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Perturbation of the lipid phase of a membrane is not involved in the modulation of MRP1 transport activity by flavonoids

Abstract

The expression of transmembrane transporter multidrug resistance-associated protein 1 (MRP1) confers the multidrug-resistant phenotype (MDR) on cancer cells. Since the activity of the other MDR transporter, P-glycoprotein, is sensitive to membrane perturbation, we aimed to check whether the changes in lipid bilayer properties induced by flavones (apigenin, acacetin) and flavonols (morin, myricetin) were related to their MRP1 inhibitory activity. All the flavonoids inhibited the efflux of MRP1 fluorescent substrate from human erythrocytes and breast cancer cells. Morin was also found to stimulate the ATPase activity of erythrocyte ghosts. All flavonoids intercalated into phosphatidylcholine bilayers as judged by differential scanning calorimetry and fluorescence spectroscopy with the use of two carbocyanine dyes. The model of an intramembrane localization for flavones and flavonols was proposed. No clear relationship was found between the membrane-perturbing activity of flavonoids and their potency to inhibit MRP1. We concluded that mechanisms other than perturbation of the lipid phase of membranes were responsible for inhibition of MRP1 by the flavonoids.

Abbreviations

BCECF-AM:

2′,7′-bis-(3-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester

BCRP:

breast cancer resistance protein

ΔH:

enthalpy change of the main phospholipid phase transition

DiOC2(3):

3,3′-diethyloxacarbocyanine iodide

DiOC6(3):

3,3′-dihexyloxacarbocyanine iodide

DMPC:

1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine

DPPC:

1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine

EYPC:

egg yolk L-α-phosphatidylcholine

MDR:

multidrug resistance

MRP1:

multidrug resistance-associated protein 1

NBD:

nucleotide binding domain

P-gp:

P-glycoprotein

TM :

temperature of the main phospholipid phase transition

References

  1. Hipfner, D.R., Deeley, R.G. and Cole, S.P. Structural, mechanistic and clinical aspects of MRP1. Biochim. Biophys. Acta 1461 (1999) 359–376.

    PubMed  Article  CAS  Google Scholar 

  2. Keppler, D., Leier, I. and Jedlitschky, G. Transport of glutathione conjugates and glucuronides by the multidrug resistance proteins MRP1 and MRP2. Biol. Chem. 378 (1997) 787–791.

    PubMed  CAS  Google Scholar 

  3. Kamp, D. and Haest, C.W. Evidence for a role of the multidrug resistance protein (MRP) in the outward translocation of NBD-phospholipids in the erythrocyte membrane. Biochim. Biophys. Acta 1372 (1998) 91–101.

    PubMed  Article  CAS  Google Scholar 

  4. Dekkers, D.W., Comfurius, P., van Gool, R.G., Bevers, E.M. and Zwaal, R.F. Multidrug resistance protein 1 regulates lipid asymmetry in erythrocyte membranes. Biochem. J. 350 (2000) 531–535.

    PubMed  Article  CAS  Google Scholar 

  5. Raggers, R.J., van Helvoort, A., Evers, R. and van Meer, G.R.J. The human multidrug resistance protein MRP1 translocates sphingolipid analogs across the plasma membrane. J. Cell. Sci. 112 (1999) 415–422.

    PubMed  CAS  Google Scholar 

  6. Harborne, J.B. and Williams, C.A. Advances in flavonoid research since 1992. Phytochemistry 55 (2000) 481–504.

    PubMed  Article  CAS  Google Scholar 

  7. Dixon, R.A. and Ferreira, D. Genistein. Phytochemistry 60 (2000) 205–211.

    Article  Google Scholar 

  8. Heim, K.E., Tagliaferro, A.R. and Bobilya, D.J. Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. J. Nutr. Biochem. 13 (2002) 572–584.

    PubMed  Article  CAS  Google Scholar 

  9. Leslie, E.M., Mao, Q., Oleschuk, C.J., Deeley, R.G. and Cole, S.P. Modulation of multidrug resistance protein 1 (MRP1/ABCC1) transport and ATPase activities by interaction with dietary flavonoids. Mol. Pharmacol. 59 (2001) 1171–1180.

