Abstract
Multidrug-resistance (MDR), caused by overexpression of either P-glycoprotein (Pgp) or the multidrug-resistance associated protein (MRP), is characterised by a decreased cellular drug accumulation. One form of MDR is the sequestration of the drug inside cytoplasmic vesicles followed by an a exocytotic and/or efflux process. In some studies, increased intracellular glutathione (GSH) has been associated with MDR. In this study, we examined the effects of 7-chloro-4-nitrobenz-2-oxa-1,3-diazole or NBD (a H+-ATPase pump inhibitor) and buthionine sulphoximine or BSO (an inhibitor of GSH biosynthesis) on the subcellular distribution of daunorubicin or DNR in two leukemic homoharringtonine-resistant K562 cell lines, overexpressing MRP (K-H30) and Pgp (K-H300). DNR nuclear accumulation was carried out using microspectrofluorometry. Our results show that DNR nuclear accumulation and sensitivity of K-H30 cells were increased by NBD and BSO whereas in K-H300 cells, NBD and BSO were unable to increase the DNR nuclear accumulation and sensitivity of these cells. This study demonstrates clearly that even if vesicular sequestration can happen in cells overexpressing MRP and Pgp proteins, only the MRP protein is able to extrude the drug through intracellular vesicles and efflux. In addition, GSH plays an important part in the pathway of drug transport in cells overexpressing MRP. Data entrain also the notion of functional discrimination between the MDR and MRP phenotype.
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References
Endicott JA, Ling V, The biochemistry of P-glycoprotein-mediated multidrug resistance, Annu Rev Biochem, 1989, 58, 137–171.
Gottesman MM, Pastan I, Biochemistry of multidrug resistance mediated by the multidrug transporter, Annu Rev Biochem, 1993, 62, 385–427.
Zaman GJR, Flens MJ, Van Leusden MR, De Haas M, Mulder HS, Lankelma J, Pinedo HM, Scheper RJ, Baas F, Broxterman HJ, Borst P, The human multidrug resistance-associated protein MRP is a plasma membrane drug efflux pump, Proc Nat Acad Sci (Wash), 1994, 91, 8822–8826.
Leier I, Jedlitschky G, Buchholz U, Cole SPC, Deeley RG, Keppler D, The MRP gene encodes an ATP-dependent pump for leukotriene C4 and structurally related conjugates, J Biol Chem, 1994, 269, 27807–27810.
Zaman GJ, Lankelma J, Van Tellingen O, Role of glutathione in the export of compounds from cells by the multidrug resistance-associated protein, Proc Nat Acad Sci (Wash), 1995, 92, 7690–7694.
Cole SPC, Sparks KE, Fraser K, Loe DW, Grant CE, Wilson GM, Deely RG, Pharmacological characterization of multidrug resistant MRP-transfected human tumor cells, Cancer Res, 1994, 54, 5902–5910.
Almquist KC, Low DW, Hipfner DR, Characterization of the Mr 190,000 multidrug resistance protein (MRP) in drug-selected and transfected human tumor cells, Cancer Res 1995, 55, 102–110.
Meister A, Anderson ME, Glutathione, Annu Rev Biochem, 1983, 52, 711–760.
Ishikawa T, Ali-Osman F, Glutathione-associated cis-diamminedichloroplatinum (II) metabolism and ATP-dependent efflux from leukemia cells, J Biol Chem, 1993, 268, 20116–20125.
Ishikawa T, Wright C, Ishizuka H, GS-X pump is functionally overexpressed in cis-diamminedichloroplatinum (II)-resistant human leukemia HL60 cells and down-regulated by cell differentiation, J Biol Chem, 1994, 269, 29085–29093.
Ishikawa T, Bao J, Yamane Y, Akimaru K, Frindrich K, Wright C, Kuo MT, Coordinated induction of MRP/GS-X pump and γ-glutamylcysteine synthetase by heavy metals in human leukemia cells. J Biol Chem, 1996, 271, 14981–14988.
Tew KD, Glutathione-associated enzymes in anticancer drug resistance, Cancer Res, 1994, 54, 4313–4320.
Lutzky J, Astor MB, Taub RN, Baker MA, Bhalla K, Gervasoni JE, Rosado M, Stewart V, Krishana S, Hindenburg AA, Role of glutathione and dependent enzymes in anthracyclines-resistant HL60/AR cells, Cancer Res, 1989, 49, 4120–4125.
