Detection of EpCAM positive and negative circulating tumor cells in metastatic breast cancer patients.
BACKGROUND
Immunomagnetic EpCAM based methods are used to enrich circulating tumor cells (CTCs) in metastatic breast cancer (mBC) patients. EpCAM negative CTCs may be missed. We addressed the question of the reliability of an EpCAM dependent assay to enrich CTCs.


METHODS
To elucidate this issue, our study has been designed to assess two different CTC enrichment technologies (i) in EpCAM positive (+) and EpCAM negative cell lines and (ii) in mBC patients in dependency on their respective EpCAM expression. These two technologies encompass one anti-EpCAM immunomagnetic enrichment technology, MACS HEA MicroBeads(®) (MACS), and one EpCAM independent density centrifugation method, OncoQuick(®) plus (OQ+). Furthermore, the coherence between EpCAM expression in the primary tumor tissue of mBC patients and the CTC detection rates in the corresponding patients is analyzed.


RESULTS
(i) MACS recovered significantly more EpCAM (+) than EpCAM (-) tumor cells (p < 0.001) in spiked blood samples. With OQ+ no significantly different recovery rates between EpCAM (+) and EpCAM (-) tumor cells (p = 0.796) were detected. (ii) In mBC patients MACS yielded a significantly higher (p = 0.024) detection rate of EpCAM (+) CTCs. No statistically significant difference (p = 0.070) was found concerning the EpCAM status-based detection rate of CTCs by OQ+. (iii) CTC detection rates are independent of the primary tumors' EpCAM expression.


CONCLUSIONS
EpCAM (-) CTCs can not be detected by immunomagnetic EpCAM dependent enrichment methods. EpCAM independent enrichment technologies seem to be superior to detect the entire CTC population. Evaluation of CTCs as prognostic marker should compromise EpCAM (+) and (-) subpopulations.
Applied Cancer Research - Institution for Translational Research Vienna/CEADDP, Austria. robert.koenigsberg@gmx.at
Robert
R
Königsberg
Eva
E
Obermayr
Giovanna
G
Bises
Georg
G
Pfeiler
Margit
M
Gneist
Fritz
F
Wrba
Maria
M
de Santis
Robert
R
Zeillinger
Marcus
M
Hudec
Christian
C
Dittrich
D000328:Adult; D000368:Aged; D000951:Antigens, Neoplasm / Q000097:blood / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000378:metabolism; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology*; D002277:Carcinoma / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology*; D016022:Case-Control Studies; D015815:Cell Adhesion Molecules / Q000097:blood / Q000378:metabolism*; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods; D000071858:Epithelial Cell Adhesion Molecule; D005260:Female; D006801:Humans; D008875:Middle Aged; D009362:Neoplasm Metastasis; D009360:Neoplastic Cells, Circulating / Q000378:metabolism / Q000473:pathology*; D011379:Prognosis; D012680:Sensitivity and Specificity
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule