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Enrichment, Isolation and Molecular Characterization of EpCAM-Negative Circulating Tumor Cells

  • Rita Lampignano
  • Helen Schneck
  • Martin Neumann
  • Tanja Fehm
  • Hans NeubauerEmail author
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 994)

Abstract

The presence of EpCAM-positive circulating tumor cells (CTCs) in the peripheral blood is associated with poor clinical outcomes in breast, colorectal and prostate cancer, as well as the prognosis of other tumor types. In addition, recent studies have suggested that the presence of CTCs undergoing epithelial-to-mesenchymal transition and, as such, may exhibit reduced or no expression of epithelial proteins e.g. EpCAM, might be related to disease progression in metastatic breast cancer (MBC) patients. Analyzing the neoplastic nature of this EpCAM-low/negative (EpCAM-neg) subpopulation remains an open issue as the current standard detection methods for CTCs are not efficient at identifying this subpopulation of cells. The possible association of EpCAM-neg CTCs with EpCAM-positive (EpCAM-pos) CTCs and role in the clinicopathological features and prognosis of MBC patients has still to be demonstrated. Several technologies have been developed and are currently being tested for the identification and the downstream analyses of EpCAM-pos CTCs. These technologies can be adapted and implemented into workflows to isolate and investigate EpCAM-neg cells to understand their biology and clinical relevance. This chapter will endeavour to explain the rationale behind the identification and analyses of all CTC subgroups, as well as to review the current strategies employed to enrich, isolate and characterize EpCAM-negative CTCs. Finally, the latest findings in the field will briefly be discussed with regard to their clinical relevance.

Keywords

Single cell isolation Molecular analysis EPCAM-negative 

References

  1. Aceto N (2014) Circulating tumor cell clusters are oligoclonal precursors of breast cancer metastasis. Cell 158:1110–1122PubMedPubMedCentralCrossRefGoogle Scholar
  2. Adams D, Makarova O, Zhu P et al (2011) Abstract 2369: isolation of circulating tumor cells by size exclusion using lithography fabricated precision microfilters. Cancer Res 71(8 Suppl):2369–2369CrossRefGoogle Scholar
  3. Aktas B (2009) Stem cell and epithelial-mesenchymal transition markers are frequently overexpressed in circulating tumor cells of metastatic breast cancer patients. Breast Cancer Res 11:R46PubMedPubMedCentralCrossRefGoogle Scholar
  4. Albuquerque A, Kaul S, Breier G et al (2012) Multimarker analysis of circulating tumor cells in peripheral blood of metastatic breast cancer patients: a step forward in personalized medicine. Breast Care 7:7–12PubMedPubMedCentralCrossRefGoogle Scholar
  5. Alix-Panabières C, Pantel K (2013) Technologies for detection of circulating tumor cells: facts and vision. Lab Chip 14(1):57–62PubMedCrossRefGoogle Scholar
  6. Allan AL, Keeney M (2010) Circulating tumor cell analysis: technical and statistical considerations for application to the clinic. J Oncol 2010:426218PubMedCrossRefGoogle Scholar
  7. Allard WJ, Matera J, Miller MC et al (2004) Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin Cancer Res 10(20):6897–6904PubMedCrossRefGoogle Scholar
  8. Balic M, Dandachi N, Hofmann G et al (2005) Comparison of two methods for enumerating circulating tumor cells in carcinoma patients. Cytometry B Clin Cytom 68B(1):25–30CrossRefGoogle Scholar
