title: Evaluation of multiparameter flow cytometry for the detection of breast cancer tumor cells in blood samples.
abstract: We comparatively evaluated different cytokeratin (CK) reagents analyzed by flow cytometry (FCM) for the identification of the best combination of DNA/CK staining for detecting minimal numbers of breast cancer cells in peripheral blood (PB). In 59 primary breast cancer tumors, we comparatively analyzed the reactivity for up to 6 different anti-CK reagents using multiparameter FCM: anti-CK7, anti-CK20, anti-pan-CK, anti-CK8/CK18, anti-CK8, and anti-CK18. Afterward, dilutional experiments of Michigan Cancer Foundation (MCF)7 breast cancer cells in PB were performed, and the sensitivity of a DNA/CK18 staining was evaluated. Our results showed that anti-CK18 reagents were those providing the brightest and more sensitive staining for primary breast cancer tumor cells by FCM. Dilutional experiments of MCF cells in PB showed that the DNA/anti-CK18 double staining was highly specific for the identification of epithelial cells; its sensitivity ranged between 10(-6) and 10(-7) (detection of 1 tumor cell among 10(6) to 10(7) nucleated blood cells). Combined assessment of DNA cell contents and reactivity for CK18 by FCM is a sensitive method for the specific identification of breast cancer cells in PB.
authors.affiliation: Cytometry Service, University of Salamanca, Spain.
authors.forename: Ignacio
authors.initials: I
authors.lastname: Cruz
authors.forename: Juana
authors.initials: J
authors.lastname: Ciudad
authors.forename: Juan Jesús
authors.initials: JJ
authors.lastname: Cruz
authors.forename: Manuel
authors.initials: M
authors.lastname: Ramos
authors.forename: Alberto
authors.initials: A
authors.lastname: Gómez-Alonso
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Adansa
authors.forename: Cesar
authors.initials: C
authors.lastname: Rodríguez
authors.forename: Alberto
authors.initials: A
authors.lastname: Orfao
mesh_terms: D001943:Breast Neoplasms / Q000097:blood / Q000175:diagnosis*; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007633:Keratins / Q000032:analysis; D009360:Neoplastic Cells, Circulating
chemical_list: D004273:DNA, Neoplasm; D007633:Keratins