Abstract
Diagnostic cytology based on the examination of cells from body cavity fluids misses̃50% 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. 2was 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. ALDH1pos/CD44pos/CD24neg 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.
| Original language | English |
|---|---|
| Pages (from-to) | 132-143 |
| Number of pages | 12 |
| Journal | Cytometry Part A |
| Volume | 77 |
| Issue number | 2 |
| DOIs | |
| State | Published - Feb 1 2010 |
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Keywords
- Cells in body cavity fluids
- Flow cytometry
- Immunocytochemistry
- Tumor associated markers
ASJC Scopus subject areas
- Cell Biology
- Histology
- Pathology and Forensic Medicine
Cite this
Flow immunocytochemistry of marker expression in cells from body cavity fluids. / Krishan, Awtar; Ganjei-Azar, Parvin; Hamelik, Ronald; Sharma, Deepti; Reis, Isildinha; Nadji, Mehrdad.
In: Cytometry Part A, Vol. 77, No. 2, 01.02.2010, p. 132-143.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Flow immunocytochemistry of marker expression in cells from body cavity fluids
AU - Krishan, Awtar
AU - Ganjei-Azar, Parvin
AU - Hamelik, Ronald
AU - Sharma, Deepti
AU - Reis, Isildinha
AU - Nadji, Mehrdad
PY - 2010/2/1
Y1 - 2010/2/1
N2 - Diagnostic cytology based on the examination of cells from body cavity fluids misses̃50% 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. 2was 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. ALDH1pos/CD44pos/CD24neg 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.
AB - Diagnostic cytology based on the examination of cells from body cavity fluids misses̃50% 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. 2was 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. ALDH1pos/CD44pos/CD24neg 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.
KW - Cells in body cavity fluids
KW - Flow cytometry
KW - Immunocytochemistry
KW - Tumor associated markers
UR - http://www.scopus.com/inward/record.url?scp=77249113262&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77249113262&partnerID=8YFLogxK
U2 - 10.1002/cyto.a.20824
DO - 10.1002/cyto.a.20824
M3 - Article
C2 - 19899128
AN - SCOPUS:77249113262
VL - 77
SP - 132
EP - 143
JO - Cytometry. Part A : the journal of the International Society for Analytical Cytology
JF - Cytometry. Part A : the journal of the International Society for Analytical Cytology
SN - 1552-4922
IS - 2
ER -