  9. Balzar M, Winter MJ, de Boer CJ, Litvinov SV (1999) The biology of the 17-1A antigen (Ep-CAM). J Mol Med Berl Ger 77(10):699–712CrossRefGoogle Scholar
  10. Barradas AMC, Terstappen LWMM (2013) Towards the biological understanding of CTC: capture technologies, definitions and potential to create metastasis. Cancers 5(4):1619–1642Google Scholar
  11. Bidard FC (2014) Clinical validity of circulating tumour cells in patients with metastatic breast cancer: a pooled analysis of individual patient data. Lancet Oncol 15:406–414PubMedCrossRefGoogle Scholar
  12. Bitting RL, Boominathan R, Rao C et al (2013) Development of a method to isolate circulating tumor cells using mesenchymal-based capture. Methods San Diego Calif 64(2):129–136CrossRefGoogle Scholar
  13. Bolognesi C, Forcato C, Buson G et al (2016) Digital sorting of pure cell populations enables unambiguous genetic analysis of heterogeneous formalin-Fixed paraffin-embedded tumors by next generation sequencing. Sci Rep 6:20944PubMedPubMedCentralCrossRefGoogle Scholar
  14. Bozzetti C, Quaini F, Squadrilli A et al (2015) Isolation and characterization of circulating tumor cells in squamous cell carcinoma of the lung using a non-EpCAM-based capture method. PLoS One 10:e0142891PubMedPubMedCentralCrossRefGoogle Scholar
  15. Bulfoni M, Gerratana L, Del Ben F et al (2016) In patients with metastatic breast cancer the identification of circulating tumor cells in epithelial-to-mesenchymal transition is associated with a poor prognosis. Breast Cancer Res 18:1–15CrossRefGoogle Scholar
  16. Chang MC (2016) Clinical significance of circulating tumor microemboli as a prognostic marker in patients with pancreatic ductal adenocarcinoma. Clin Chem 62(3):505–513PubMedCrossRefGoogle Scholar
  17. Chen C-L, Mahalingam D, Osmulski P et al (2013) Single-cell analysis of circulating tumor cells identifies cumulative expression patterns of EMT-related genes in metastatic prostate cancer. Prostate 73(8):813–826PubMedCrossRefGoogle Scholar
  18. Cho EH, Wendel M, Luttgen M et al (2012) Characterization of circulating tumor cell aggregates identified in patients with epithelial tumors. Phys Biol 9(1):16001CrossRefGoogle Scholar
  19. Chudziak J, Burt DJ, Mohan S et al (2016) Clinical evaluation of a novel microfluidic device for epitope-independent enrichment of circulating tumour cells in patients with small cell lung cancer. Analyst 141(2):669–678PubMedCrossRefGoogle Scholar
  20. Chung Y-K, Reboud J, Lee KC et al (2011) An electrical biosensor for the detection of circulating tumor cells. Biosens Bioelectron 26(5):2520–2526PubMedCrossRefGoogle Scholar
  21. Cohen SJ, Punt CJ, Iannotti N et al (2008) Relationship of circulating tumour cells to tumour response, progression-free survival, and overall survival in patients with metastatic colorectal cancer. J Clin Oncol 26(19):3213–3221PubMedCrossRefGoogle Scholar
  22. Coumans FAW, van Dalum G, Beck M et al (2013) Filter Characteristics Influencing Circulating Tumor Cell Enrichment from Whole Blood. PLoS One 8(4):e61770PubMedPubMedCentralCrossRefGoogle Scholar
  23. Cristofanilli M, Budd GT, Ellis MJ et al (2004) Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med 351:781–791PubMedCrossRefGoogle Scholar
  24. de Bono JS, Scher HI, Montgomery RB et al (2008) Circulating tumour cells predict survival benefit from treatment in metastatic castration-resistant prostate cancer. Clin Cancer Res Off J Am Assoc Cancer Res 14(19):6302–6309CrossRefGoogle Scholar
