Skip to main content
SpringerLink
Log in

Circulating Tumor Cells: Markers and Methodologies for Enrichment and Detection

  • Protocol
  • First Online:
Book cover Circulating Tumor Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1634))

Abstract

Cancer is a leading cause of disease worldwide; however, nowadays many points of its initiation processes are unknown. In this chapter, we are focusing on the role of liquid biopsies in cancer detection and progression. CTCs are one of the main components of liquid biopsies, they represent a subset of tumor cells that have acquired the ability to disseminate from the primary tumor and intravasate to the circulatory system. The greatest challenge in the detection of CTCs is their rarity in the blood. Human blood consists of white blood cells (5–10 × 106/mL), red blood cells (5–9 × 109/mL), and platelets (2.5–4 × 108/mL); very few CTCs will be present even in patients with known metastatic disease, with often less than one CTC per mL of blood. CTCs are found in frequencies on the order of 1–10 CTCs per mL of whole blood in patients with metastatic disease, and it is reduced in half for non-metastatic stages. Therefore, accurate methodologies for their capture and analysis are really important. The main aim of the present chapter is to describe different methodologies for CTCs capturing and analysis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 139.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 179.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 179.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Serrano MJ, Ortega FG, Alvarez-Cubero MJ, Nadal R, Sanchez-Rovira P, Salido M et al (2014) EMT and EGFR in CTCs cytokeratin negative non-metastatic breast cancer. Oncotarget 5(17):7486–7497

    Article  PubMed  PubMed Central  Google Scholar 

  2. Yu M, Stott S, Toner M, Maheswaran S, Haber DA (2011) Circulating tumor cells: approaches to isolation and characterization. J Cell Biol 192(3):373–382

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Lianidou ES, Markou A, Strati A (2015) The role of CTCs as tumor biomarkers. Adv Exp Med Biol 867:341–367

    Article  CAS  PubMed  Google Scholar 

  4. Satelli A, Brownlee Z, Mitra A, Meng QH, Li S (2015) Circulating tumor cell enumeration with a combination of epithelial cell adhesion molecule- and cell-surface vimentin-based methods for monitoring breast cancer therapeutic response. Clin Chem 61(1):259–266

    Article  CAS  PubMed  Google Scholar 

  5. Schneck H, Gierke B, Uppenkamp F, Behrens B, Niederacher D, Stoecklein NH et al (2015) EpCAM-independent enrichment of circulating tumor cells in metastatic breast cancer. PLoS One 10(12):e0144535

    Article  PubMed  PubMed Central  Google Scholar 

  6. Pantel K, Brakenhoff RH (2004) Dissecting the metastatic cascade. Nat Rev Cancer 4(6):448–456

    Article  CAS  PubMed  Google Scholar 

  7. Pierga JY, Bonneton C, Vincent-Salomon A, de Cremoux P, Nos C, Blin N et al (2004) Clinical significance of immunocytochemical detection of tumor cells using digital microscopy in peripheral blood and bone marrow of breast cancer patients. Clin Cancer Res 10(4):1392–1400

    Article  CAS  PubMed  Google Scholar 

  8. Templeton AJ, Diez-Gonzalez L, Ace O, Vera-Badillo F, Seruga B, Jordan J et al (2014) Prognostic relevance of receptor tyrosine kinase expression in breast cancer: a meta-analysis. Cancer Treat Rev 40(9):1048–1055

    Article  CAS  PubMed  Google Scholar 

  9. Kalykaki A, Agelaki S, Kallergi G, Xyrafas A, Mavroudis D, Georgoulias V (2014) Elimination of EGFR-expressing circulating tumor cells in patients with metastatic breast cancer treated with gefitinib. Cancer Chemother Pharmacol 73(4):685–693

    Article  CAS  PubMed  Google Scholar 

  10. Kuboki Y, Matsusaka S, Minowa S, Shibata H, Suenaga M, Shinozaki E et al (2013) Circulating tumor cell (CTC) count and epithelial growth factor receptor expression on CTCs as biomarkers for cetuximab efficacy in advanced colorectal cancer. Anticancer Res 33(9):3905–3910

