Identification of Circulating Tumour Cells in Early Stage Breast Cancer Patients Using Multi Marker Immunobead RT-PCR

Overview

Journal J Hematol Oncol

Publisher Biomed Central

Specialties Hematology
Oncology

Date 2009 Jun 9

PMID 19500345

Citations 9

Authors

Michael P Raynor

Sally-Anne Stephenson

Kenneth B Pittman

David C A Walsh

Michael A Henderson

Alexander Dobrovic

Affiliations

Soon will be listed here.

Abstract

Introduction: The ability to screen blood of early stage operable breast cancer patients for circulating tumour cells is of potential importance for identifying patients at risk of developing distant relapse. We present the results of a study of the efficacy of the immunobead RT-PCR method in identifying patients with circulating tumour cells.

Results: Immunomagnetic enrichment of circulating tumour cells followed by RT-PCR (immunobead RT-PCR) with a panel of five epithelial specific markers (ELF3, EPHB4, EGFR, MGB1 and TACSTD1) was used to screen for circulating tumour cells in the peripheral blood of 56 breast cancer patients. Twenty patients were positive for two or more RT-PCR markers, including seven patients who were node negative by conventional techniques. Significant increases in the frequency of marker positivity was seen in lymph node positive patients, in patients with high grade tumours and in patients with lymphovascular invasion. A strong trend towards improved disease free survival was seen for marker negative patients although it did not reach significance (p = 0.08).

Conclusion: Multi-marker immunobead RT-PCR analysis of peripheral blood is a robust assay that is capable of detecting circulating tumour cells in early stage breast cancer patients.

Citing Articles

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References

Kasimir-Bauer S, Oberhoff C, Schindler A, Seeber S . A summary of two clinical studies on tumor cell dissemination in primary and metastatic breast cancer: methods, prognostic significance and implication for alternative treatment protocols (Review). Int J Oncol. 2002; 20(5):1027-34. DOI: 10.3892/ijo.20.5.1027. View

Taback B, Chan A, Kuo C, Bostick P, Wang H, Giuliano A . Detection of occult metastatic breast cancer cells in blood by a multimolecular marker assay: correlation with clinical stage of disease. Cancer Res. 2001; 61(24):8845-50. View

Hayes D, Smerage J . Is there a role for circulating tumor cells in the management of breast cancer?. Clin Cancer Res. 2008; 14(12):3646-50. DOI: 10.1158/1078-0432.CCR-07-4481. View

Braun S, Vogl F, Naume B, Janni W, Osborne M, Coombes R . A pooled analysis of bone marrow micrometastasis in breast cancer. N Engl J Med. 2005; 353(8):793-802. DOI: 10.1056/NEJMoa050434. View

Bostick P, Huynh K, Sarantou T, Turner R, Qi K, Giuliano A . Detection of metastases in sentinel lymph nodes of breast cancer patients by multiple-marker RT-PCR. Int J Cancer. 1998; 79(6):645-51. DOI: 10.1002/(sici)1097-0215(19981218)79:6<645::aid-ijc16>3.0.co;2-r. View

Tymms M, Ng A, Thomas R, Schutte B, Zhou J, Eyre H . A novel epithelial-expressed ETS gene, ELF3: human and murine cDNA sequences, murine genomic organization, human mapping to 1q32.2 and expression in tissues and cancer. Oncogene. 1997; 15(20):2449-62. DOI: 10.1038/sj.onc.1201427. View

Salomon D, Brandt R, Ciardiello F, Normanno N . Epidermal growth factor-related peptides and their receptors in human malignancies. Crit Rev Oncol Hematol. 1995; 19(3):183-232. DOI: 10.1016/1040-8428(94)00144-i. View

Lloyd J, McIver C, Stephenson S, Hewett P, Rieger N, Hardingham J . Identification of early-stage colorectal cancer patients at risk of relapse post-resection by immunobead reverse transcription-PCR analysis of peritoneal lavage fluid for malignant cells. Clin Cancer Res. 2006; 12(2):417-23. DOI: 10.1158/1078-0432.CCR-05-1473. View

Zippelius A, Kufer P, Honold G, Kollermann M, Oberneder R, Schlimok G . Limitations of reverse-transcriptase polymerase chain reaction analyses for detection of micrometastatic epithelial cancer cells in bone marrow. J Clin Oncol. 1997; 15(7):2701-8. DOI: 10.1200/JCO.1997.15.7.2701. View

10.

