Metastasis of metastases. Am J Surg 130:405–411PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Yvonne Chao
    • 1
  • Qian Wu
    • 1
  • Marie Acquafondata
    • 1
  • Rajiv Dhir
    • 1
  • Alan Wells
    • 1
    • 2
    Email author
  1. 1.Department of PathologyPittsburgh VAMC and University of PittsburghPittsburghUSA
  2. 2.School of MedicineUniversity of PittsburghPittsburghUSA

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