Journal
AMERICAN JOURNAL OF CLINICAL PATHOLOGY
Volume 123, Issue 1, Pages 66-74Publisher
OXFORD UNIV PRESS INC
DOI: 10.1309/WP3QWKVJFYDHHXQD
Keywords
flow cytometry; minimal disease; cytokeratin; breast cancer; DNA ploidy
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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 106 to 107 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.
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