    CAS  PubMed  Google Scholar 

  11. Balic M, Lin H, Williams A, Datar RH, Cote RJ (2012) Progress in circulating tumor cell capture and analysis: implications for cancer management. Expert Rev Mol Diagn 12(3):303–312

    Article  PubMed  PubMed Central  Google Scholar 

  12. Miller MC, Doyle GV, Terstappen LW (2010) Significance of circulating tumor cells detected by the CellSearch system in patients with metastatic breast colorectal and prostate cancer. J Oncol 2010:617421

    Article  PubMed  Google Scholar 

  13. Casavant BP, Mosher R, Warrick JW, Maccoux LJ, Berry SM, Becker JT et al (2013) A negative selection methodology using a microfluidic platform for the isolation and enumeration of circulating tumor cells. Methods 64(2):137–143

    Article  CAS  PubMed  Google Scholar 

  14. Farace F, Massard C, Vimond N, Drusch F, Jacques N, Billiot F et al (2011) A direct comparison of CellSearch and ISET for circulating tumour-cell detection in patients with metastatic carcinomas. Br J Cancer 105(6):847–853

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Zheng S, Lin HK, Lu B, Williams A, Datar R, Cote RJ et al (2011) 3D microfilter device for viable circulating tumor cell (CTC) enrichment from blood. Biomed Microdevices 13(1):203–213

    Article  PubMed  Google Scholar 

  16. van der Toom EE, Verdone JE, Gorin MA, Pienta KJ (2016) Technical challenges in the isolation and analysis of circulating tumor cells. Oncotarget 7:62754–62766

    Article  PubMed  PubMed Central  Google Scholar 

  17. Harouaka RA, Nisic M, Zheng SY (2013) Circulating tumor cell enrichment based on physical properties. J Lab Autom 18(6):455–468

    Article  CAS  PubMed  Google Scholar 

  18. Alix-Panabières C, Pantel K (2014) Challenges in circulating tumour cell research. Nat Rev Cancer 14(9):623–631

    Article  PubMed  Google Scholar 

  19. Schuur ER Rapid and simple isolation of circulating tumor cells for clinical and research applications using ScreenCell®. ScreenCell® devices: a flexible ctc platform

    Google Scholar 

  20. Desitter I, Guerrouahen BS, Benali-Furet N, Wechsler J, Janne PA, Kuang Y et al (2011) A new device for rapid isolation by size and characterization of rare circulating tumor cells. Anticancer Res 31(2):427–441

    PubMed  Google Scholar 

  21. Gaforio JJ, Serrano MJ, Sanchez-Rovira P, Sirvent A, Delgado-Rodriguez M, Campos M et al (2003) Detection of breast cancer cells in the peripheral blood is positively correlated with estrogen-receptor status and predicts for poor prognosis. Int J Cancer 107(6):984–990

    Article  CAS  PubMed  Google Scholar 

  22. Bayarri-Lara C, Ortega FG, Cueto Ladrón de Guevara A, Puche JL, Ruiz Zafra J, de Miguel-Pérez D et al (2016) Circulating tumor cells identify early recurrence in patients with non-small cell lung cancer undergoing radical resection. PLoS One 11(2):e0148659

    Article  PubMed  PubMed Central  Google Scholar 

  23. Karabacak NM, Spuhler PS, Fachin F, Lim EJ, Pai V, Ozkumur E et al (2014) Microfluidic, marker-free isolation of circulating tumor cells from blood samples. Nat Protoc 9(3):694–710

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Reinholz MM, Kitzmann KA, Tenner K, Hillman D, Dueck AC, Hobday TJ et al (2011) Cytokeratin-19 and mammaglobin gene expression in circulating tumor cells from metastatic breast cancer patients enrolled in North Central Cancer Treatment Group trials, N0234/336/436/437. Clin Cancer Res 17(22):7183–7193