  25. De Giorgi V, Pinzani P, Salvianti F et al (2010) Application of a filtration- and isolation-by-size technique for the detection of circulating tumor cells in cutaneous melanoma. J Invest Dermatol 130(10):2440–2447PubMedCrossRefGoogle Scholar
  26. de Wit S, Dalum G van, Lenferink ATM et al (2015) The detection of EpCAM+ and EpCAM– circulating tumour cells. Sci Rep 5:12270.Google Scholar
  27. Desitter I, Guerrouahen BS, Benali-Furet N et al (2011) A new device for rapid isolation by size and characterization of rare circulating tumor cells. Anticancer Res 31(2):427–441PubMedGoogle Scholar
  28. Driemel C, Kremling H, Schumacher S et al (2014) Context-dependent adaption of EpCAM expression in early systemic esophageal cancer. Oncogene 33(41):4904–4915PubMedCrossRefGoogle Scholar
  29. Esmaeilsabzali H, Beischlag TV, Cox ME et al (2013) Detection and isolation of circulating tumor cells: principles and methods. Biotechnol Adv 31(7):1063–1084PubMedCrossRefGoogle Scholar
  30. Fabbri F, Carloni S, Zoli W et al (2013) Detection and recovery of circulating colon cancer cells using a dielectrophoresis-based device: KRAS mutation status in pure CTCs. Cancer Lett 335(1):225–231PubMedCrossRefGoogle Scholar
  31. Farace F, Massard C, Vimond N et al (2011) A direct comparison of Cell Search and ISET for circulating tumour-cell detection in patients with metastatic carcinomas. Br J Cancer 105(6):847–853PubMedPubMedCentralCrossRefGoogle Scholar
  32. Flieger D, Hoff AS, Sauerbruch T et al (2001) Influence of cytokines, monoclonal antibodies and chemotherapeutic drugs on epithelial cell adhesion molecule (EpCAM) and Lewis Y antigen expression. Clin Exp Immunol 123(1):9–14PubMedPubMedCentralCrossRefGoogle Scholar
  33. Gabriel MT, Calleja LR, Chalopin A et al (2016) Circulating tumor cells: a review of non–EpCAM-based approaches for cell enrichment and isolation. Clin Chem 62(4):571–581PubMedCrossRefGoogle Scholar
  34. Galletti G, Sung MS, Vahdat LT et al (2014) Isolation of breast cancer and gastric cancer circulating tumor cells by use of an anti HER2-based microfluidic device. Lab Chip 14(1):147–156PubMedCrossRefGoogle Scholar
  35. Gires O, Kieu C, Fix P et al (2001) Tumor necrosis factor alpha negatively regulates the expression of the carcinoma-associated antigen epithelial cell adhesion molecule. Cancer 92(3):620–628PubMedCrossRefGoogle Scholar
  36. Gires O, Stoecklein NH (2014) Dynamic EpCAM expression on circulating and disseminating tumour cells: causes and consequences. Cell Mol Life Sci 71(22):4393–4402PubMedCrossRefGoogle Scholar
  37. Gorges TM, Tinhofer I, Drosch M et al (2012) Circulating tumour cells escape from EpCAM-based detection due to epithelial-to-mesenchymal transition. BMC Cancer 12:178PubMedPubMedCentralCrossRefGoogle Scholar
  38. He W, Kularatne SA, Kalli KR et al (2008) Quantitation of circulating tumor cells in blood samples from ovarian and prostate cancer patients using tumor-specific fluorescent ligands. Int J Cancer 123(8):1968–1973PubMedPubMedCentralCrossRefGoogle Scholar
  39. Herlyn D, Herlyn M, Steplewski Z et al (1979) Monoclonal antibodies in cell-mediated cytotoxicity against human melanoma and colorectal carcinoma. Eur J Immunol 9(8):657–659PubMedCrossRefGoogle Scholar
  40. Hiltermann TJN, Pore MM, van den Berg A et al (2012) Circulating tumour cells in small-cell lung cancer: a predictive and prognostic factor. Ann Oncol 23(11):2937–2942PubMedCrossRefGoogle Scholar
