This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Stathopoulou, A. Articles by Lianidou, E. S. Search for Related Content PubMed PubMed Citation Articles by Stathopoulou, A. Articles by Lianidou, E. S.

Real-Time Quantification of CK-19 mRNA-Positive Cells in Peripheral Blood of Breast Cancer Patients Using the Lightcycler System

Laboratory of Analytical Chemistry, University of Athens, Athens [A. S., A. G., E. S. L.]; Department of Medical Oncology, University General Hospital of Heraklion, Heraklion [A. S., M. P., S. A., D. M., V. G.]; and Medical Oncology Unit, "Helena Venizelou" Hospital, Athens [N. M.], Greece

ABSTRACT

Purpose: The purpose of this research was to develop a quantitative real-time reverse transcription-PCR (RT-PCR) for CK19-mRNA and evaluate its clinical potential for the molecular detection of occult carcinoma cells in the peripheral blood of breast cancer patients.

Experimental Design: The method is based on real-time monitoring during PCR of fluorescently labeled specific hybridization probes for CK19-mRNA. The breast cancer cell line MCF-7 was used for the development and analytical evaluation of the assay. We analyzed blood samples from 89 healthy blood donors, 77 patients with early breast cancer (stage I-II) postoperatively, and 47 patients with previously untreated metastatic disease (stage IV) before and after chemotherapy. All of the samples were also analyzed by nested RT-PCR.

Results: The method is highly sensitive and specific, because only 2 of 89 (2.2%) of the healthy control subjects had detectable CK19-mRNA+ cells. In 77 patients with early breast cancer, CK19-mRNA+ cells were detected in 24 (31.2%) before and 5 (6.5%) after adjuvant chemotherapy, and their levels differed significantly (P < 0.001, Wilcoxon test). In 47 patients with verified metastases 19 (40.4%) and 20 (42.6%) were found positive before and after chemotherapy, and no significant difference in CK19-mRNA+ cell levels was observed (P = 0.96, Wilcoxon test). Results obtained by the proposed real-time RT-PCR method correlated well with those obtained for the same samples by nested RT-PCR [concordance in 312 of 337 (92.6%); P = 0.69, McNemar test].

Conclusions: The developed method is highly sensitive and specific, and can be used for high-throughput continuous monitoring and quantification of circulating epithelial cells in the peripheral blood of breast cancer patients.

This article has been cited by other articles:

				D. F. Hayes and J. Smerage Is There a Role for Circulating Tumor Cells in the Management of Breast Cancer? Clin. Cancer Res., June 15, 2008; 14(12): 3646 - 3650. [Abstract] [Full Text] [PDF]

				M. Ignatiadis, G. Kallergi, M. Ntoulia, M. Perraki, S. Apostolaki, M. Kafousi, G. Chlouverakis, E. Stathopoulos, E. Lianidou, V. Georgoulias, et al. 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., May 1, 2008; 14(9): 2593 - 2600. [Abstract] [Full Text] [PDF]

				M. Ignatiadis, N. Xenidis, M. Perraki, S. Apostolaki, E. Politaki, M. Kafousi, E. N. Stathopoulos, A. Stathopoulou, E. Lianidou, G. Chlouverakis, et al. Different Prognostic Value of Cytokeratin-19 mRNA Positive Circulating Tumor Cells According to Estrogen Receptor and HER2 Status in Early-Stage Breast Cancer J. Clin. Oncol., November 20, 2007; 25(33): 5194 - 5202. [Abstract] [Full Text] [PDF]

				L. Harris, H. Fritsche, R. Mennel, L. Norton, P. Ravdin, S. Taube, M. R. Somerfield, D. F. Hayes, and R. C. Bast Jr American Society of Clinical Oncology 2007 Update of Recommendations for the Use of Tumor Markers in Breast Cancer J. Clin. Oncol., November 20, 2007; 25(33): 5287 - 5312. [Abstract] [Full Text] [PDF]

