GENOTYPE-PHENOTYPE ANALYSES OF LOW-FREQUENCY TUMOR-CELLS USING COMPUTERIZED IMAGE MICROSCOPY

Citation
Vr. Litle et al., GENOTYPE-PHENOTYPE ANALYSES OF LOW-FREQUENCY TUMOR-CELLS USING COMPUTERIZED IMAGE MICROSCOPY, Cytometry, 23(4), 1996, pp. 344-349
Citations number
22
Categorie Soggetti
Cell Biology","Biochemical Research Methods
Journal title
ISSN journal
01964763
Volume
23
Issue
4
Year of publication
1996
Pages
344 - 349
Database
ISI
SICI code
0196-4763(1996)23:4<344:GAOLTU>2.0.ZU;2-Q
Abstract
Techniques to identify low frequency (i.e., 10(-4)-10(-5)) tumor cells in bone marrow and peripheral blood of cancer patients provide opport unities for early detection of disseminated disease and characterizati on of the properties of cells released from the tumor, Low frequency e pithelial cells in marrow and blood can be detected using immunophenot ypic markers directed against intracellular and/or cell surface antige ns. However, nonspecific phenotypic labeling may compromise the abilit y to discriminate tumor from nontumor cells, We describe optimization of slide-based approaches that facilitate identification and subsequen t molecular cytogenetic characterization of rare tumor cell population s in hematopoietic tissues, Colon tumor cells seeded in a hematopoieti c background provided a model system to optimize methodologies that ar e applicable to detection and quantification of micrometastases in cli nical specimens, Mixtures of cytogenetically aberrant epithelial cells and hematopoietic cells on slides were labeled with an anti-cytokerat in 20 (anti-CK20) antibody that recognizes >90% of colon adenocarcinom as, Computerized image analysis was used to record the location of the immunofluorescent cells on microscopic slides. Cells on slides were t hen hybridized using fluorescence in situ hybridization (FISH) with re peat-sequence DNA probes to detect aneusomies, Previously discriminate d epithelial cells were relocated for molecular cytogenetic characteri zation. Tumor cells present at frequencies approximating 5 x 10(-5) we re discriminated on the basis of immunofluorescence and cell size, A l ow frequency (4 x 10(-4)) population of normal hematopoietic cells als o labeled with anti-CK20 (data not shown). This strategy of sequential immunophenotyping and molecular cytogenetic analyses may be useful to discriminate tumor from nontumor cells in cancer patients with microm etastatic disease. (C) 1996 Wiley-Liss, Inc.