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Detection and Characterization of Viable Circulating Tumor Cells

from Molecular Medicine Congress 2013

Since current assays cannot distinguish between apoptotic and viable DTCs/CTCs, it is now possible to apply a novel ELISPOT assay (designated ‘EPISPOT’) that detects proteins secreted/released/shed from single epithelial cancer cells. Cells are cultured for a short time on a membrane coated with antibodies that capture the secreted/released/shed proteins which are subsequently detected by secondary antibodies labeled with fluorochromes. In breast cancer, we measured the release of cytokeratin-19 (CK19) and mucin-1 (MUC1) and demonstrated that many patients harbored viable DTCs, even in patients with apparently localized tumors (stage M0 : 54%). Preliminary clinical data showed that patients with DTC-releasing CK19 have an unfavorable outcome. We also studied CTCs or CK19-releasing cells (CK19-RC) in the peripheral blood of M1 breast cancer patients and showed that patients with CK19-RC had a worse clinical outcome. In prostate cancer, we used prostate-specific antigen (PSA) secretion as the marker and found that a significant fraction of CTCs secreted fibroblast growth factor-2 (FGF-2), a known stem cell growth factor. More recently, in colon cancer, a considerable portion of viable CTCs detectable by the Epispot assay is trapped in the liver as the first filter organ in colon cancer patients. The enumeration of CK19-SC by the CK19-Epispot assay revealed viable CTCs in 65.9% and 55.4% (p=0.04) patients in mesenteric and peripheral blood, respectively, whereas CellSearch detected CTCs in 55.9% and 29.0% (p=0.0046) patients. In mesenteric blood, the number of CTC was significantly higher than in the peripheral blood. Our clinical data showed that localized colon cancer patients with a high level of CTCs have an unfavorable outcome (n=60). In conclusion, the EPISPOT assay offers a new opportunity to detect and characterize viable DTCs/CTCs in cancer patients and it can be extended to a multi-parameter analysis revealing a CTC/DTC protein fingerprint.

about 8 years ago

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