				N. Xenidis, V. Markos, S. Apostolaki, M. Perraki, A. Pallis, G. Sfakiotaki, D. Papadatos-Pastos, L. Kalmanti, M. Kafousi, E. Stathopoulos, et al. Clinical relevance of circulating CK-19 mRNA-positive cells detected during the adjuvant tamoxifen treatment in patients with early breast cancer Ann. Onc., October 1, 2007; 18(10): 1623 - 1631. [Abstract] [Full Text] [PDF]

				N. Xenidis, M. Perraki, M. Kafousi, S. Apostolaki, I. Bolonaki, A. Stathopoulou, K. Kalbakis, N. Androulakis, C. Kouroussis, T. Pallis, et al. Predictive and Prognostic Value of Peripheral Blood Cytokeratin-19 mRNA-Positive Cells Detected by Real-Time Polymerase Chain Reaction in Node-Negative Breast Cancer Patients J. Clin. Oncol., August 10, 2006; 24(23): 3756 - 3762. [Abstract] [Full Text] [PDF]

				K. Koyanagi, S. J. O'Day, R. Gonzalez, K. Lewis, W. A. Robinson, T. T. Amatruda, C. Kuo, H.-J. Wang, R. Milford, D. L. Morton, et al. Microphthalmia Transcription Factor as a Molecular Marker for Circulating Tumor Cell Detection in Blood of Melanoma Patients Clin. Cancer Res., February 15, 2006; 12(4): 1137 - 1143. [Abstract] [Full Text] [PDF]

				G. Giribaldi, S. Procida, D. Ulliers, F. Mannu, R. Volpatto, G. Mandili, L. Fanchini, O. Bertetto, G. Fronda, L. Simula, et al. Specific Detection of Cytokeratin 20-Positive Cells in Blood of Colorectal and Breast Cancer Patients by a High Sensitivity Real-Time Reverse Transcriptase-Polymerase Chain Reaction Method J. Mol. Diagn., February 1, 2006; 8(1): 105 - 112. [Abstract] [Full Text] [PDF]

				K. Koyanagi, S. J. O'Day, R. Gonzalez, K. Lewis, W. A. Robinson, T. T. Amatruda, H.-J. Wang, R. M. Elashoff, H. Takeuchi, N. Umetani, et al. Serial Monitoring of Circulating Melanoma Cells During Neoadjuvant Biochemotherapy for Stage III Melanoma: Outcome Prediction in a Multicenter Trial J. Clin. Oncol., November 1, 2005; 23(31): 8057 - 8064. [Abstract] [Full Text] [PDF]

				K. Koyanagi, C. Kuo, T. Nakagawa, T. Mori, H. Ueno, A. R. Lorico Jr, H.-J. Wang, E. Hseuh, S. J. O'Day, and D. S.B. Hoon Multimarker Quantitative Real-Time PCR Detection of Circulating Melanoma Cells in Peripheral Blood: Relation to Disease Stage in Melanoma Patients Clin. Chem., June 1, 2005; 51(6): 981 - 988. [Abstract] [Full Text] [PDF]

				V. Bozionellou, D. Mavroudis, M. Perraki, S. Papadopoulos, S. Apostolaki, E. Stathopoulos, A. Stathopoulou, E. Lianidou, and V. Georgoulias Trastuzumab Administration Can Effectively Target Chemotherapy-Resistant Cytokeratin-19 Messenger RNA-Positive Tumor Cells in the Peripheral Blood and Bone Marrow of Patients With Breast Cancer Clin. Cancer Res., December 15, 2004; 10(24): 8185 - 8194. [Abstract] [Full Text] [PDF]

				A M Gilbey, D Burnett, R E Coleman, and I Holen The detection of circulating breast cancer cells in blood J. Clin. Pathol., September 1, 2004; 57(9): 903 - 911. [Abstract] [Full Text] [PDF]