  41. Hofman V, Ilie MI, Long E et al (2011) Detection of circulating tumor cells as a prognostic factor in patients undergoing radical surgery for non-small-cell lung carcinoma: comparison of the efficacy of the CellSearch Assay™ and the isolation by size of epithelial tumor cell method. Int J Cancer 129(7):1651–1660PubMedCrossRefGoogle Scholar
  42. Hoshino K, Huang Y-Y, Lane N et al (2011) Microchip-based immunomagnetic detection of circulating tumor cell. Lab Chip 11(20):3449–3457PubMedPubMedCentralCrossRefGoogle Scholar
  43. Hou HW, Li QS, Lee GYH et al (2008) Deformability study of breast cancer cells using microfluidics. Biomed Microdevices 11(3):557–564CrossRefGoogle Scholar
  44. Hou HW, Warkiani ME, Khoo BL et al (2013) Isolation and retrieval of circulating tumor cells using centrifugal forces. Sci Rep 3:1259PubMedPubMedCentralCrossRefGoogle Scholar
  45. Hur SC, Mach AJ, Di Carlo D (2011) High-throughput size-based rare cell enrichment using microscale vortices. Biomicrofluidics 5(2):022206PubMedCentralCrossRefGoogle Scholar
  46. Hvichia GE, Parveen Z, Wagner C et al (2016) A novel microfluidic platform for size and deformability based separation and the subsequent molecular characterization of viable circulating tumor cells. Int J Cancer 138(12):2894–2904PubMedPubMedCentralCrossRefGoogle Scholar
  47. Hyun K-A, Koo G-B, Han H et al (2016) Epithelial-to-mesenchymal transition leads to loss of EpCAM and different physical properties in circulating tumor cells from metastatic breast cancer. Oncotarget 7(17):24677–24687PubMedPubMedCentralGoogle Scholar
  48. Joosse SA, Gorges TM, Pantel K (2015) Biology, detection, and clinical implications of circulating tumor cells. EMBO Mol Med 7(1):1–11PubMedCrossRefGoogle Scholar
  49. Kahn HJ, Presta A, Yang L-Y et al (2004) Enumeration of circulating tumor cells in the blood of breast cancer patients after filtration enrichment: correlation with disease stage. Breast Cancer Res Treat 86(3):237–247PubMedCrossRefGoogle Scholar
  50. Khoja L, Backen A, Sloane R et al (2012) A pilot study to explore circulating tumour cells in pancreatic cancer as a novel biomarker. Br J Cancer 106(3):508–516PubMedCrossRefGoogle Scholar
  51. Khoo BL, Warkiani ME, Tan DS-W et al (2014) Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells. PLoS One 9(7):e99409PubMedPubMedCentralCrossRefGoogle Scholar
  52. Kirby BJ, Jodari M, Loftus MS et al (2012) Functional characterization of circulating tumor cells with a prostate-cancer-specific microfluidic device. PLoS One 7(4):e35976PubMedPubMedCentralCrossRefGoogle Scholar
  53. Konigsberg R, Obermayr E, Bises G et al (2011) Detection of EpCAM positive and negative circulating tumor cells in metastatic breast cancer patients. Acta Oncol 50(5):700–710PubMedCrossRefGoogle Scholar
  54. Krebs MG, Sloane R, Priest L et al (2011) Evaluation and prognostic significance of circulating tumor cells in patients with non–small-cell lung cancer. J Clin Oncol 29(12):1556–1563Google Scholar
  55. Krebs MG, Hou J-M, Sloane R et al (2012) Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and -independent approaches. J Thorac Oncol 7(2):306–315PubMedCrossRefGoogle Scholar
  56. Lapin M, Tjensvoll K, Oltedal S et al (2016) MINDEC-an enhanced negative depletion strategy for circulating tumour cell enrichment. Sci Rep 6:28929PubMedPubMedCentralCrossRefGoogle Scholar