    PubMed  CAS  Google Scholar 

  10. Nguyen, H., Zhang, S. and Morris, M.E. Effect of flavonoids on MRP1-mediated transport in Panc-1 cells. J. Pharm. Sci. 92 (2003) 250–257.

    PubMed  Article  CAS  Google Scholar 

  11. Bobrowska-Hagerstrand, M., Wrobel, A., Mrowczynska, L., Soderstrom, T., Shirataki, Y., Motohashi, N., Molnar, J., Michalak, K. and Hagerstrand, H. Flavonoids as inhibitors of MRP1-like efflux activity in human erythrocytes. A structure-activity relationship study. Oncol. Res. 13 (2003) 463–469.

    PubMed  Google Scholar 

  12. Lania-Pietrzak, B., Michalak, K., Hendrich, A.B., Mosiadz, D., Grynkiewicz, G., Motohashi, N. and Shirataki, Y. Modulation of MRP1 protein transport by plant, and synthetically modified flavonoids. Life Sci. 77 (2005) 1879–1891.

    PubMed  Article  CAS  Google Scholar 

  13. Critchfield, J.W., Welsh, C.J., Phang, J.M. and Yeh, G.C. Modulation of adriamycin accumulation and efflux by flavonoids in HCT-15 colon cells. Activation of P-glycoprotein as a putative mechanism. Biochem. Pharmacol. 48 (1994 1437–1445.

    PubMed  Article  CAS  Google Scholar 

  14. Zhang, S. and Morris, M.E. Effects of the flavonoids biochanin A, morin, phloretin, and silymarin on P-glycoprotein-mediated transport. J. Pharmacol. Exp. Ther. 304 (2003) 1258–1267.

    PubMed  Article  CAS  Google Scholar 

  15. Imai, Y., Tsukahara, S., Asada, S. and Sugimoto, Y. Phytoestrogens/flavonoids reverse breast cancer resistance protein/ABCG2-mediated multidrug resistance. Cancer Res. 64 (2004) 4346–4352.

    PubMed  Article  CAS  Google Scholar 

  16. Ferte, J. Analysis of the tangled relationships between P-glycoprotein-mediated multidrug resistance and the lipid phase of the cell membrane. Eur. J. Biochem. 267 (2000) 277–294.

    PubMed  Article  CAS  Google Scholar 

  17. Hendrich, A.B. and Michalak, K. Lipids as a target for drugs modulating multidrug resistance of cancer cells. Curr. Drug Targets 4 (2003) 23–30.

    PubMed  Article  CAS  Google Scholar 

  18. Zordan-Nudo, T., Ling, V., Liu, Z. and Georges, E. Effects of nonionic detergents on P-glycoprotein drug binding and reversal of multidrug resistance. Cancer Res. 53 (1993) 5994–6000.

    PubMed  CAS  Google Scholar 

  19. Ford, J.M. and Hait, W.N. Pharmacology of drugs that alter multidrug resistance in cancer. Pharmacol. Rev. 42 (1990) 155–199.

    PubMed  CAS  Google Scholar 

  20. Wadkins, R.M. and Houghton, P.J. The role of drug-lipid interactions in the biological activity of modulators of multi-drug resistance. Biochim. Biophys. Acta 1153 (1993) 225–236.

    PubMed  Article  CAS  Google Scholar 

  21. Drori, S., Eytan, G.D. and Assaraf, Y.G. Potentiation of anticancer-drug cytotoxicity by multidrug-resistance chemosensitizers involves alterations in membrane fluidity leading to increased membrane permeability. Eur. J. Biochem. 228 (1995) 1020–1029.

    PubMed  Article  CAS  Google Scholar 

  22. Pajeva, I.K., Wiese, M., Cordes, H.P. and Seydel, J.K. Membrane interactions of some catamphiphilic drugs and relation to their multidrug-resistance-reversing ability. J. Cancer Res. Clin. Oncol. 122 (1996) 27–40.

    PubMed  Article  CAS  Google Scholar 

  23. Hendrich, A.B., Wesolowska, O., Motohashi, N., Molnar, J. and Michalak, K. New phenothiazine-type multidrug resistance modifiers: anti-MDR activity versus membrane perturbing potency. Biochem. Biophys. Res. Commun. 304 (2003) 260–265.

