Application of antibodies to intermediate filament proteins as tissue-specific probes in the flow cytometric analysis of complex tumors.
The flow cytometric (FCM) analysis of carcinomas is often hampered by the presence of stromal and inflammatory cells in the cell suspensions obtained from such neoplasms. Therefore, an FCM method was developed to distinguish epithelial from nonepithelial cells by using polyclonal and monoclonal antibodies to (cyto)keratins, the epithelial type of intermediate filament proteins. Using a model system of cultured bladder carcinoma (T24) and leukemia (MOLT-4) cells, we tested our hypothesis and procedures by labeling cell mixtures with these antibodies. After incubation with an appropriate intermediate filament antibody and propidium iodide staining, the DNA content and distribution of T24 cells could be analyzed separately from MOLT-4 cells. When applied to cell suspensions of endometrial carcinomas, bladder carcinomas and Grawitz tumors, only the epithelial (primarily carcinoma) cells were stained for cytokeratin; these cells could thus be analyzed separately from stromal, inflammatory and other nonepithelial cells. In this way, a more accurate FCM analysis of the malignant fraction within a tumor can be achieved.
D000230:Adenocarcinoma / Q000032:analysis / Q000473:pathology; D002295:Carcinoma, Transitional Cell / Q000473:pathology*; D002453:Cell Cycle; D002478:Cells, Cultured; D004273:DNA, Neoplasm / Q000032:analysis; D004848:Epithelium / Q000473:pathology; D005434:Flow Cytometry; D006801:Humans; D007158:Immunologic Techniques; D007381:Intermediate Filament Proteins / Q000032:analysis*; D007633:Keratins / Q000032:analysis / Q000276:immunology; D001749:Urinary Bladder Neoplasms / Q000473:pathology*; D014746:Vimentin / Q000032:analysis / Q000276:immunology
D004273:DNA, Neoplasm; D007381:Intermediate Filament Proteins; D014746:Vimentin; D007633:Keratins