    Article  CAS  PubMed  Google Scholar 

  25. Chen Y, Zou TN, Wu ZP, Zhou YC, Gu YL, Liu X et al (2011) Detection of cytokeratin 19, human mammaglobin, and carcinoembryonic antigen-positive circulating tumor cells by three-marker reverse transcription-PCR assay and its relation to clinical outcome in early breast cancer. Int J Biol Markers 25(2):59–68

    CAS  Google Scholar 

  26. Ning Y, Hanna DL, Zhang W, Mendez A, Yang D, El-Khoueiry R et al (2015) Cytokeratin-20 and survivin-expressing circulating tumor cells predict survival in metastatic colorectal cancer patients by a combined immunomagnetic qRT-PCR approach. Mol Cancer Ther 14(10):2401–2408

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Pal SK, He M, Wilson T, Liu X, Zhang K, Carmichael C et al (2015) Detection and phenotyping of circulating tumor cells in high-risk localized prostate cancer. Clin Genitourin Cancer 13(2):130–136

    Article  CAS  PubMed  Google Scholar 

  28. Armstrong AJ, Marengo MS, Oltean S, Kemeny G, Bitting RL, Turnbull JD et al (2011) Circulating tumor cells from patients with advanced prostate and breast cancer display both epithelial and mesenchymal markers. Mol Cancer Res 9(8):997–1007

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Satelli A, Batth IS, Brownlee Z, Rojas C, Meng QH, Kopetz S et al (2016) Potential role of nuclear PD-L1 expression in cell-surface vimentin positive circulating tumor cells as a prognostic marker in cancer patients. Sci Rep 6:28910

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Lindsay CR, Le Moulec S, Billiot F, Loriot Y, Ngo-Camus M, Vielh P et al (2016) Vimentin and Ki67 expression in circulating tumour cells derived from castrate-resistant prostate cancer. BMC Cancer 16:168

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Kallergi G, Papadaki MA, Politaki E, Mavroudis D, Georgoulias V, Agelaki S (2011) Epithelial to mesenchymal transition markers expressed in circulating tumour cells of early and metastatic breast cancer patients. Breast Cancer Res 13(3):R59

    Article  PubMed  PubMed Central  Google Scholar 

  32. Agelaki S, Kalykaki A, Markomanolaki H, Papadaki MA, Kallergi G, Hatzidaki D et al (2015) Efficacy of Lapatinib in therapy-resistant HER2-positive circulating tumor cells in metastatic breast cancer. PLoS One 10(6):e0123683

    Article  PubMed  PubMed Central  Google Scholar 

  33. Ady N, Morat L, Fizazi K, Soria JC, Mathieu MC, Prapotnich D et al (2004) Detection of HER-2/neu-positive circulating epithelial cells in prostate cancer patients. Br J Cancer 90(2):443–448

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Kalinsky K, Mayer JA, Xu X, Pham T, Wong KL, Villarin E et al (2015) Correlation of hormone receptor status between circulating tumor cells, primary tumor, and metastasis in breast cancer patients. Clin Transl Oncol 17(7):539–546

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Babayan A, Hannemann J, Spötter J, Müller V, Pantel K, Joosse SA (2013) Heterogeneity of estrogen receptor expression in circulating tumor cells from metastatic breast cancer patients. PLoS One 8(9):e75038

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Crespo M, van Dalum G, Ferraldeschi R, Zafeiriou Z, Sideris S, Lorente D et al (2015) Androgen receptor expression in circulating tumour cells from castration-resistant prostate cancer patients treated with novel endocrine agents. Br J Cancer 112(7):1166–1174

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Antonarakis ES, Lu C, Wang H, Luber B, Nakazawa M, Roeser JC et al (2014) AR-V7 and resistance to enzalutamide and abiraterone in prostate cancer. N Engl J Med 371(11):1028–1038

    Article  PubMed  PubMed Central  Google Scholar 

  38. Ning Y, Zhang W, Hanna DL, Yang D, Okazaki S, Berger MD et al (2016) Clinical relevance of EMT and stem-like gene expression in circulating tumor cells of metastatic colorectal cancer patients. Pharmacogenomics J. doi:10.1038/tpj.2016.62