Morrow CS, Cowan KH, Glutathione S-transferases and drug resistance, Cancer Cells, 1990, 2, 15–22.
Commandeur JNM, Stijntjes GJ, Vermeulen NPE, Enzymes and transport systems involved in the formation and disposition of glutathione S-conjugates, Pharmacol Rev, 1995, 47, 271–330.
Zhou DC, Ramond S, Viguie F, Faussat AM, Zittoun R, Marie JP, Sequential emergence of MRP-and MDR1-gene over-expression as well as MDR1-gene translocation in homoharringtonine-selected K562 human leukemia cell lines, Int J Cancer, 1996, 65, 365–371.
Chomczynski P, Sacchi N, Single-Step-method of RNA isolation by acid guanidiniumthiocynatephenol-chloroform extration, Anal Biochem, 1987, 162, 156–159.
Gigli M, Doglia S, Millot JM, Valentini L, Manfait M, Quantitative study of doxorubicin in living cell nuclei by microspectrofluorometry, Biochem Biophys Acta, 1988, 950, 13–20.
Gigli M, Rasoanaivo TWD, Millot JM, Jeannesson P, Rizzo V, Jardillier JC, Arcamone F, Manfait M, Correlation between growth inhibition and intranuclear doxorubicin and 4′-iododoxorubicin quantitated in living K562 cells by microspectrofluorometry, Cancer Res 1989, 49, 560–564.
Millot JM, Rasoanaivo TWD, Morjani H, Manfait M, Role of the aclacinomycin A-doxorubicin associated inreversal of doxorubicin resistance in K562 tumour cells, Br J Cancer 1989, 69, 678–684.
Zhou DC, Zittoun R, Marie JP, Homoharringtonine: an effective new natural product in cancer chemotherapy, Bull Cancer, 1995, 82, 987–995.
Coly HM, Amos PR, Twentyman PR, Workman P, Examination by confocal fluorescence imaging microscopy of the subcetlular localisation of anthracyclines in parent and multidrug resistant cell lines, Br J Cancer, 1993, 67, 1316–1323.
Marquardt D, Center MS, Drug transport mechanism in HL60 cells isolated for resistance to adriamycin: evidence for nuclear drug accumulation in resistant cells, Cancer Res, 1992, 52, 3157–3163.
Schindler M, Grabski S, Hoff E, Simon SM, Defective pH regulation of acidic compartments in human breast cancer cells (MCF-7) is normalized in adriamycin-resistant cells (MCF-7 adr), Biochemistry, 1996, 35, 2811–2817.
Thiebault F, Currier SJ, Whitaker J, Haugland RF, Gottesman M, Pastan I, Willingham MC, Activity of the multidrug transporter results in alkalinisation of the cytosol: measurements of cytosolic pH by microinjection of a pH-sensitive dye, J Histochem Cytochem, 1990, 38, 685–690
Mannervik B, Danielson UH, Glutathione S-transferases-stucture and catalytic activity, CRC Crit Rev Biochem, 1988, 23, 283–337.
Ishikawa T, The ATP-dependent glutathione S-conjugate export pump, Trends Biochem Sci, 1992, 17, 463–468.
Hamilton TC, Winber MA, Louie KG, Batist G, Behrens BC, Tsuruo T, Grotzinger KR, Mckoy WM, Young RC, Ozols RF, Augmentation of adriamycin, melphalan and cisplatin cytotoxicity in drug-resistant and sensitive human ovarian carcinoma cell lines by buthionine sulfoximine mediated glutathione depletion, Biochem Pharmacol, 1985, 34, 2583–2586.
Mans DRA, Schuurhuis GJ, Treskes M, Lafleur MVM, Retel J, Pinedo HM, Lankelma J, Modulation by D, L-buthionine-S-R-sulphoximine of etoposide cytotoxicity on human non-small cell lung, ovarian and breast carcinoma cell lines, Eur J Cancer, 1992, 28A, 1447–1452.
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Benderra, Z., Morjani, H., Trussardi, A., Manfait, M. (1999). Evidence for Functional Discrimination Between Leukemic Cells Overexpressing Multidrug-Resistance Associated Protein and P-Glycoprotein. In: Kaspers, G.J.L., Pieters, R., Veerman, A.J.P. (eds) Drug Resistance in Leukemia and Lymphoma III. Advances in Experimental Medicine and Biology, vol 457. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-4811-9_17
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DOI: https://doi.org/10.1007/978-1-4615-4811-9_17
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