    PubMed  Article  CAS  Google Scholar 

  24. Bobrowska-Hagerstrand, M., Wrobel, A., Mrowczynska, L., Soderstrom, T. and Hagerstrand, H. Modulation of MRP1-like efflux activity in human erythrocytes caused by membrane perturbing agents. Mol. Membr. Biol. 20 (2003) 255–259.

    PubMed  Article  CAS  Google Scholar 

  25. Alqawi, O. and Georges, E. The multidrug resistance protein ABCC1 drug-binding domains show selective sensitivity to mild detergents. Biochem. Biophys. Res. Commun. 303 (2003) 1135–1141.

    PubMed  Article  CAS  Google Scholar 

  26. Marbeuf-Gueye, C., Stierle, V., Sudwan, P., Salerno, M. and Garnier-Suillerot, A. Perturbation of membrane microdomains in GLC4 multidrug-resistant lung cancer cells-modification of ABCC1 (MRP1) localization and functionality. FEBS J. 274 (2007) 1470–1480.

    PubMed  Article  CAS  Google Scholar 

  27. Klappe, K., Hinrichs, J.W., Kroesen, B.J., Sietsma, H. and Kok, J.W. MRP1 and glucosylceramide are coordinately over expressed and enriched in rafts during multidrug resistance acquisition in colon cancer cells. Int. J. Cancer 110 (2004) 511–522.

    PubMed  Article  CAS  Google Scholar 

  28. Hendrich, A.B. Flavonoid-membrane interactions: possible consequences for biological effects of some polyphenolic compounds. Acta Pharmacol. Sin. 27 (2006) 27–40.

    PubMed  Article  CAS  Google Scholar 

  29. Trompier, D., Baubichon-Cortay, H., Chang, X.B., Maitrejean, M., Barron, D., Riordon, J.R. and Di Pietro, A. Multiple flavonoid-binding sites within multidrug resistance protein MRP1. Cell. Mol. Life Sci. 60 (2003) 1–14.

    Article  Google Scholar 

  30. Wu, C.P., Calcagno, A.M., Hladky, S.B., Ambudkar, S.V. and Barrand, M.A. Modulatory effects of plant phenols on human multidrug-resistance proteins 1, 4 and 5 (ABCC1, 4 and 5). FEBS J. 272 (2005) 4725–4740.

    PubMed  Article  CAS  Google Scholar 

  31. Steck, T.L. and Kant, J.A. Preparation of impermeable ghosts and inside-out vesicles from human erythrocyte membranes. Methods Enzymol. 31 (1973) 172–180.

    Article  Google Scholar 

  32. Lowry, O.H., Roserbrough, N.J., Farr, A.L. and Randall, R.J. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193 (1951) 265–275.

    PubMed  CAS  Google Scholar 

  33. Rychlik, B., Balcerczyk, A., Klimczak, A. and Bartosz, G. The role of multidrug resistance protein 1 (MRP1) in transport of fluorescent anions across the human erythrocyte membrane. J. Membr. Biol. 193 (2003) 79–90.

    PubMed  Article  CAS  Google Scholar 

  34. Van Veldhoven, P.P. and Mannaerts, G.P. Inorganic and organic phosphate measurements in the nanomolar range. Anal. Biochem. 161 (1987) 45–48.

    PubMed  Article  Google Scholar 

  35. Ferreira, M.J., Gyemant, N., Madureira, A.M., Tanaka, M., Koos, K., Didziapetris, R. and Molnar, J. The effects of jatrophane derivatives on the reversion of MDR1-and MRP-mediated multidrug resistance in the MDAMB-231 (HTB-26) cell line. Anticancer Res. 25 (2005) 4173–4178.

    PubMed  CAS  Google Scholar 

  36. Matsumoto, Y., Takano, H., Kunishio, K., Nagao, S. and Fojo, T. Expression of drug resistance genes in VP-16 and mAMSA-selected human carcinoma cells. Jpn. J. Cancer Res. 92 (2001) 778–784.