  57. Lecharpentier A, Vielh P, Perez-Moreno P et al (2011) Detection of circulating tumour cells with a hybrid (epithelial/mesenchymal) phenotype in patients with metastatic non-small cell lung cancer. Br J Cancer 105(9):1338–1341PubMedPubMedCentralCrossRefGoogle Scholar
  58. Leong FY, Li Q, Lim CT, Chiam K-H (2010) Modeling cell entry into a micro-channel. Biomech Model Mechanobiol 10(5):755–766PubMedCrossRefGoogle Scholar
  59. Lianidou ES, Mavroudis D, Georgoulias V (2013) Clinical challenges in the molecular characterization of circulating tumour cells in breast cancer. Br J Cancer 108:2426–2432PubMedPubMedCentralCrossRefGoogle Scholar
  60. Lin HK, Zheng S, Williams AJ et al (2010) Portable filter-based microdevice for detection and characterization of circulating tumor cells. Am Assoc Cancer Res 16(20):5011–5018Google Scholar
  61. Lin M, Chen J-F, Lu Y-T et al (2014) Nanostructure embedded microchips for detection, isolation, and characterization of circulating tumor cells. Acc Chem Res 47(10):2941–2950PubMedPubMedCentralCrossRefGoogle Scholar
  62. Liu Z (2011) Negative enrichment by immunomagnetic nanobeads for unbiased characterization of circulating tumor cells from peripheral blood of cancer patients. J Transl Med 9:70–70PubMedPubMedCentralCrossRefGoogle Scholar
  63. Lu Y (2015) Isolation and characterization of living circulating tumor cells in patients by immunomagnetic negative enrichment coupled with flow cytometry. Cancer 121(17):3036–3045PubMedCrossRefGoogle Scholar
  64. Lustberg MB, Balasubramanian P, Miller B et al (2014) Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients. Breast Cancer Res 16(2):R23PubMedPubMedCentralCrossRefGoogle Scholar
  65. Mach AJ, Kim JH, Arshi A et al (2011) Automated cellular sample preparation using a Centrifuge-on-a-Chip. Lab Chip 11(17):2827–2834PubMedCrossRefGoogle Scholar
  66. Maetzel D, Denzel S, Mack B et al (2009) Nuclear signalling by tumour-associated antigen EpCAM. Nat Cell Biol 11(2):162–171PubMedCrossRefGoogle Scholar
  67. Magbanua MJM, Park JW (2014) Advances in genomic characterization of circulating tumour cells. Cancer Metastasis Rev 33(2–3):757–769PubMedCrossRefGoogle Scholar
  68. Marrinucci D, Bethel K, Bruce RH et al (2007) Case study of the morphologic variation of circulating tumor cells. Hum Pathol 38(3):514–519PubMedCrossRefGoogle Scholar
  69. Marrinucci D, Bethel K, Kolatkar A et al (2012) Fluid biopsy in patients with metastatic prostate, pancreatic and breast cancers. Phys Biol 9(1):16003CrossRefGoogle Scholar
  70. Mego M (2012) Prognostic value of EMT-circulating tumor cells in metastatic breast cancer patients undergoing high-dose chemotherapy with autologous hematopoietic stem cell transplantation. J Cancer 3:369–380PubMedPubMedCentralCrossRefGoogle Scholar
  71. Mego M, De Giorgi U, Dawood S et al (2011) Characterization of metastatic breast cancer patients with nondetectable circulating tumor cells. Int J Cancer 129(2):417–423PubMedCrossRefGoogle Scholar
  72. Mikolajczyk SD, Millar LS, Tsinberg P et al (2011) Detection of EpCAM-negative and cytokeratin-negative circulating tumor cells in peripheral blood. J Oncol 2011:252361PubMedPubMedCentralCrossRefGoogle Scholar
  73. Morris KL, Tugwood JD, Khoja L et al (2014) Circulating biomarkers in hepatocellular carcinoma. Cancer Chemother Pharmacol 74(2):323–332PubMedCrossRefGoogle Scholar
  74. Mostert B, Kraan J, Bolt-de Vries J et al (2011) Detection of circulating tumor cells in breast cancer may improve through enrichment with anti-CD146. Breast Cancer Res Treat 127(1):33–41PubMedCrossRefGoogle Scholar