  39. Hanssen A, Wagner J, Gorges TM, Taenzer A, Uzunoglu FG, Driemel C et al (2016) Characterization of different CTC subpopulations in non-small cell lung cancer. Sci Rep 6:28010

    Article  PubMed  PubMed Central  Google Scholar 

  40. Klinac D, Gray ES, Freeman JB, Reid A, Bowyer S, Millward M et al (2014) Monitoring changes in circulating tumour cells as a prognostic indicator of overall survival and treatment response in patients with metastatic melanoma. BMC Cancer 14:423

    Article  PubMed  PubMed Central  Google Scholar 

  41. Mostert B, Kraan J, Sieuwerts AM, van der Spoel P, Bolt-de V, Prager-van der Smissen WJ et al (2012) CD49f-based selection of circulating tumor cells (CTCs) improves detection across breast cancer subtypes. Cancer Lett 319(1):49–55

    Article  CAS  PubMed  Google Scholar 

  42. Nadal R, Ortega FG, Salido M, Lorente JA, Rodríguez-Rivera M, Delgado-Rodríguez M et al (2013) CD133 expression in circulating tumor cells from breast cancer patients: potential role in resistance to chemotherapy. Int J Cancer 133(10):2398–2407

    Article  CAS  PubMed  Google Scholar 

  43. Shi J, Li Y, Liang S, Zeng J, Liu G, Mu F et al (2016) Analysis of circulating tumor cells in colorectal liver metastasis patients before and after cryosurgery. Cancer Biol Ther 17(9):935–942

    Google Scholar 

  44. Kuhlmann JD, Wimberger P, Bankfalvi A, Keller T, Schöler S, Aktas B et al (2014) ERCC1-positive circulating tumor cells in the blood of ovarian cancer patients as a predictive biomarker for platinum resistance. Clin Chem 60(10):1282–1289

    Article  CAS  PubMed  Google Scholar 

  45. Dotan E, Alpaugh RK, Ruth K, Negin BP, Denlinger CS, Hall MJ et al (2016) Prognostic significance of MUC-1 in circulating tumor cells in patients with metastatic pancreatic adenocarcinoma. Pancreas 45(8):1131–1135

    Article  CAS  PubMed  Google Scholar 

  46. Nicolazzo C, Raimondi C, Mancini M, Caponnetto S, Gradilone A, Gandini O et al (2016) Monitoring PD-L1 positive circulating tumor cells in non-small cell lung cancer patients treated with the PD-1 inhibitor Nivolumab. Sci Rep 6:31726

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Mazel M, Jacot W, Pantel K, Bartkowiak K, Topart D, Cayrefourcq L et al (2015) Frequent expression of PD-L1 on circulating breast cancer cells. Mol Oncol 9(9):1773–1782

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Marchetti A, Del Grammastro M, Felicioni L, Malatesta S, Filice G, Centi I et al (2014) Assessment of EGFR mutations in circulating tumor cell preparations from NSCLC patients by next generation sequencing: toward a real-time liquid biopsy for treatment. PLoS One 9(8):e103883

    Article  PubMed  PubMed Central  Google Scholar 

  49. Mohamed Suhaimi NA, Foong YM, Lee DY, Phyo WM, Cima I, Lee EX et al (2015) Non-invasive sensitive detection of KRAS and BRAF mutation in circulating tumor cells of colorectal cancer patients. Mol Oncol 9(4):850–860

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Sakaizawa K, Goto Y, Kiniwa Y, Uchiyama A, Harada K, Shimada S et al (2012) Mutation analysis of BRAF and KIT in circulating melanoma cells at the single cell level. Br J Cancer 106(5):939–946

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Pestrin M, Salvianti F, Galardi F, De Luca F, Turner N, Malorni L et al (2015) Heterogeneity of PIK3CA mutational status at the single cell level in circulating tumor cells from metastatic breast cancer patients. Mol Oncol 9(4):749–757