Zach O, Lutz D . Mammaglobin remains a useful marker for the detection of breast cancer cells in peripheral blood. J Clin Oncol. 2005; 23(13):3160. DOI: 10.1200/JCO.2005.05.176. View

11.

de Graaf H, Maelandsmo G, Ruud P, Forus A, Oyjord T, Fodstad O . Ectopic expression of target genes may represent an inherent limitation of RT-PCR assays used for micrometastasis detection: studies on the epithelial glycoprotein gene EGP-2. Int J Cancer. 1997; 72(1):191-6. DOI: 10.1002/(sici)1097-0215(19970703)72:1<191::aid-ijc27>3.0.co;2-l. View

12.

Hardingham J, Kotasek D, Farmer B, Butler R, Mi J, Sage R . Immunobead-PCR: a technique for the detection of circulating tumor cells using immunomagnetic beads and the polymerase chain reaction. Cancer Res. 1993; 53(15):3455-8. View

13.

Kumar S, Singh J, Xia G, Krasnoperov V, Hassanieh L, Ley E . Receptor tyrosine kinase EphB4 is a survival factor in breast cancer. Am J Pathol. 2006; 169(1):279-93. PMC: 1698769. DOI: 10.2353/ajpath.2006.050889. View

14.

Noguchi S, Aihara T, Nakamori S, Motomura K, Inaji H, Imaoka S . The detection of breast carcinoma micrometastases in axillary lymph nodes by means of reverse transcriptase-polymerase chain reaction. Cancer. 1994; 74(5):1595-600. DOI: 10.1002/1097-0142(19940901)74:5<1595::aid-cncr2820740516>3.0.co;2-l. View

15.

Ignatiadis M, Kallergi G, Ntoulia M, Perraki M, Apostolaki S, Kafousi M . Prognostic value of the molecular detection of circulating tumor cells using a multimarker reverse transcription-PCR assay for cytokeratin 19, mammaglobin A, and HER2 in early breast cancer. Clin Cancer Res. 2008; 14(9):2593-600. DOI: 10.1158/1078-0432.CCR-07-4758. View

16.

Chang C, Scott G, Kuo W, Xiong X, Suzdaltseva Y, Park J . ESX: a structurally unique Ets overexpressed early during human breast tumorigenesis. Oncogene. 1997; 14(13):1617-22. DOI: 10.1038/sj.onc.1200978. View

17.

Branagan G, Hughes D, Jeffrey M, Crane-Robinson C, Perry P . Detection of micrometastases in lymph nodes from patients with breast cancer. Br J Surg. 2002; 89(1):86-9. DOI: 10.1046/j.0007-1323.2001.01970.x. View

18.

De Luca A, Pignata S, Casamassimi A, DAntonio A, Gridelli C, Rossi A . Detection of circulating tumor cells in carcinoma patients by a novel epidermal growth factor receptor reverse transcription-PCR assay. Clin Cancer Res. 2000; 6(4):1439-44. View

19.

Gradilone A, Gazzaniga P, Silvestri I, Gandini O, Trasatti L, Lauro S . Detection of CK19, CK20 and EGFR mRNAs in peripheral blood of carcinoma patients: correlation with clinical stage of disease. Oncol Rep. 2002; 10(1):217-22. View

20.

EATON M, Hardingham J, Kotasek D, Dobrovic A . Immunobead RT-PCR: a sensitive method for detection of circulating tumor cells. Biotechniques. 1997; 22(1):100-5. DOI: 10.2144/97221st01. View