Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

12 Issues per year


IMPACT FACTOR 2017: 3.556

CiteScore 2017: 2.34

SCImago Journal Rank (SJR) 2017: 1.114
Source Normalized Impact per Paper (SNIP) 2017: 1.188

Online
ISSN
1437-4331
See all formats and pricing
More options …
Volume 52, Issue 2

Issues

Enrichment and enumeration of circulating tumor cells by efficient depletion of leukocyte fractions

Shiyang Wu
  • SurExam Bio-Tech Co. Ltd., Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Zhiming Liu
  • SurExam Bio-Tech Co. Ltd., Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Suyan Liu
  • SurExam Bio-Tech Co. Ltd., Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Li Lin
  • SurExam Bio-Tech Co. Ltd., Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Weiwei Yang
  • SurExam Bio-Tech Co. Ltd., Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Jiasen Xu
  • Corresponding author
  • SurExam Bio-Tech Co. Ltd., Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2013-09-09 | DOI: https://doi.org/10.1515/cclm-2013-0558

An erratum for this article can be found here: https://doi.org/10.1515/cclm-2015-9999

Abstract

Background: Enumeration and characterization of circulating tumor cells (CTCs) can provide information on patient prognosis and treatment efficacy. However, CTCs are rare, making their isolation a major technological challenge. We developed a technique for enrichment, and subsequent characterization of CTCs based on efficient depletion of human leukocytes.

Methods: The technique (CanPatrolTM CTC enrichment) we developed is based on red blood cell lysis to remove erythrocytes, followed by depletion of CD45+ leukocytes using a magnetic bead separation method, and subsequent isolation of CTCs by virtue of their larger size, compared with leukocytes. We also demonstrated that fluorescence in situ hybridization (FISH) and genetic abnormalities analysis could be performed on the isolated CTCs.

Results: The spiking experiments showed that the average efficacy of leukocytes depletion was 99.98% and the average tumor cells recovery was not lower than 80%. FISH could be used to perform ALK gene rearrangement analysis on the collected NCI-H2228 cells, and EGFR Exon 19 deletion was detected by PCR-based analysis in isolated HCC827 cells. The in vivo feasibility of this technique had been demonstrated in patients with non-small cell lung cancer, breast, colon, and esophageal cancers. CTCs were detected in 13 of 59 blood samples. Tumor microemboli was also detected in three breast cancer samples.

Conclusions: The technique we developed allowed isolation and characterization of circulating epithelial tumor cells that do not express classical epithelial antigens. This potentially leads to a more accurate enumeration of the number of CTCs and is suitable for application to a broad range of cancers.

Keywords: circulating tumor cells (CTCs); depletion of CD45+ leukocytes; fluorescence in situ hybridization (FISH); genetic abnormalities analysis; isolation of CTCs by size

References

  • 1.

    Gupta GP, Massagué J. Cancer metastasis: building a framework. Cell 2006;127:679–95.CrossrefPubMedGoogle Scholar

  • 2.

    Schuster R, Bechrakis NE, Stroux A, Busse A, Schmittel A, Scheibenbogen C, et al. Circulating tumor cells as prognostic factor for distant metastases and survival in patients with primary uveal melanoma. Clin Cancer Res 2007;13:1171–8.PubMedCrossrefGoogle Scholar

  • 3.

    Taback B, Chan AD, Kuo CT, Bostick PJ, Wang HJ, Giuliano AE, et al. Detection of occult metastatic breast cancer cells in blood by a multimolecular marker assay: correlation with clinical stage of disease. Cancer Res 2001;61:8845–50.PubMedGoogle Scholar

  • 4.

    Liu Z, Jiang M, Zhao J, Ju H. Circulating tumor cells in perioperative esophageal cancer patients: quantitative assay system and potential clinical utility. Clin Cancer Res 2007;13:2992–7.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 5.

    Cohen SJ, Alpaugh RK, Gross S, O’Hara SM, Smirnov DA, Terstappen LW, et al. Isolation and characterization of circulating tumor cells in patients with metastatic colorectal cancer. Clin Colorectal Cancer 2006;6:125–32.CrossrefPubMedGoogle Scholar

  • 6.

    Cristofanilli M, Hayes DF, Budd GT, Ellis MJ, Stopeck A, Reuben JM, et al. Circulating tumor cells: a novel prognostic factor for newly diagnosed metastatic breast cancer. J Clin Oncol 2005;23:1420–30.CrossrefPubMedGoogle Scholar

  • 7.

    Cohen SJ, Punt CJ, Iannotti N, Saidman BH, Sabbath KD, Gabrail NY, et al. Relationship of circulating tumor cells to tumor response, progression-free survival, and overall survival in patients with metastatic colorectal cancer. J Clin Oncol 2008;26:3213–21.PubMedCrossrefGoogle Scholar

  • 8.

    de Bono JS, Scher HI, Montgomery RB, Parker C, Miller MC, Tissing H, et al. Circulating tumor cells predict survival benefit from treatment in metastatic castration-resistant prostate cancer. Clin Cancer Res 2008;14:6302–9.CrossrefGoogle Scholar

  • 9.