    Article  CAS  PubMed  Google Scholar 

  52. Fernandez SV, Bingham C, Fittipaldi P, Austin L, Palazzo J, Palmer G et al (2014) TP53 mutations detected in circulating tumor cells present in the blood of metastatic triple negative breast cancer patients. Breast Cancer Res 16(5):445

    Article  PubMed  PubMed Central  Google Scholar 

  53. Danila DC, Anand A, Sung CC, Heller G, Leversha MA, Cao L et al (2011) TMPRSS2-ERG status in circulating tumor cells as a predictive biomarker of sensitivity in castration-resistant prostate cancer patients treated with abiraterone acetate. Eur Urol 60(5):897–904

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Punnoose EA, Ferraldeschi R, Szafer-Glusman E, Tucker EK, Mohan S, Flohr P et al (2015) PTEN loss in circulating tumour cells correlates with PTEN loss in fresh tumour tissue from castration-resistant prostate cancer patients. Br J Cancer 113(8):1225–1233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Onstenk W, Kraan J, Mostert B, Timmermans MM, Charehbili A, Smit VT et al (2015) Improved circulating tumor cell detection by a combined EpCAM and MCAM CellSearch enrichment approach in patients with breast cancer undergoing Neoadjuvant chemotherapy. Mol Cancer Ther 14(3):821–827

    Article  CAS  PubMed  Google Scholar 

  56. Gorges TM, Riethdorf S, von Ahsen O, Nastał YP, Röck K, Boede M et al (2016) Heterogeneous PSMA expression on circulating tumor cells - a potential basis for stratification and monitoring of PSMA-directed therapies in prostate cancer. Oncotarget 7:34930–34941

    Article  PubMed  PubMed Central  Google Scholar 

  57. Yokobori T, Iinuma H, Shimamura T, Imoto S, Sugimachi K, Ishii H 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(7):2059–2069

    Article  CAS  PubMed  Google Scholar 

  58. Ueo H, Sugimachi K, Gorges TM, Bartkowiak K, Yokobori T, Müller V et al (2015) Circulating tumour cell-derived plastin3 is a novel marker for predicting long-term prognosis in patients with breast cancer. Br J Cancer 112(9):1519–1526

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Abdallah EA, Fanelli MF, Souza E Silva V, Machado Netto MC, Gasparini Junior JL, Araújo DV et al (2016) MRP1 expression in CTCs confers resistance to irinotecan-based chemotherapy in metastatic colorectal cancer. Int J Cancer 139(4):890–898

    Article  CAS  PubMed  Google Scholar 

  60. Das M, Riess JW, Frankel P, Schwartz E, Bennis R, Hsieh HB et al (2012) ERCC1 expression in circulating tumor cells (CTCs) using a novel detection platform correlates with progression-free survival (PFS) in patients with metastatic non-small-cell lung cancer (NSCLC) receiving platinum chemotherapy. Lung Cancer 77(2):421–426

    Article  PubMed  Google Scholar 

  61. Liu S, Tian Z, Zhang L, Hou S, Hu S, Wu J et al (2016) Combined cell surface carbonic anhydrase 9 and CD147 antigens enable high-efficiency capture of circulating tumor cells in clear cell renal cell carcinoma patients. Oncotarget 7:59877–59891

    Article  PubMed  PubMed Central  Google Scholar 

  62. Ortega FG, Lorente JA, Garcia Puche JL, Ruiz MP, Sanchez-Martin RM, de Miguel-Pérez D et al (2015) miRNA in situ hybridization in circulating tumor cells--MishCTC. Sci Rep 5:9207

    Article  PubMed  PubMed Central  Google Scholar 

  63. Vona G, Sabile A, Louha M, Sitruk V, Romana S, Schütze K et al (2000) Isolation by size of epithelial tumor cells: a new method for the immunomorphological and molecular characterization of circulatingtumor cells. Am J Pathol 156(1):57–63

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Kim MS, Sim TS, Kim YJ, Kim SS, Jeong H, Park JM et al (2012) SSA-MOA: a novel CTC isolation platform using selective size amplification (SSA) and a multi-obstacle architecture (MOA) filter. Lab Chip 12(16):2874–2880

