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Flow Cytometric Characterization of Antifolate Resistance in Cultured Mammalian Cells Using Fluoresceinated Methotrexate and Daunorubicin¹

Departments of Biological Sciences [S. W. S., Y. G. A., R. T. S.] and Medicine (Oncology) [A. M.], Stanford University, Stanford, California 94305

We have studied antifolate-resistant rodent cell lines with respect to dihydrofolate reductase gene expression and expression of the "classic" multidrug resistance phenotype by flow cytometry. Using a series of antifolate-resistant and colchicine-resistant Chinese hamster ovary cell lines obtained by single-step and stepwise selection protocols, we show that viable cell staining with fluoresceinated methotrexate and daunorubicin correlates well with drug resistance and expression of dihydrofolate reductase protein and the "classic" MDR phenotype in these cell lines. We show that flow cytometric analysis makes it possible to rapidly assess the potentially complex drug resistance phenotype(s) of cells selected with hydrophilic and lipophilic antifolates.

¹ Y. G. A. is supported by a fellowship from the John D. and Catherine T. MacArthur Foundation. A. M. is supported by Robert Wood Johnson Foundation (grant 14546) and funds donated in the memory of Frances Eiger. This work was supported by grant NIH CA16318 to R. T. S.

² To whom requests for reprints should be addressed, at Department of Biological Sciences, Stanford University, Stanford, CA 94305.

Received 10/ 5/89. Revised 5/10/90.

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