Advertisement

Flow Cytometric Detection of Circulating Tumor Cells Using a Candidate Stem Cell Marker, p75 Neurotrophin Receptor (p75NTR)

  • Tomoyuki OkumuraEmail author
  • Tetsuji Yamaguchi
  • Toru Watanabe
  • Takuya Nagata
  • Yutaka Shimada
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1634)

Abstract

The most widely studied detection for circulating tumor cells (CTCs) in peripheral blood of cancer patients has been based on immunomagnetic enrichment using antibodies against epithelial cell adhesion molecule (EpCAM), which is overexpressed in epithelial cells. A neurotrophin receptor p75 (p75NTR) is expressed in a candidate stem cell fraction in esophageal squamous cell carcinoma (ESCC), which shows significantly higher colony formation, enhanced tumor formation in mice, along with strong expression of epithelial mesenchymal transition-related genes. Here, we describe a method to detect CTCs in ESCC based on the combined expression of EpCAM and p75NTR using flow cytometry, demonstrating the feasibility of expression analysis of multiple cell surface markers in viable cells.

Key words

Esophageal cancer Circulating tumor cells Cancer stem cells Flow cytometry p75NTR 

Notes

Acknowledgment

This work was supported by JSPS KAKENHI Grant Number JP15K10088.

References

  1. 1.
    Rice TW, Rusch VW, Apperson-Hansen C, Allen MS, Chen L-Q, Hunter JG et al (2009) Worldwide esophageal cancer collaboration. Dis Esophagus 22:1–8CrossRefPubMedGoogle Scholar
  2. 2.
    Thallinger CMR, Raderer M, Hejna M (2011) Esophageal cancer: a critical evaluation of systemic second-line therapy. J Clin Oncol 29:4709–4714CrossRefPubMedGoogle Scholar
  3. 3.
    Hughes AD, King MR (2012) Nanobiotechnology for the capture and manipulation of circulating tumor cells. Wiley Interdiscip Rev Nanomed Nanobiotechnol 4:291–309CrossRefPubMedGoogle Scholar
  4. 4.
    Ross AH, Herlyn D, Iliopoulos D, Koprowski H (1986) Isolation and characterization of a carcinoma-associated antigen. Biochem Biophys Res Commun 135:297–303CrossRefPubMedGoogle Scholar
  5. 5.
    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 35:463–474CrossRefPubMedGoogle Scholar
  6. 6.
    Hayes DF, Smerage J (2008) Is there a role for circulating tumor cells in the management of breast cancer? Clin Cancer Res 14:3646–3650CrossRefPubMedGoogle Scholar
  7. 7.
    Reya T, Morrison SJ, Clarke MF, Weissman IL (2001) Stem cells, cancer, and cancer stem cells. Nature 414:105–111CrossRefPubMedGoogle Scholar
  8. 8.
    Clarke MF, Dick JE, Dirks PB, Eaves CJ, Jamieson CH, Jones DL et al (2006) Cancer stem cells-perspectives on current status and future directions: AACR Workshop on cancer stem cells. Cancer Res 66:9339–9344CrossRefPubMedGoogle Scholar
  9. 9.
    Grover PK, Cummins AG, Price TJ, Roberts-Thomson IC, Hardingham JE (2014) Circulating tumour cells: the evolving concept and the inadequacy of their enrichment by EpCAM-based methodology for basic and clinical cancer research. Ann Oncol 25:1506–1516CrossRefPubMedGoogle Scholar
  10. 10.
    Dean M, Fojo T, Bates S (2005) Tumour stem cells and drug resistance. Nat Rev Cancer 5:275–284CrossRefPubMedGoogle Scholar
  11. 11.
    Schatton T, Frank MH (2008) Cancer stem cells and human malignant melanoma. Pigment Cell Melanoma Res 21:39–55CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Wicha MS, Hayes DF (2011) Circulating tumor cells: not all detected cells are bad and not all bad cells are detected. J Clin Oncol 29:1508–1511CrossRefPubMedGoogle Scholar
  13. 13.
    Okumura T, Tsunoda S, Mori Y, Ito T, Kikuchi K, Wang TC et al (2006) The biological role of the low-affinity p75 neurotrophin receptor in esophageal squamous cell carcinoma. Clin Cancer Res 12:5096–5103CrossRefPubMedGoogle Scholar
  14. 14.
    Huang S-D, Yuan Y, Liu X-H, Gong D-J, Bai C-G, Wang F et al (2009) Self-renewal and chemotherapy resistance of p75NTR positive cells in esophageal squamous cell carcinomas. BMC Cancer 9:9CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Yamaguchi T, Okumura T, Hirano K, Watanabe T, Nagata T, Shimada Y et al (2016) p75 neurotrophin receptor expression is a characteristic of the mitotically quiescent cancer stem cell population present in esophageal squamous cell carcinoma. Int J Oncol 48:1943–1954PubMedGoogle Scholar
  16. 16.
    Yamaguchi T, Okumura T, Hirano K, Watanabe T, Nagata T, Shimada Y et al (2016) Detection of circulating tumor cells by p75NTR expression in patients with esophageal cancer. World J Surg Oncol 14:40CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Tomoyuki Okumura
    • 1
    Email author
  • Tetsuji Yamaguchi
    • 1
  • Toru Watanabe
    • 1
  • Takuya Nagata
    • 1
  • Yutaka Shimada
    • 2
  1. 1.Department of Surgery and Science, Graduate School of Medicine and Pharmaceutical Sciences for ResearchUniversity of ToyamaToyama CityJapan
  2. 2.Department of Nanobio Drug Discovery, Graduate School of Pharmaceutical SciencesKyoto UniversityKyotoJapan

Personalised recommendations