    PubMed  CAS  Google Scholar 

  37. Pastan, I., Gottesman, M.M., Ueda, K., Lovelace, E., Rutherford, A.V. and Willingham, M.C. A retrovirus carrying an MDR1 cDNA confers multidrug resistance and polarized expression of P-glycoprotein in MDCK cells. Proc. Natl. Acad. Sci. USA 85 (1988) 4486–4490.

    PubMed  Article  CAS  Google Scholar 

  38. Ghose, A.K., Pritchett, A. and Crippen, G.M. Atomic physicochemical parameters for three dimensional structure activity relationships III: modelling hydrophobic interactions. J. Comput. Chem. 9 (1988) 80–90.

    Article  CAS  Google Scholar 

  39. Pulaski, L., Jedlitschky, G., Leier, I., Buchholz, U. and Keppler, D. Identification of the multidrug-resistance protein (MRP) as the glutathione-S-conjugate export pump of erythrocytes. Eur. J. Biochem. 241 (1996) 644–648.

    PubMed  Article  CAS  Google Scholar 

  40. Klokouzas, A., Wu, C.P., van Veen, H.W., Barrand, M.A. and Hladky, S.B. cGMP and glutathione-conjugate transport in human erythrocytes. Eur. J. Biochem. 270 (2003) 3696–3708.

    PubMed  Article  CAS  Google Scholar 

  41. Jedlitschky, G., Burchell, B. and Keppler, D. The multidrug resistance protein 5 functions as an ATP-dependent export pump for cyclic nucleotides. J. Biol. Chem. 275 (2000) 30069–30074.

    PubMed  Article  CAS  Google Scholar 

  42. Wesolowska, O., Mosiadz, D., Motohashi, N., Kawase, M. and Michalak, K. Phenothiazine maleates stimulate MRP1 transport activity in human erythrocytes. Biochim. Biophys. Acta 1720 (2005) 52–58.

    PubMed  Article  CAS  Google Scholar 

  43. McAleer, M.A., Breen, M.A., White, N.L. and Matthews, N. pABC11 (also known as MOAT-C and MRP5), a member of the ABC family of proteins, has anion transporter activity but does not confer multidrug resistance when over-expressed in human embryonic kidney 293 cells. J. Biol. Chem. 274 (1999) 23541–23548.

    PubMed  Article  CAS  Google Scholar 

  44. Klokouzas, A., Barrand, M.A. and Hladky, S.B. Effects of clotrimazole on transport mediated by multidrug resistance associated protein 1 (MRP1) in human erythrocytes and tumour cells. FEBS Lett. 268 (2001) 6569–6577.

    CAS  Google Scholar 

  45. Rychlik, B., Pulaski, L., Sokal, A., Soszynski, M. and Bartosz, G. Transport of organic anions by multidrug resistance-associated protein in the erythrocyte. Acta Biochim. Pol. 47 (2000) 763–772.

    PubMed  CAS  Google Scholar 

  46. Bobrowska-Hagerstrand, M., Wrobel, A., Rychlik, B., Bartosz, G., Soderstrom, T., Shirataki, Y., Motohashi, N., Molnar, J., Michalak, K. and Hagerstrand, H. Monitoring of MRP-like activity in human erythrocytes: inhibitory effect of isoflavones. Blood Cells Mol. Dis. 27 (2001) 894–900.

    PubMed  Article  CAS  Google Scholar 

  47. van Zanden, J.J., Wortelboer, H.M., Bijlsma, S., Punt, A., Usta, M., Bladeren, P.J., Rietjens, I.M. and Cnubben, N.H. Quantitative structure activity relationship studies on the flavonoid mediated inhibition of multidrug resistance proteins 1 and 2. Biochem. Pharmacol. 69 (2005) 699–708.

    PubMed  Article  CAS  Google Scholar 

  48. van Zanden, J.J., de Mul, A., Wortelboer, H.M., Usta, M., van Bladeren, P.J., Rietjens, I.M. and Cnubben, N.H. Reversal of in vitro cellular MRP1 and MRP2 mediated vincristine resistance by the flavonoid myricetin. Biochem. Pharmacol. 69 (2005) 1657–1665.

