Detection of Circulating Tumor Cells in Blood of Metastatic Breast Cancer Patients Using a Combination of Cytokeratin and EpCAM Antibodies

Overview

Journal BMC Cancer

Publisher Biomed Central

Specialty Oncology

Date 2012 Jun 1

PMID 22646670

Citations 30

Authors

Ulrike Weissenstein

Agnes Schumann

Marcus Reif

Susanne Link

Ulrike D Toffol-Schmidt

Peter Heusser

Affiliations

Soon will be listed here.

Abstract

Background: Circulating tumor cells (CTCs) are detectable in peripheral blood of metastatic breast cancer patients (MBC). In this paper we evaluate a new CTC separation method based on a combination of anti-EpCAM- and anti-cytokeratin magnetic cell separation with the aim to improve CTC detection with low target antigen densities.

Methods: Blood samples of healthy donors spiked with breast cancer cell line HCC1937 were used to determine accuracy and precision of the method. 10 healthy subjects were examined to evaluate specificity. CTC counts in 59 patients with MBC were measured to evaluate the prognostic value on overall survival.

Results: Regression analysis of numbers of recovered vs. spiked HCC1937 cells yielded a coefficient of determination of R(2) = 0.957. The average percentage of cell recovery was 84%. The average within-run coefficient of variation for spiking of 185, 85 and 30 cells was 14%. For spiking of 10 cells the within-run CV was 30%. No CTCs were detected in blood of 10 healthy subjects examined. A standard threshold of 5 CTC/7.5 ml blood as a cut-off point between risk groups led to a highly significant prognostic marker (p < 0.001). To assess the prognostic value of medium CTC levels we additionally considered a low (CTC-L: 0 CTC), a medium (CTC-M: 1-4 CTC) and a high risk group (CTC-H: ≥5 CTC). The effect of this CTC-LMH marker on overall survival was significant as well (p < 0.001). A log-ratio test performed to compare the model with 3 vs. the model with 2 risk groups rejected the model with 2 risk groups (p = 0.026). For CTC as a count variable, we propose an offset reciprocal transformation 1/(1 + x) for overall survival prediction (p < 0.001).

Conclusions: We show that our CTC detection method is feasible and leads to accurate and reliable results. Our data suggest that a refined differentiation between patients with different CTC levels is reasonable.

Citing Articles

Detection, significance and potential utility of circulating tumor cells in clinical practice in breast cancer (Review).

Rusnakova D, Aziri R, Dubovan P, Jurik M, Mego M, Pindak D Oncol Lett. 2024; 29(1):10.

PMID: 39492933 PMC: 11526295. DOI: 10.3892/ol.2024.14756.

Factors influencing the diagnostic and prognostic values of circulating tumor cells in breast cancer: a meta-analysis of 8,935 patients.

Zhao H, Wang L, Fang C, Li C, Zhang L Front Oncol. 2023; 13:1272788.

PMID: 38090481 PMC: 10711619. DOI: 10.3389/fonc.2023.1272788.

Circulating tumor biomarkers in early-stage breast cancer: characteristics, detection, and clinical developments.

Qiu J, Qian D, Jiang Y, Meng L, Huang L Front Oncol. 2023; 13:1288077.

PMID: 37941557 PMC: 10628786. DOI: 10.3389/fonc.2023.1288077.

Updated Clinical Perspectives and Challenges of Chimeric Antigen Receptor-T Cell Therapy in Colorectal Cancer and Invasive Breast Cancer.

Cao Y, Efetov S, He M, Fu Y, Beeraka N, Zhang J Arch Immunol Ther Exp (Warsz). 2023; 71(1):19.

PMID: 37566162 DOI: 10.1007/s00005-023-00684-x.

Silibinin exhibits anti-tumor effects in a breast cancer stem cell model by targeting stemness and induction of differentiation and apoptosis.

Firouzi J, Sotoodehnejadnematalahi F, Shokouhifar A, Rahimi M, Sodeifi N, Sahranavardfar P Bioimpacts. 2022; 12(5):415-429.

PMID: 36381630 PMC: 9596878. DOI: 10.34172/bi.2022.23336.

References

Muller V, Stahmann N, Riethdorf S, Rau T, Zabel T, Goetz A . Circulating tumor cells in breast cancer: correlation to bone marrow micrometastases, heterogeneous response to systemic therapy and low proliferative activity. Clin Cancer Res. 2005; 11(10):3678-85. DOI: 10.1158/1078-0432.CCR-04-2469. View

Riethdorf S, Fritsche H, Muller V, Rau T, Schindlbeck C, Rack B . 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(3):920-8. DOI: 10.1158/1078-0432.CCR-06-1695. View

Borgen E, Naume B, Nesland J, Kvalheim G, Beiske K, Fodstad O . Standardization of the immunocytochemical detection of cancer cells in BM and blood: I. establishment of objective criteria for the evaluation of immunostained cells. Cytotherapy. 2010; 1(5):377-88. DOI: 10.1080/0032472031000141283. View

