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Flow immunocytochemistry of marker expression in cells from body cavity fluids
Author(s) -
Krishan Awtar,
GanjeiAzar Parvin,
Hamelik Ronald,
Sharma Deepti,
Reis Isildinha,
Nadji Mehrdad
Publication year - 2010
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20824
Subject(s) - flow cytometry , aneuploidy , cytology , biology , pathology , malignancy , cytometry , immunohistochemistry , immunocytochemistry , microbiology and biotechnology , medicine , gene , chromosome , genetics
Diagnostic cytology based on the examination of cells from body cavity fluids misses ∼50% of patients with a proven malignancy. In an earlier study, we used immunohistochemical detection of epithelial membrane antigen expression with flow cytometric detection of DNA aneuploidy to reduce the number of false negatives. In the present study, we have combined DNA flow cytometry with flow cytometric detection of marker expression to analyze cells from body cavity fluids. Seventy‐nine specimens of ascites and pleural fluids were analyzed by diagnostic cytology, DNA flow cytometry, and for the expression of the following markers: Ber‐EP4, progesterone (PR), MUC4, and thyroid transcription factor‐1 (TTF‐1). DNA index of equal to or greater than 1.2 was seen in 33/79 (41.7%) of the samples. Statistical analysis of 79 samples in which data from cytology, DNA aneuploidy, and expression of at least one of the markers was available showed that by combining data from positive marker expression with that of aneuploidy, the sensitivity was increased from 58.5 to 100%. In contrast, out of the 38 samples designated as non‐malignant by diagnostic cytology, nine had aneuploid DNA content and 16 of the diploid samples had a positive marker expression. Specificity was reduced from 74.7 to 31.6% due to the presence of aneuploidy and marker expression in these samples. ALDH1 pos /CD44 pos /CD24 neg expression has been reported to be associated with human breast tumor stem cells. Some of our samples had cells with this phenotype. Flow cytometry offers the advantage of rapid multiparametric analysis of DNA aneuploidy and marker expression in cells from body cavity fluids based on the analysis of a large number of cells without observer bias. By further developing the use of specific markers and aneuploidy, it may be possible to refine flow cytometric analysis for rapid detection of malignant cells in body cavity fluids. © 2009 International Society for Advancement of Cytometry