    Krebs MG, Sloane R, Priest L, Lancashire L, Hou JM, Greystoke A, et al. Evaluation and prognostic significance of circulating tumor cells in patients with non-small-cell lung cancer. J Clin Oncol 2011;29:1556–63.PubMedCrossrefGoogle Scholar

  • 10.

    Yu M, Stott S, Toner M, Maheswaran S, Haber D. Circulating tumor cells: approaches to isolation and characterization. J Cell Biol 2011;192:373–82.Web of ScienceGoogle Scholar

  • 11.

    Riethdorf S, Fritsche H, Müller V, Rau T, Schindlbeck C, Rack B, et al. Detection of circulating tumor cells in peripheral blood of patients with metastatic breast cancer: a validation study of the CellSearch system. Clin Cancer Res 2007;13:920–8.PubMedCrossrefGoogle Scholar

  • 12.

    Paterlini-Brechot P, Benali NL. Circulating tumor cells (CTC) detection: clinical impact and future directions. Cancer Letters 2007;253:180–204.Web of ScienceGoogle Scholar

  • 13.

    Li GQ, Luo XD, He JY, Zhu ZY, Yu G, Qin HJ, et al. A novel liquidchip platform for simultaneous detection of 70 alleles of DNA somatic mutations on EGFR, KRAS, BRAF and PIK3CA from formalin-fixed and paraffin-embedded slides containing tumor tissue. Clin Chem Lab Med 2011;49:191–5.CrossrefWeb of ScienceGoogle Scholar

  • 14.

    Maheswaran S, Sequist LV, Nagrath S, Ulkus L, Brannigan B, Collura CV, et al. Detection of mutations in EGFR in circulating lung-cancer cells. N Engl J Med 2008;359:366–77.Web of ScienceGoogle Scholar

  • 15.

    Klein CA, Schmidt-Kittler O, Schardt JA, Pantel K, Speicher MR, Riethmüller G. Comparative genomic hybridization, loss of heterozygosity, and DNA sequence analysis of single cells. Proc Natl Acad Sci USA 1999;96:4494–9.CrossrefGoogle Scholar

  • 16.

    Kraus J, Pantel K, Pinkel D, Albertson DG, Speicher MR. High-resolution genomic profiling of occult micrometastatic tumor cells. Genes Chromosomes Cancer 2003;36:159–66.CrossrefPubMedGoogle Scholar

  • 17.

    Liu ZA, Fusi A, Klopocki E, Schmitte A, Tinhofer I, Nonnenmacher A, et al. Negative enrichment by immunomagnetic nanobeads for unbiased characterization of circulating tumor cells from peripheral blood of cancer patients. J Transl Med 2011;9:70–8.Web of SciencePubMedCrossrefGoogle Scholar

  • 18.

    Parkinson DR, Dracopoli N, Petty BG, Compton C, Cristofanilli M, Deisseroth A, et al. Considerations in the development of circulating tumor cell technology for clinical use. J Transl Med 2012;10:138–58.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 19.

    Stolpe A, Pantel K, Sleijfer S, Terstappen LW, Toonder J. Circulating tumor cell isolation and diagnostics: toward routine clinical use. Cancer Res 2011;71:5955–60.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 20.

    Sieuwerts AM, Kraan J, Bolt J, van der Spoel P, Elstrodt F, Schutte M, et al. Anti-epithelial cell adhesion molecule antibodies and the detection of circulating normal-like breast tumor cells. J Natl Cancer Inst 2009;101:61–6.CrossrefGoogle Scholar

  • 21.

    Vona G, Sabile A, Louha M, Sitruk V, Romana S, Schutze K, et al. Isolation by size of epithelial tumor cells. Am J Pathol 2000;156:57–63.Google Scholar

About the article

Corresponding author: Jiasen Xu, SurExam Bio-Tech Co. Ltd., Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China, E-mail:


Received: 2013-05-27

Accepted: 2013-08-03

Published Online: 2013-09-09

Published in Print: 2014-02-01


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 52, Issue 2, Pages 243–251, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2013-0558.

Export Citation

©2014 by Walter de Gruyter Berlin Boston.Get Permission

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

[1]
[2]
Liang-Chun Yin, Zhen-Chao Luo, Yan-Xin Gao, Yang Li, Qing Peng, and Yi Gao
BioMed Research International, 2018, Volume 2018, Page 1
[4]
MAOWEN GUO, XIAOTIAN LI, SHAOHUA ZHANG, HUA SONG, WENHUI ZHANG, XUEYI SHANG, YULING ZHENG, HUA JIANG, QINGYU LV, YONGQIANG JIANG, and HUAIJIE HAO
International Journal of Oncology, 2015, Volume 46, Number 1, Page 281
[5]
Wei Zhang, Wenjie Xia, Zhengye Lv, Chao Ni, Yin Xin, and Liu Yang
Cellular Physiology and Biochemistry, 2017, Volume 41, Number 2, Page 755
[6]
Chuanli Ren, Chongxu Han, Deyuan Fu, Daxin Wang, Hui Chen, Yong Chen, and Ming Shen
International Journal of Cancer, 2016, Volume 138, Number 7, Page 1586

Comments (0)

Please log in or register to comment.
Log in