Campos M, Prior C, Warleta F, Zudaire I, Ruiz-Mora J, Catena R . Phenotypic and genetic characterization of circulating tumor cells by combining immunomagnetic selection and FICTION techniques. J Histochem Cytochem. 2008; 56(7):667-75. PMC: 2430160. DOI: 10.1369/jhc.2008.951111. View

Munz M, Baeuerle P, Gires O . The emerging role of EpCAM in cancer and stem cell signaling. Cancer Res. 2009; 69(14):5627-9. DOI: 10.1158/0008-5472.CAN-09-0654. View

Hayes D, Ethier S, Lippman M . New guidelines for reporting of tumor marker studies in breast cancer research and treatment: REMARK. Breast Cancer Res Treat. 2006; 100(2):237-8. DOI: 10.1007/s10549-006-9253-5. View

Nole F, Munzone E, Zorzino L, Minchella I, Salvatici M, Botteri E . Variation of circulating tumor cell levels during treatment of metastatic breast cancer: prognostic and therapeutic implications. Ann Oncol. 2007; 19(5):891-7. DOI: 10.1093/annonc/mdm558. View

Ross A, Cooper B, Lazarus H, Mackay W, Moss T, Ciobanu N . Detection and viability of tumor cells in peripheral blood stem cell collections from breast cancer patients using immunocytochemical and clonogenic assay techniques. Blood. 1993; 82(9):2605-10. View

Cristofanilli M, Budd G, Ellis M, Stopeck A, Matera J, Miller M . Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med. 2004; 351(8):781-91. DOI: 10.1056/NEJMoa040766. View

10.

Tibbe A, Miller M, Terstappen L . Statistical considerations for enumeration of circulating tumor cells. Cytometry A. 2007; 71(3):154-62. DOI: 10.1002/cyto.a.20369. View

11.

Pierce B, Ballard-Barbash R, Bernstein L, Baumgartner R, Neuhouser M, Wener M . Elevated biomarkers of inflammation are associated with reduced survival among breast cancer patients. J Clin Oncol. 2009; 27(21):3437-44. PMC: 2717751. DOI: 10.1200/JCO.2008.18.9068. View

12.

De Giorgi U, Valero V, Rohren E, Mego M, Doyle G, Miller M . Circulating tumor cells and bone metastases as detected by FDG-PET/CT in patients with metastatic breast cancer. Ann Oncol. 2009; 21(1):33-9. DOI: 10.1093/annonc/mdp262. View

13.

Gaforio J, Serrano M, Sanchez-Rovira P, Sirvent A, Delgado-Rodriguez M, Campos M . Detection of breast cancer cells in the peripheral blood is positively correlated with estrogen-receptor status and predicts for poor prognosis. Int J Cancer. 2003; 107(6):984-90. DOI: 10.1002/ijc.11479. View

14.

McShane L, Altman D, Sauerbrei W, Taube S, Gion M, Clark G . Reporting recommendations for tumor marker prognostic studies (remark). Exp Oncol. 2006; 28(2):99-105. View

15.

Songun I, Litvinov S, van de Velde C, Pals S, Hermans J, van Krieken J . Loss of Ep-CAM (CO17-1A) expression predicts survival in patients with gastric cancer. Br J Cancer. 2005; 92(9):1767-72. PMC: 2362035. DOI: 10.1038/sj.bjc.6602519. View

16.

Allard W, Matera J, Miller M, Repollet M, Connelly M, Rao C . Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin Cancer Res. 2004; 10(20):6897-904. DOI: 10.1158/1078-0432.CCR-04-0378. View

17.

Helo P, Cronin A, Danila D, Wenske S, Gonzalez-Espinoza R, Anand A . Circulating prostate tumor cells detected by reverse transcription-PCR in men with localized or castration-refractory prostate cancer: concordance with CellSearch assay and association with bone metastases and with survival. Clin Chem. 2009; 55(4):765-73. PMC: 2782624. DOI: 10.1373/clinchem.2008.117952. View

18.

Wirtschafter A, Benninger M, Moss T, Umiel T, Blazoff K, Worsham M . Micrometastatic tumor detection in patients with head and neck cancer: a preliminary report. Arch Otolaryngol Head Neck Surg. 2002; 128(1):40-3. DOI: 10.1001/archotol.128.1.40. View

19.

Rao C, Chianese D, Doyle G, Miller M, Russell T, Sanders Jr R . Expression of epithelial cell adhesion molecule in carcinoma cells present in blood and primary and metastatic tumors. Int J Oncol. 2005; 27(1):49-57. View

20.

Liu M, Shields P, Warren R, Cohen P, Wilkinson M, Ottaviano Y . Circulating tumor cells: a useful predictor of treatment efficacy in metastatic breast cancer. J Clin Oncol. 2009; 27(31):5153-9. PMC: 4879719. DOI: 10.1200/JCO.2008.20.6664. View