    PubMed  Article  CAS  Google Scholar 

  49. Phang, J.M., Poore, C.M., Lopaczynska, J. and Yeh, G.C. Flavonol-stimulated efflux of 7,12-dimethylbenz(a)anthracene in multidrug-resistant breast cancer cells. Cancer Res. 53 (1993) 5977–5981.

    PubMed  CAS  Google Scholar 

  50. Castro, A.F. and Altenberg, G.A. Inhibition of drug transport by genistein in multidrug-resistant cells expressing P-glycoprotein. Biochem. Pharmacol. 53 (1997) 89–93.

    PubMed  Article  Google Scholar 

  51. Boumendjel, A., Di Pietro, A., Dumontet, C. and Barron, D. Recent advances in the discovery of flavonoids and analogs with high-affinity binding to P-glycoprotein responsible for cancer cell multidrug resistance. Med. Res. Rev. 22 (2002) 512–529.

    PubMed  Article  CAS  Google Scholar 

  52. Ikegawa, T., Ohtani, H., Koyabu, N., Juichi, M., Iwase, Y., Ito, C., Furukawa, H., Naito, M., Tsuruo, T. and Sawada, Y. Inhibition of P-glycoprotein by flavonoid derivatives in adriamycin-resistant human myelogenous leukemia (K562/ADM) cells. Cancer Lett. 177 (2002) 89–93.

    PubMed  Article  CAS  Google Scholar 

  53. Conseil, G., Baubichon-Cortay, H., Dayan, G., Jault, J.M., Barron, D. and Di Pietro, A. Flavonoids: a class of modulators with bifunctional interactions at vicinal ATP-and steroid-binding sites on mouse P-glycoprotein. Proc. Natl. Acad. Sci. USA 95 (1998) 9831–9836.

    PubMed  Article  CAS  Google Scholar 

  54. Hooijberg, J.H., Pinedo, H.M., Vrasdonk, C., Priebe, W., Lankelma, J. and Broxterman, H.J. The effect of glutathione on the ATPase activity of MRP1 in its natural membranes. FEBS Lett. 469 (2000) 47–51.

    PubMed  Article  CAS  Google Scholar 

  55. Di Pietro, A., Conseil, G., Perez-Victoria, J.M., Dayan, G., Baubichon-Cortay, H., Trompier, D., Steinfels, E., Jault, J.M., de Wet, H., Maitrejean, M., Comte, G., Boumendjel, A., Mariotte, A.M., Dumontet, C., McIntosh, D.B., Goffeau, A., Castanys, S., Gamarro, F. and Barron, D. Modulation by flavonoids of cell multidrug resistance mediated by P-glycoprotein and related ABC transporters. Cell. Mol. Life Sci. 59 (2002) 307–322.

    PubMed  Article  Google Scholar 

  56. Mao, Q., Leslie, E.M., Deeley, R.G. and Cole, S.P. ATPase activity of purified and reconstituted multidrug resistance protein MRP1 from drug-selected H69AR cells. Biochim. Biophys. Acta 1461 (1999) 69–82.

    PubMed  Article  CAS  Google Scholar 

  57. Chang, X.B., Hou, Y.X. and Riordan, J.R. ATPase activity of purified multidrug resistance-associated protein. J. Biol. Chem. 272 (1997) 30962–30968.

    PubMed  Article  CAS  Google Scholar 

  58. Hooijberg, J.H., Broxterman, H.J., Heijn, M., Fles, D.L., Lankelma, J. and Pinedo, H.M. Modulation by (iso)flavonoids of the ATPase activity of the multidrug resistance protein. FEBS Lett. 413 (1997) 344–348.

    PubMed  Article  CAS  Google Scholar 

  59. Hooijberg, J.H., Broxterman, H.J., Scheffer, G.L., Vrasdonk, C., Heijn, M., de Jong, M.C., Scheper, R.J., Lankelma, J. and Pinedo, H.M. Potent interaction of flavopiridol with MRP1. Br. J. Cancer 81 (1999) 269–276.

    PubMed  Article  CAS  Google Scholar 

  60. Jain, M.K. and Wu, N.M. Effect of small molecules on the dipalmitoyl lecithin liposomal bilayer: III. Phase transition in lipid bilayer. J. Membr. Biol. 34 (1977) 157–201.