  75. Mostert B, Kraan J, Sieuwerts AM et al (2012) CD49f-based selection of circulating tumor cells (CTCs) improves detection across breast cancer subtypes. Cancer Lett 319(1):49–55PubMedCrossRefGoogle Scholar
  76. Müller V, Stahmann N, Riethdorf S et al (2005) Circulating tumor cells in breast cancer: correlation to bone marrow micrometastases, heterogeneous response to systemic therapy and low proliferative activity. Am Assoc Cancer Res 11(10):3678–3685Google Scholar
  77. Neumann MHD, Schneck H, Decker Y et al (2016) Isolation and characterization of circulating tumor cells using a novel workflow combining the CellSearch® system and the CellCelector™. Biotechnol Prog.Google Scholar
  78. Neves RPL, Raba K, Schmidt O et al (2014) Genomic high-resolution profiling of single CKpos/CD45neg flow-sorting purified circulating tumor cells from patients with metastatic breast cancer. Clin Chem 60(10):1290–1297PubMedGoogle Scholar
  79. Ntouroupi TG, Ashraf SQ, McGregor SB et al (2008) Detection of circulating tumour cells in peripheral blood with an automated scanning fluorescence microscope. Br J Cancer 99(5):789–795PubMedPubMedCentralCrossRefGoogle Scholar
  80. Pailler E, Adam J, Barthélémy A et al (2013) Detection of circulating tumor cells harboring a unique ALK rearrangement in ALK-positive non–small-cell lung cancer. J Clin Oncol 31(18):2273–2281PubMedCrossRefGoogle Scholar
  81. Pantel K, Alix-Panabières C (2010) Circulating tumour cells in cancer patients: challenges and perspectives. Trends Mol Med 16(9):398–406PubMedCrossRefGoogle Scholar
  82. Parkinson DR, Dracopoli N, Petty BG et al (2012) Considerations in the development of circulating tumor cell technology for clinical use. J Transl Med 10:138PubMedPubMedCentralCrossRefGoogle Scholar
  83. Patriarca C, Macchi RM, Marschner AK et al (2012) Epithelial cell adhesion molecule expression (CD326) in cancer: a short review. Cancer Treat Rev 38(1):68–75PubMedCrossRefGoogle Scholar
  84. Polzer B, Medoro G, Pasch S et al (2014) Molecular profiling of single circulating tumor cells with diagnostic intention. EMBO Mol Med 6(11):1371–1386PubMedPubMedCentralCrossRefGoogle Scholar
  85. Poruk KE (2016) Circulating tumor cell phenotype predicts recurrence and survival in pancreatic adenocarcinoma. Ann Surg 264(6):1073–1081PubMedPubMedCentralCrossRefGoogle Scholar
  86. Pösel C, Möller K, Fröhlich W et al (2012) Density gradient centrifugation compromises bone marrow mononuclear cell yield. PLoS One 7(12):e50293PubMedPubMedCentralCrossRefGoogle Scholar
  87. Pribluda A, de la Cruz CC, Jackson EL (2015) Intratumoral heterogeneity: from diversity comes resistance. Am Assoc Cancer Res 21(13):2916–2923Google Scholar
  88. Punnoose EA, Atwal SK, Spoerke JM et al (2010) Molecular biomarker analyses using circulating tumor cells. PLoS One 5(9):e12517PubMedPubMedCentralCrossRefGoogle Scholar
  89. Rao CG, Chianese D, Doyle GV et al (2005) Expression of epithelial cell adhesion molecule in carcinoma cells present in blood and primary and metastatic tumors. Int J Oncol 27(1):49–57PubMedGoogle Scholar
  90. Romsdahl MM, Valaitis J, McGrath RG et al (1965) Circulating tumor cells in patients with carcinoma: method and recent studies. JAMA 193(13):1087–1090PubMedCrossRefGoogle Scholar
  91. Rosenberg R, Gertler R, Friederichs J et al (2002) Comparison of two density gradient centrifugation systems for the enrichment of disseminated tumor cells in blood. Cytometry 49(4):150–158PubMedCrossRefGoogle Scholar
