Abstract
Recent studies have identified a family of glycoproteins which moulate cellular transport of antibiotics, alkaloids and drugs used in cancer chemotherapy. By facilitating efflux of drugs from the intracellular domain, these proteins reduce cytotoxicity and thus confer drug resistance. With the availability of antibodies raised against these phenotypic markers of drug resistance, immunohistochemistry and flow cytometry has been used to study their distribution and expression in normal and tumor cells. As some of the drugs used in cancer chemotherapy and other dyes which are substrates for this efflux pump are fluorescent, laser flow cytometry can be used for rapid quantitation of cellular retention, efflux and heterogeneity in drug transport of a tumor cell population. This method can also be used to screen drugs which may block efflux of a chemotherpeutic drug and thus increase chemosensitivity of a drug resistant tumor. In the present report flow cytometric methods for the study of drug transport and its modulation in tumor cells are discussed.
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References
Almquist KC, Loe DW, Hipfner DR et al. (1995). Characterization of the Mr 190,000 multidrug resistance protein (Mrp) in drug-selected and transfected human tumor cells. Cancer Res 55: 102-110.
Deeley RG, Cole SP (1997). Function, evolution and structure of multidrug resistance protein (MRP). Semin Cancer Biol 8: 193-204.
Doyle LA, Yang W, Abruzzo LV et al. (1998). A multidrug resistance transporter from human MCF-7 breast cancer cells. Proc Natl Acad Sci USA 95: 15665-15670.
Frey T, Yue S, Haugland RP (1995). Dyes providing increased sensitivity in flow-cytometric dye-efflux assays for multidrug resistance. Cytometry 20: 218-227.
Ganapathi R, Grabowski D, Rouse W et al. (1981). Differential effect of the calmodulin inhibitor trifluoperazine on cellular accumulation, retention, and cytotoxicity of anthracyclines in doxorubicin (adriamycin)-resistant P388 mouse leukemia cells. Cancer Res 44: 5056-5061.
Hollo Z, Homolya L, Davis CW et al. (1994). Calcein accumulation as a fluorometric functional assay of the multidrug transporter. Biochim Biophys Acta 1191: 384-388.
Izquierdo M, Scheffer G, Flens M et al. (1996). Broad distribution of the multidrug resistance-related vault lung protein in normal human tissues and tumors. Am J Pathol 148: 877-887.
Juliano RL, Ling V (1976). A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. Biochim Biophys Acta 455: 152-162.
Krishan A (1987). Effect of drug efflux blockers on vital staining of cellular DNA with Hoechst 33341. Cytometry 8: 642-645.
Krishan A (2001). Monitoring of cellular resistance to cancer chemotherapy: drug retention and efflux. Methods Cell Biology 64: 193-209.
Krishan A, Fitz MC, Andritsch I (1997). Drug retention, efflux and resistance in tumor cells. Cytometry 29: 279-285.
Krishan A, Ganapathi R (1980). Laser flow cytometric studies on the intracellular fluorescence of anthracyclines. Cancer Res 40: 3895-3900.
Krishan A, Sauerteig A, Andritsch I, and Wellham L (1997). Flow cytometric analysis of the multiple drug resistance phenotype. Leukemia 11: 1138-1146.
Krishan A, Sauerteig A, Wellham L (1985). Flow cytometric studies on modulation of cellular adriamycin retention by phenothiazines. Cancer Res 45: 1046-1051.
Krishan A, Sridhar KS, Davila E et al. (1987). Patterns of anthracycline retention modulation in human tumor cells. Cytometry 8: 306-314.
Kunikane H, Zalupski MM, Ramachandran C et al. (1997). Flow cytometric analysis of P-glycoprotein expression and drug efflux in human soft tissue and bone sarcomas. Cytometry 30: 197-203.
Lampidis T, Munck J, Krishan A et al. (1985). Reversal of resistance to rhodamine 123 in adriamycin-resistant Friend leukemia cells. Cancer Res 45: 2626-2631.
Leith CP, Kopecky KJ, Chen IM (1999). Frequency and clinical significance of the expression of multi drug resistance proteins MDR1/P-gp, MRP1, and LRP in acute myeloid leukemia. A Southwest Oncology Group study. Blood 94: 1086-1099.
Lelong EH, Guzikowski AP, Haugland RP et al. (1991). Fluorescent verapamil derivative for monitoring activity of the multidrug transporter. Mol Pharmacol 40: 490-494.
Ling V (1992). P-glycoprotein and resistance to anti-cancer drugs. Cancer 69: 2603-2609.
Nair S, Singh SV, Samy TSA et al. (1990). Anthracycline resistance in murine leukemic P388 cells: Role of drug efflux and glutathione related enzymes. Biochemical Pharmacol 39: 723-728.
Ross DD, Gao Y, Yang W et al. (1997). The 95-kilodalton membrane glycoprotein overexpressed in novel multidrug resistant breast cancer cells is NCA, the nonspecific cross-reacting antigen of carcinoembryonic antigen. Cancer Res 57: 5460-5464.
Scheffer GL, Wijngaard PLJ, Flens MJ et al. (1995). The drug resistance related protein LRP is the human major vault protein. Nat Med 1: 578-582.
Sikic BI, Fisher GA, Lum BL et al. (1997). Modulation and prevention of multidrug resistance by inhibitors of P-glycoprotein. Cancer Chemother Pharmacol 40: Suppl S13-S19.
Slater LM, Sweet P, Stupecky M et al. (1986). Cyclosporin A corrects daunorubicin resistance in Ehrlich ascites carcinoma. Br J Cancer 54: 235-238.
Tsuruo T, Iida H, Tsukagoshi S et al. (1981). Overcoming of vincristine resistance in P388 leukemia in vivoand in vitrothrough enhanced cytotoxicity of vincristine and vinblastine by verapamil. Cancer Res 41: 1967-1972.
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Krishan, A. Flow cytometric monitoring of drug resistance in human tumor cells. Methods Cell Sci 24, 55–60 (2002). https://doi.org/10.1023/A:1024185612997
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DOI: https://doi.org/10.1023/A:1024185612997