Detection of EpCAM-Negative and Cytokeratin-Negative Circulating Tumor Cells in Peripheral Blood

Overview

Journal J Oncol

Specialty Oncology

Date 2011 May 18

PMID 21577258

Citations 116

Authors

Stephen D Mikolajczyk

Lisa S Millar

Pavel Tsinberg

Stephen M Coutts

Maryam Zomorrodi

Tam Pham

Farideh Z Bischoff

Tony J Pircher

Affiliations

Soon will be listed here.

Abstract

Enrichment of rare circulating tumor cells (CTCs) in blood is typically achieved using antibodies to epithelial cell adhesion molecule (EpCAM), with detection using cytokeratin (CK) antibodies. However, EpCAM and CK are not expressed in some tumors and can be downregulated during epithelial-to-mesenchymal transition. A micro-fluidic system, not limited to EpCAM or CK, was developed to use multiple antibodies for capture followed by detection using CEE-Enhanced (CE), a novel in situ staining method that fluorescently labels the capture antibodies bound to CTCs. Higher recovery of CTCs was demonstrated using antibody mixtures compared to anti-EpCAM. In addition, CK-positive breast cancer cells were found in 15 of 24 samples (63%; range 1-60 CTCs), while all samples contained additional CE-positive cells (range 1-41; median = 11; P = .02). Thus, antibody mixtures against a range of cell surface antigens enables capture of more CTCs than anti-EpCAM alone and CE staining enables the detection of CK-negative CTCs.

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References

de Bono J, Scher H, Montgomery R, Parker C, Miller M, Tissing H . Circulating tumor cells predict survival benefit from treatment in metastatic castration-resistant prostate cancer. Clin Cancer Res. 2008; 14(19):6302-9. DOI: 10.1158/1078-0432.CCR-08-0872. View

Kowalewska M, Nowak R, Chechlinska M . Implications of cancer-associated systemic inflammation for biomarker studies. Biochim Biophys Acta. 2010; 1806(2):163-71. DOI: 10.1016/j.bbcan.2010.06.002. View

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

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

Meng S, Tripathy D, Frenkel E, Shete S, Naftalis E, Huth J . Circulating tumor cells in patients with breast cancer dormancy. Clin Cancer Res. 2004; 10(24):8152-62. DOI: 10.1158/1078-0432.CCR-04-1110. View

Scher H, Jia X, de Bono J, Fleisher M, Pienta K, Raghavan D . Circulating tumour cells as prognostic markers in progressive, castration-resistant prostate cancer: a reanalysis of IMMC38 trial data. Lancet Oncol. 2009; 10(3):233-9. PMC: 2774131. DOI: 10.1016/S1470-2045(08)70340-1. View

Schroder C, Ruiters M, de Jong S, Tiebosch A, Wesseling J, Veenstra R . Detection of micrometastatic breast cancer by means of real time quantitative RT-PCR and immunostaining in perioperative blood samples and sentinel nodes. Int J Cancer. 2003; 106(4):611-618. DOI: 10.1002/ijc.11295. View

Pachmann K, Camara O, Kohlhase A, Rabenstein C, Kroll T, Runnebaum I . Assessing the efficacy of targeted therapy using circulating epithelial tumor cells (CETC): the example of SERM therapy monitoring as a unique tool to individualize therapy. J Cancer Res Clin Oncol. 2010; 137(5):821-8. PMC: 3074080. DOI: 10.1007/s00432-010-0942-4. View

Leversha M, Han J, Asgari Z, Danila D, Lin O, Gonzalez-Espinoza R . Fluorescence in situ hybridization analysis of circulating tumor cells in metastatic prostate cancer. Clin Cancer Res. 2009; 15(6):2091-7. PMC: 2875199. DOI: 10.1158/1078-0432.CCR-08-2036. View

10.

van der Gun B, Wasserkort R, Monami A, Jeltsch A, Rasko T, Slaska-Kiss K . Persistent downregulation of the pancarcinoma-associated epithelial cell adhesion molecule via active intranuclear methylation. Int J Cancer. 2008; 123(2):484-489. DOI: 10.1002/ijc.23476. View

11.

Coumans F, Doggen C, Attard G, de Bono J, Terstappen L . All circulating EpCAM+CK+CD45- objects predict overall survival in castration-resistant prostate cancer. Ann Oncol. 2010; 21(9):1851-1857. DOI: 10.1093/annonc/mdq030. View

12.

Tai K, Shiah S, Shieh Y, Kao Y, Chi C, Huang E . DNA methylation and histone modification regulate silencing of epithelial cell adhesion molecule for tumor invasion and progression. Oncogene. 2007; 26(27):3989-97. DOI: 10.1038/sj.onc.1210176. View

13.

Alix-Panabieres C, Vendrell J, Slijper M, Pelle O, Barbotte E, Mercier G . Full-length cytokeratin-19 is released by human tumor cells: a potential role in metastatic progression of breast cancer. Breast Cancer Res. 2009; 11(3):R39. PMC: 2716508. DOI: 10.1186/bcr2326. View

14.

Pestrin M, Bessi S, Galardi F, Truglia M, Biggeri A, Biagioni C . Correlation of HER2 status between primary tumors and corresponding circulating tumor cells in advanced breast cancer patients. Breast Cancer Res Treat. 2009; 118(3):523-30. DOI: 10.1007/s10549-009-0461-7. View

15.

Stott S, Hsu C, Tsukrov D, Yu M, Miyamoto D, Waltman B . Isolation of circulating tumor cells using a microvortex-generating herringbone-chip. Proc Natl Acad Sci U S A. 2010; 107(43):18392-7. PMC: 2972993. DOI: 10.1073/pnas.1012539107. View

16.

Marrinucci D, Bethel K, Lazar D, Fisher J, Huynh E, Clark P . Cytomorphology of circulating colorectal tumor cells:a small case series. J Oncol. 2010; 2010:861341. PMC: 2810476. DOI: 10.1155/2010/861341. View

17.

Shaffer D, Leversha M, Danila D, Lin O, Gonzalez-Espinoza R, Gu B . Circulating tumor cell analysis in patients with progressive castration-resistant prostate cancer. Clin Cancer Res. 2007; 13(7):2023-9. DOI: 10.1158/1078-0432.CCR-06-2701. View

18.

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

19.

Franzen B, Linder S, Alaiya A, Eriksson E, Uruy K, Hirano T . Analysis of polypeptide expression in benign and malignant human breast lesions: down-regulation of cytokeratins. Br J Cancer. 1996; 74(10):1632-8. PMC: 2074852. DOI: 10.1038/bjc.1996.600. View

20.

Nagrath S, Sequist L, Maheswaran S, Bell D, Irimia D, Ulkus L . Isolation of rare circulating tumour cells in cancer patients by microchip technology. Nature. 2007; 450(7173):1235-9. PMC: 3090667. DOI: 10.1038/nature06385. View