Flow cytometric double labeling technique for screening of multidrug resistance
✍ Scribed by Eric E. O. Gheuens; Dirk R. van Bockstaele; Maarten van der Keur; Hans J. Tanke; Allan T. van Oosterom; Ernst A. De Bruijn
- Publisher
- John Wiley and Sons
- Year
- 1991
- Tongue
- English
- Weight
- 854 KB
- Volume
- 12
- Category
- Article
- ISSN
- 0196-4763
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✦ Synopsis
We investigated the capabilities of flow cytometry in the analysis of a multidrug resistant (MDR) human ovarian cancer cell line 2780AD and its drug sensitive parental A2780. A functional assay using daunorubicin (DNR) as a fluorescent probe was combined with an immunofluorescence assay of P-glycoprotein (P-gp) using the monoclonal antibody MRK-16. Functionally MDR could be demonstrated by the lower DNR-content of MDR cells compared to DNR-content of drug sensitive cells. When incubation was performed with DNR in the presence of verapamil, DNR-content increased in the MDR cells. However the content of the A2780 cells was never attained. Differences in DNR-content were not related to differences in DNA-content. In experimental cell lines immunofluorescence data were inversely related with those of DNR-content: MDR cells had high levels of P-gp expression and low levels of DNR-content (and vice versa in drug sensitive cells). Both assays can be easily combined in a multiparametric flow cytometric procedure to evaluate both parameters simultaneously in the same cells. Analysis of clinical samples demonstrates the existence of aberrant subpopulations which would not be detected by using a single parameter assay.