Abstract
To investigate the prognostic value of the molecular detection of circulating tumor cells (CTCs) using three markers [cytokeratin 19 (CK19), mammaglobin A (MGB1), and HER2] in early breast cancer. CK19mRNA+, MGB1mRNA+, and HER2mRNA+ cells were detected using real-time (CK19) and nested (MGB1 and HER2) reverse transcription-PCR in the peripheral blood of 175 women with stage I to III breast cancer before the initiation of adjuvant chemotherapy. The detection of CTCs was correlated with clinical outcome. In 10 patients, immunofluorescence staining experiments were done to investigate the coexpression of cytokeratin, MGB1, and HER2 in CTCs. CK19mRNA+, MGB1mRNA+, and HER2mRNA+ cells were detected in 41.1%, 8%, and 28.6% of the 175 patients, respectively. Patients had one of the following molecular profiles: CK19mRNA+/MGB1mRNA+/HER2mRNA+ (n = 8), CK19mRNA+/MGB1mRNA+/HER2mRNA- (n = 1), CK19mRNA+/MGB1mRNA-/HER2mRNA+ (n = 42), CK19mRNA+/MGB1mRNA-/HER2mRNA- (n = 21), CK19mRNA-/MGB1mRNA+/HER2mRNA- (n = 5), and CK19mRNA-/MGB1mRNA-/HER2mRNA- (n = 98). Double-immunofluorescence experiments confirmed the following CTC phenotypes: CK+/MGB1+, CK+/MGB1-, CK-/MGB1+, CK+/HER2+, CK+/HER2-, MGB1+/HER2-, and MGB1+/HER2+. In univariate analysis, the detection of CK19mRNA+, MGB1mRNA+, and HER2mRNA+ cells was associated with shorter disease-free survival (DFS; P < 0.001, P = 0.001, and P < 0.001, respectively), whereas the detection of CK19mRNA+ and MGB1mRNA+ cells was associated with worse overall survival (P = 0.044 and 0.034, respectively). In multivariate analysis, estrogen receptor-negative tumors and the detection of CK19mRNA+ and MGB1mRNA+ cells were independently associated with worse DFS. The detection of peripheral blood CK19mRNA+ and MGB1mRNA+ cells before adjuvant chemotherapy predicts poor DFS in women with early breast cancer.
Highlights
CK19mRNA+ and MGB1mRNA+ cells was associated with worse overall survival (P = 0.044 and 0.034, respectively)
We have previously shown that the detection of peripheral blood CK19mRNA+ cells before the initiation of adjuvant chemotherapy is an independent prognostic factor for shorter diseasefree survival (DFS) and overall survival (OS) [7, 8]
We report for the first time that, using a three-marker reverse transcriptionPCR (RT-PCR) analysis, the detection of CK19mRNA+ cells as well as the detection of MGB1mRNA+ cells were both independent prognostic factors for reduced DFS
Summary
Micrometastases, which are undetectable by the classic imaging and laboratory studies, can contribute to disease relapse; their identification in patients with early breast cancer may have substantial effect on determining prognosis and individualizing treatment strategies for these patients. For this purpose, epithelial cells have been identified in the bone marrow (disseminated tumor cells) or the peripheral blood [circulating tumor cells (CTCs)] of patients with stage I to III breast cancer using immunocytochemistry or molecular techniques (1 – 4). To increase the sensitivity of detection of CTCs as well as to define clinically relevant subpopulations of CTCs with aggressive biological behavior that could be used more precisely as surrogate markers for relapse, several approaches have been used: (a) the use of other markers apart from cytokeratins for detection of minimal residual disease [e.g., mucins, mammaglobin A (MGB1), maspin, carcinoembryonic antigen, HER2, and epidermal growth factor receptor; refs. 13 – 17], (b) the use of different multimarker RT-PCR assays [18, 19], and (c) the use of assays to distinguish between apoptotic and nonapoptotic or viable micrometastatic cells [20, 21]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