    Article  CAS  Google Scholar 

  61. Saija, A., Bonina, F., Trombetta, D., Tomaino, A., Montenegro, L., Smeriglio, P. and Castelli, F. Flavonoid-biomembrane interactions: A calorimetric study on dipalmitoylphosphatidylcholine vesicles. Int. J. Pharmaceutics 124 (1995) 1–8.

    Article  CAS  Google Scholar 

  62. Hendrich, A.B., Malon, R., Pola, A., Shirataki, Y., Motohashi, N. and Michalak, K. Differential interaction of Sophora isoflavonoids with lipid bilayers. Eur. J. Pharm. Sci. 16 (2002) 201–208.

    PubMed  Article  CAS  Google Scholar 

  63. Ollila, F., Haing, K., Vuorela, P., Vuorela, H. and Slotte, J.P. Characterization of flavonoid-biomembrane interactions. Arch. Biochem. Biophys. 399 (2002) 103–108.

    PubMed  Article  CAS  Google Scholar 

  64. Bhowmik, B.B., Basu, S. and Ray, D. Photophysical studies of 3,3′ dioctadecyloxacarbocyanine dye in model biological membranes and different solvents. Chem. Phys. Lipids 109 (2001) 175–183.

    PubMed  Article  CAS  Google Scholar 

  65. Sidorowicz, A., Pola, A. and Dobryszycki, P. Spectral properties of 3,3′-diethyloxadicarbocyanine included in phospholipid liposomes. J. Photochem. Photobiol. B 38 (1997) 94–97.

    Article  CAS  Google Scholar 

  66. Sidorowicz, A., Mora, C., Jablonka, S., Pola, A., Modrzycka, T., Mosiadz, D. and Michalak, K. Spectral properties of two betaine-type cyanine dyes in surfactant micelles and in the presence of phospholipids. J. Mol. Struct. 744–747 (2005) 711–716.

    Article  CAS  Google Scholar 

  67. Krishna, M.M. and Periasamy, N. Location and orientation of DODCI in lipid bilayer membranes: effects of lipid chain length and unsaturation. Biochim. Biophys. Acta 1461 (1999) 58–68.

    PubMed  Article  CAS  Google Scholar 

  68. Lopes, S. and Castanho, M.A. Does aliphatic chain length influence carbocyanines’ orientation in supported lipid multilayers? J. Fluorescence 14 (2004) 281–287.

    Article  CAS  Google Scholar 

  69. Scheidt, H.A., Pampel, A., Nissler, L., Gebhardt, R. and Huster, D. Investigation of the membrane localization and distribution of flavonoids by high-resolution magic angle spinning NMR spectroscopy. Biochim. Biophys. Acta 1663 (2004) 97–107.

    PubMed  Article  CAS  Google Scholar 

  70. Siarheyeva, A., Lopez, J.J. and Glaubitz, C. Localization of multidrug transporter substrates within model membranes. Biochemistry 45 (2006) 6203–6211.

    PubMed  Article  CAS  Google Scholar 

  71. Yu, J., Cheng, Y., Xie, L. and Zhang, R. Effects of genistein and daidzein on membrane characteristics of HCT cells. Nutr. Cancer 33 (1999) 100–104.

    PubMed  CAS  Article  Google Scholar 

  72. Oteiza, P.I., Erlejman, A.G., Verstraeten, S.V., Keen, C.L. and Fraga, C.G. Flavonoid-membrane interactions: a protective role of flavonoids at the membrane surface? Clin. Develop. Immunol. 12 (2005) 19–25.

    Article  CAS  Google Scholar 

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Wesołowska, O., Hendrich, A.B., Łania-Pietrzak, B. et al. Perturbation of the lipid phase of a membrane is not involved in the modulation of MRP1 transport activity by flavonoids. Cell Mol Biol Lett 14, 199–221 (2009). https://doi.org/10.2478/s11658-008-0044-3

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Key words

  • Flavonoids
  • Multidrug resistance-associated protein 1 (MRP1)
  • Lipid bilayer
  • Carbocyanine dyes