  92. Sarrió D, Rodriguez-Pinilla SM, Hardisson D et al (2008) Epithelial-mesenchymal transition in breast cancer relates to the basal-like phenotype. Cancer Res 68(4):989–997PubMedCrossRefGoogle Scholar
  93. Saucedo-Zeni N, Mewes S, Niestroj R et al (2012) A novel method for the in vivo isolation of circulating tumor cells from peripheral blood of cancer patients using a functionalized and structured medical wire. Int J Oncol 41(4):1241–1250PubMedPubMedCentralGoogle Scholar
  94. Schindlbeck C, Stellwagen J, Jeschke U et al (2008) Immunomagnetic enrichment of disseminated tumor cells in bone marrow and blood of breast cancer patients by the Thomsen-Friedenreich-Antigen. Clin Exp Metastasis 25(3):233–240PubMedCrossRefGoogle Scholar
  95. Schneck H, Gierke B, Uppenkamp F et al (2015) EpCAM-independent enrichment of circulating tumor cells in metastatic breast cancer. PLoS One 10(12):e0144535PubMedPubMedCentralCrossRefGoogle Scholar
  96. Seal SH (1959) Silicone flotation: a simple quantitative method for the isolation of free-floating cancer cells from the blood. Cancer 12(3):590–595PubMedCrossRefGoogle Scholar
  97. Seal SH (1964) A sieve for the isolation of cancer cells and other large cells from the blood. Cancer 17(5):637–642PubMedCrossRefGoogle Scholar
  98. Shaw Bagnall J, Byun S, Begum S et al (2015) Deformability of tumor cells versus blood cells. Sci Rep 5:18542PubMedPubMedCentralCrossRefGoogle Scholar
  99. Sieuwerts AM, Kraan J, Bolt J et al (2009) Anti-epithelial cell adhesion molecule antibodies and the detection of circulating normal-like breast tumor cells. J Natl Cancer Inst 101(1):61–66PubMedPubMedCentralCrossRefGoogle Scholar
  100. Smerage JB, Barlow WE, Hortobagyi GN et al (2014) Circulating tumor cells and response to chemotherapy in metastatic breast cancer: SWOG S0500. J Clin Oncol 32(31):3483–3489PubMedPubMedCentralCrossRefGoogle Scholar
  101. Sollier E, Go DE, Che J et al (2013) Size-selective collection of circulating tumor cells using Vortex technology. Lab Chip 14(1):63–77PubMedCrossRefGoogle Scholar
  102. Spizzo G, Gastl G, Obrist P et al (2007) Methylation status of the Ep-CAM promoter region in human breast cancer cell lines and breast cancer tissue. Cancer Lett 246(1–2):253–261PubMedCrossRefGoogle Scholar
  103. Stott SL, Hsu C-H, Tsukrov DI et al (2010) Isolation of circulating tumor cells using a microvortex-generating herringbone-chip. Proc Natl Acad Sci U S A 107(43):18392–18397PubMedPubMedCentralCrossRefGoogle Scholar
  104. Tai K-Y, Shiah S-G, Shieh Y-S et al (2007) DNA methylation and histone modification regulate silencing of epithelial cell adhesion molecule for tumor invasion and progression. Oncogene 26(27):3989–3997PubMedCrossRefGoogle Scholar
  105. Tan SJ, Yobas L, Lee GYH et al (2009) Microdevice for the isolation and enumeration of cancer cells from blood. Biomed Microdevices 11(4):883–892PubMedCrossRefGoogle Scholar
  106. Terstappen LW, Rao C, Gross S et al (1998) Flow cytometry–principles and feasibility in transfusion medicine. Enumeration of epithelial derived tumor cells in peripheral blood. Vox Sang 74(Suppl 2):269–274PubMedCrossRefGoogle Scholar
  107. Thiery JP (2002) Epithelial–mesenchymal transitions in tumour progression. Nat Rev Cancer 2(6):442–454PubMedCrossRefGoogle Scholar
  108. Thiery JP (2009) Epithelial-mesenchymal transitions in cancer onset and progression. Bull Acad Natl Med 193 (9):1969–1978 discussion 1978–9Google Scholar