    Article  CAS  PubMed  Google Scholar 

  65. Hou HW, Warkiani ME, Khoo BL, Li ZR, Soo RA, Tan DS et al (2013) Isolation and retrieval of circulating tumor cells using centrifugal forces. Sci Rep 3:1259

    Article  PubMed  PubMed Central  Google Scholar 

  66. Weitz J, Kienle P, Lacroix J, Willeke F, Benner A, Lehnert T et al (1998) Dissemination of tumor cells in patients undergoing surgery for colorectal cancer. Clin Cancer Res 4(2):343–348

    CAS  PubMed  Google Scholar 

  67. Gascoyne PR, Noshari J, Anderson TJ, Becker FF (2009) Isolation of rare cells from cell mixtures by dielectrophoresis. Electrophoresis 30(8):1388–1398

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Müller V, Stahmann N, Riethdorf S, Rau T, Zabel T, Goetz A et al (2005) Circulating tumor cells in breast cancer: correlation to bone marrow micrometastases, heterogeneous response to systemic therapy and low proliferative activity. Clin Cancer Res 11(10):3678–3685

    Article  PubMed  Google Scholar 

  69. Rosenberg R, Gertler R, Friederichs J, Fuehrer K, Dahm M, Phelps R et al (2002) Comparison of two density gradient centrifugation systems for the enrichment of disseminated tumor cells in blood. Cytometry 49(4):150–158

    Article  CAS  PubMed  Google Scholar 

  70. Sollier E, Go DE, Che J, Gossett DR, O'Byrne S, Weaver WM et al (2014) Size-selective collection of circulating tumor cells using Vortex technology. Lab Chip 14(1):63–77

    Article  CAS  PubMed  Google Scholar 

  71. Hur SC, Henderson-MacLennan NK, McCabe ER, Di Carlo D (2011) Deformability-based cell classification and enrichment using inertial microfluidics. Lab Chip 11(5):912–920

    Article  CAS  PubMed  Google Scholar 

  72. Fischer JC, Niederacher D, Topp SA, Honisch E, Schumacher S, Schmitz N et al (2013) Diagnostic leukapheresis enables reliable detection of circulating tumor cells of nonmetastatic cancer patients. Proc Natl Acad Sci U S A 110(41):16580–16585

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Eifler RL, Lind J, Falkenhagen D, Weber V, Fischer MB, Zeillinger R (2011) Enrichment of circulating tumor cells from a large blood volume using leukapheresis and elutriation: proof of concept. Cytometry B Clin Cytom 80(2):100–111

    Article  PubMed  Google Scholar 

  74. Hayes DF, Cristofanilli M, Budd GT, Ellis MJ, Stopeck A, Miller MC et al (2006) 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 12(14):4218–4224

    Article  CAS  PubMed  Google Scholar 

  75. Cohen SJ, Punt CJ, Iannotti N, Saidman BH, Sabbath KD, Gabrail NY et al (2008) Relationship of circulating tumor cells to tumor response, progression-free survival, and overall survival in patients with metastatic colorectal cancer. J Clin Oncol 26(19):3213–3221

    Article  PubMed  Google Scholar 

  76. de Bono JS, Scher HI, Montgomery RB, Parker C, Miller MC, Tissing H et al (2008) Circulating tumor cells predict survival benefit from treatment in metastatic castration-resistant prostate cancer. Clin Cancer Res 14(19):6302–6309

    Article  PubMed  Google Scholar 

  77. Riethdorf S, Fritsche H, Müller V, Rau T, Schindlbeck C, Rack B et al (2007) Detection of circulating tumor cells in peripheral blood of patients with metastatic breast cancer: a validation study of the CellSearch system. Clin Cancer Res 13(3):920–928

    Article  CAS  PubMed  Google Scholar 

  78. Todenhöfer T, Hennenlotter J, Feyerabend S, Aufderklamm S, Mischinger J, Kühs U et al (2012) Preliminary experience on the use of the Adnatest® system for detection of circulating tumor cells in prostate cancer patients. Anticancer Res 32(8):3507–3513

