Flow cytometry DNA analysis on tumor cell subpopulation of human tumor specimens by exclusion of lymphohemopoietic cells.
We describe a new flow cytometry procedure in which DNA analyses can be obtained selectively on pure, freshly obtained tumor cell subpopulations of human tumor specimens. This procedure is based on exclusion from analysis of the contaminating lymphohemopoietic cells mixed with tumor cells in tumor specimens. This exclusion is made possible by labeling all lymphohemopoietic cells with an antibody to HLe-1 (HLE), which is present on all lymphohemopoietic cells but on no other cells, and by gating against these labeled cells when analyzing for DNA. For the model system, a 1:1 mixture of normal human peripheral blood leukocytes and either of two human cancer cell lines, HEp-2 and MCF-7, normal leukocyte contamination can be reduced to 3.1% while retaining 94.7% of tumor cells for DNA analysis. Four examples of human tumor samples, two cases each of malignant effusions and lymph node metastases, were analyzed with this procedure. The results clearly indicate that this new method will improve ploidy analysis/aneuploidy detection and will make it possible to obtain more accurate cell-cycle analyses of tumor cells than have previously been possible. This new procedure will contribute to clinical and biological studies involving DNA of human tumors.
Park
C H
CH
Department of Medicine, University of Kansas Medical Center, College of Health Sciences and Hospital, Kansas City 66103.
Lee
S H
SH
Stephens
R L
RL
Smith
T K
TK
Park
M H
MH
D000230:Adenocarcinoma / Q000032:analysis; D000911:Antibodies, Monoclonal / Q000276:immunology; D001773:Blood Cells / Q000032:analysis*; D001943:Breast Neoplasms / Q000032:analysis; D002294:Carcinoma, Squamous Cell / Q000032:analysis; D004273:DNA, Neoplasm / Q000032:analysis*; D005434:Flow Cytometry; D006801:Humans; D008207:Lymphatic Metastasis; D008214:Lymphocytes / Q000032:analysis; D010996:Pleural Effusion
D000911:Antibodies, Monoclonal; D004273:DNA, Neoplasm