  109. Tibbe AGJ, Miller MC, Terstappen LWMM (2007) Statistical considerations for enumeration of circulating tumor cells. Cytometry A 71A(3):154–162CrossRefGoogle Scholar
  110. Ueo H (2015) Circulating tumour cell-derived plastin3 is a novel marker for predicting long-term prognosis in patients with breast cancer. Br J Cancer 112:1519–1526PubMedPubMedCentralCrossRefGoogle Scholar
  111. Vishnoi M, Peddibhotla S, Yin W et al (2015) The isolation and characterization of CTC subsets related to breast cancer dormancy. Sci Rep 5:17533PubMedPubMedCentralCrossRefGoogle Scholar
  112. Vona G, Sabile A, Louha M et al (2000) Isolation by size of epithelial tumor cells. Am J Pathol 156(1):57–63PubMedPubMedCentralCrossRefGoogle Scholar
  113. Vona G, Estepa L, Béroud C et al (2004) Impact of cytomorphological detection of circulating tumor cells in patients with liver cancer. Hepatology 39(3):792–797PubMedCrossRefGoogle Scholar
  114. Wang S, Liu K, Liu J et al (2011) Highly efficient capture of circulating tumor cells by using nanostructured silicon substrates with integrated chaotic micromixers. Angew Chem Int Ed 50(13):3084–3088CrossRefGoogle Scholar
  115. Weitz J, Kienle P, Lacroix J et al (1998) Dissemination of tumor cells in patients undergoing surgery for colorectal cancer. Am Assoc Cancer Res 4(2):343–348Google Scholar
  116. Xu L, Mao X, Imrali A et al (2015) Optimization and evaluation of a novel size based circulating tumor cell isolation system. PLoS One 10(9):e0138032PubMedPubMedCentralCrossRefGoogle Scholar
  117. Yang L, Lang JC, Balasubramanian P et al (2009) Optimization of an enrichment process for circulating tumor cells from the blood of head and neck cancer patients through depletion of normal cells. Biotechnol Bioeng 102(2):521–534PubMedPubMedCentralCrossRefGoogle Scholar
  118. Yokobori T, Linuma H, Shimamura T et al (2013) Plastin3 is a novel marker for circulating tumor cells undergoing the epithelial-mesenchymal transition and is associated with colorectal cancer prognosis. Cancer Res 73:2059–2069Google Scholar
  119. Yu M, Bardia A, Wittner BS et al (2013) Circulating breast tumor cells exhibit dynamic changes in epithelial and mesenchymal composition. Science 339(6119):580–584PubMedPubMedCentralCrossRefGoogle Scholar
  120. Zhang L, Ridgway LD, Wetzel MD et al (2013) The identification and characterization of breast cancer CTCs competent for brain metastasis. Sci Transl Med 5(180):180ra48PubMedCrossRefGoogle Scholar
  121. Zheng S, Lin H, Liu J-Q et al (2007) Membrane microfilter device for selective capture, electrolysis and genomic analysis of human circulating tumor cells. J Chromatogr A 1162(2):154–161PubMedCrossRefGoogle Scholar
  122. Zheng S, Lin HK, Lu B et al (2011) 3D microfilter device for viable circulating tumor cell (CTC) enrichment from blood. Bimed Microdevices 13(1)Google Scholar

Copyright information

© Springer International Publishing AG 2017

Authors and Affiliations

  • Rita Lampignano
    • 1
  • Helen Schneck
    • 1
  • Martin Neumann
    • 1
  • Tanja Fehm
    • 1
  • Hans Neubauer
    • 2
    Email author
  1. 1.Department of Obstetrics and GynecologyUniversity Hospital and Medical Faculty of the Heinrich-Heine University DuesseldorfDuesseldorfGermany
  2. 2.Forschungslabore der Frauenklinik desUniversitätsklinikums Düsseldorf, Life Science CenterDüsseldorfGermany

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