    PubMed  Google Scholar 

  79. Müller V, Riethdorf S, Rack B, Janni W, Fasching PA, Solomayer E et al (2012) Prognostic impact of circulating tumor cells assessed with the CellSearch System™ and AdnaTest Breast™ in metastatic breast cancer patients: the DETECT study. Breast Cancer Res 14(4):R118

    Article  PubMed  PubMed Central  Google Scholar 

  80. Talasaz AH, Powell AA, Huber DE, Berbee JG, Roh KH, Yu W et al (2009) Isolating highly enriched populations of circulating epithelial cells and other rare cells from blood using a magnetic sweeper device. Proc Natl Acad Sci U S A 106(10):3970–3975

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  81. Sequist LV, Nagrath S, Toner M, Haber DA, Lynch TJ (2009) The CTC-chip: an exciting new tool to detect circulating tumor cells in lung cancer patients. J Thorac Oncol 4(3):281–283

    Article  PubMed  PubMed Central  Google Scholar 

  82. Stott SL, Hsu CH, Tsukrov DI, Yu M, Miyamoto DT, Waltman BA et al (2010) Isolation of circulating tumor cells using a microvortex-generating herringbone-chip. Proc Natl Acad Sci U S A 107(43):18392–18397

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  83. Nagrath S, Sequist LV, Maheswaran S, Bell DW, Irimia D, Ulkus L et al (2007) Isolation of rare circulating tumour cells in cancer patients by microchip technology. Nature 450(7173):1235–1239

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  84. Saliba AE, Saias L, Psychari E, Minc N, Simon D, Bidard FC et al (2010) Microfluidic sorting and multimodal typing of cancer cells in self-assembled magnetic arrays. Proc Natl Acad Sci U S A 107(33):14524–14529

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  85. Harb W, Fan A, Tran T, Danila DC, Keys D, Schwartz M et al (2013) Mutational analysis of circulating tumor cells using a novel microfluidic collection device and qPCR assay. Transl Oncol 6(5):528–538

    Article  PubMed  PubMed Central  Google Scholar 

  86. Chen JF, Zhu Y, Lu YT, Hodara E, Hou S, Agopian VG et al (2016) Clinical applications of NanoVelcro rare-cell assays for detection and characterization of circulating tumor cells. Theranostics 6(9):1425–1439

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  87. Kirby BJ, Jodari M, Loftus MS, Gakhar G, Pratt ED, Chanel-Vos C et al (2012) Functional characterization of circulating tumor cells with a prostate-cancer-specific microfluidic device. PLoS One 7(4):e35976

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  88. Fabbri F, Carloni S, Zoli W, Ulivi P, Gallerani G, Fici P 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):225–231

    Article  CAS  PubMed  Google Scholar 

  89. Saucedo-Zeni N, Mewes S, Niestroj R, Gasiorowski L, Murawa D, Nowaczyk P 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):1241–1250

    PubMed  PubMed Central  Google Scholar 

  90. Wu Y, Deighan CJ, Miller BL, Balasubramanian P, Lustberg MB, Zborowski M et al (2013) Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodology. Methods 64(2):169–182

    Article  CAS  PubMed  Google Scholar 

  91. Yang L, Lang JC, Balasubramanian P, Jatana KR, Schuller D, Agrawal A 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):521–534

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  92. Ozkumur E, Shah AM, Ciciliano JC, Emmink BL, Miyamoto DT, Brachtel E et al (2013) Inertial focusing for tumor antigen-dependent and -independent sorting of rare circulating tumor cells. Sci Transl Med 5(179):179ra147

    Article  Google Scholar 

  93. Naume B, Borgen E, Tøssvik S, Pavlak N, Oates D, Nesland JM (2004) Detection of isolated tumor cells in peripheral blood and in BM: evaluation of a new enrichment method. Cytotherapy 6(3):244–252

    Article  CAS  PubMed  Google Scholar 

  94. Mostert B, Sleijfer S, Foekens JA, Gratama JW (2009) Circulating tumor cells (CTCs): detection methods and their clinical relevance in breast cancer. Cancer Treat Rev 35(5):463–474

    Article  CAS  PubMed  Google Scholar 

  95. Zuccolo J, Unruh TL, Deans JP (2009) Efficient isolation of highly purified tonsil B lymphocytes using RosetteSep with allogeneic human red blood cells. BMC Immunol 10:30

    Article  PubMed  PubMed Central  Google Scholar 

  96. Werner SL, Graf RP, Landers M, Valenta DT, Schroeder M, Greene SB et al (2015) Analytical validation and capabilities of the Epic CTC platform:enrichment-freecirculating tumour cell detection and characterization. J Circulating Biomarkers 4, 3

    Google Scholar 

  97. Krivacic RT, Ladanyi A, Curry DN, Hsieh HB, Kuhn P, Bergsrud DE et al (2004) A rare-cell detector for cancer. Proc Natl Acad Sci U S A 101(29):10501–10504

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  98. Campton DE, Ramirez AB, Nordberg JJ, Drovetto N, Clein AC, Varshavskaya P et al (2015) High-recovery visual identification and single-cell retrieval of circulating tumor cells for genomic analysis using a dual-technology platform integrated with automated immunofluorescence staining. BMC Cancer 15:360

    Article  PubMed  PubMed Central  Google Scholar 

  99. Alix-Panabières C, Pantel K (2015) Liquid biopsy in cancer patients: advances in capturing viable CTCs for functional studies using the EPISPOT assay. Expert Rev Mol Diagn 15(11):1411–1417

    Article  PubMed  Google Scholar 

  100. Tulley S, Zhao Q, Dong H, Pearl ML, Chen WT (2016) Vita-Assay™ method of enrichment and identification of circulating cancer cells/circulating tumor cells (CTCs). Methods Mol Biol 1406:107–119

    Article  CAS  PubMed  Google Scholar 

  101. Baccelli I, Schneeweiss A, Riethdorf S, Stenzinger A, Schillert A, Vogel V et al (2013) Identification of a population of blood circulating tumor cells from breast cancer patients that initiates metastasis in a xenograft assay. Nat Biotechnol 31(6):539–544

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. GENYO. Centre for Genomics and Oncological Research, Pfizer, University of Granada, Andalusian Regional Government, Av. de la Ilustración 114, 18016, Granada, Spain

    M. J. Alvarez Cubero, J. A. Lorente, I. Robles-Fernandez, A. Rodriguez-Martinez & M. J. Serrano

  2. Laboratory of Genetic Identification, University of Granada-Dept. of Legal Medicine - Faculty of Medicine, Granada, 18016, Spain

    J. A. Lorente & A. Rodriguez-Martinez

  3. Integral Oncology Division, Clinical University Hospitals of Granada, Av. de las Fuerzas Armadas, 2, 18014, Granada, Spain

    J. L. Puche & M. J. Serrano

Authors
  1. M. J. Alvarez Cubero
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. A. Lorente
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. I. Robles-Fernandez
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. Rodriguez-Martinez
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. L. Puche
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M. J. Serrano
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. J. Alvarez Cubero .

Editor information

Editors and Affiliations

  1. Division of Hematology Oncology, Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, San Francisco, California, USA

    Mark Jesus M. Magbanua

  2. Division of Hematology Oncology, Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, San Francisco, California, USA

    John W. Park

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Alvarez Cubero, M.J., Lorente, J.A., Robles-Fernandez, I., Rodriguez-Martinez, A., Puche, J.L., Serrano, M.J. (2017). Circulating Tumor Cells: Markers and Methodologies for Enrichment and Detection. In: M. Magbanua, M., W. Park, J. (eds) Circulating Tumor Cells. Methods in Molecular Biology, vol 1634. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7144-2_24

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7144-2_24

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7143-5

  • Online ISBN: 978-1-4939-7144-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation