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Rapid suppression of multidrug resistance of leukemic cells by oxidative srtess

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Biochemistry (Moscow) Supplement Series A: Membrane and Cell Biology Aims and scope

Abstract

The effect of a short-time (1 h) oxidative stress on multidrug resistance (MDR) of murine leukemic P388VR cells has been investigated. We studied the production of reactive oxygen species (ROS) in cells depending on the composition of medium and the concentration of cells and hydrogen peroxide, as well as the effect of hydrogen peroxide on MDR of cells. MDR was determined from the transport of calcein acetoxymethyl ester out of the cells and from a change in cell sensitivity to vincristine. The amount of ROS arising in cells was determined using 2′,7′-dichlorodihydrofluorescein diacetate (DCFH2-DA). It was shown that the rate of ROS formation in cells decreases after the addition of serum to the medium and with an increase of the cell number. By the action of hydrogen peroxide, the amount of ROS increases directly with its concentration. Oxidative stress generated by 30–300 μM hydrogen peroxide decreases the MDR of the cells. The effect of hydrogen peroxide increases with the treatment duration and concentration of hydrogen peroxide. MDR determined by the criterion of the efflux of calcein ester from cells is completely suppressed after 1-h exposure to 300 μM hydrogen peroxide. At a concentration of hydrogen peroxide of 60 μM and treatment duration of 1 h, the sensitivity of P388VR cells to vincristine increases to reach the sensitivity of the wild-type P388 cells. Rapid (about 1 h) suppression of MDR is caused by inhibition of the activity of transport proteins. MDR decrease induced by oxidative stress can be used in therapy of tumors resistant to anticancer drugs.

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Abbreviations

DCF:

2′,7′-dichlorofluorescein

DCFH2 :

2′,7′-dichlorodihydrofluorescein

DCFH2-DA:

2′,7′-dichlorodihy-drofluorescein diacetate

MDR:

multidrug resistance

P-gp:

P-glycoprotein

ROS:

reactive oxygen species

References

  1. Stavrovskaya, A.A., Cellular Mechanisms of Multidrug Resistance of Tumor Cells, Biochemisrty, Moscow, 2000, vol. 65, no. 1, pp. 112–126.

    Google Scholar 

  2. Van der Bliek, A.M. and Borst, P., Multidrug Resistance, Adv. Cancer Res., 1989, vol. 52, pp. 165–203.

    Article  PubMed  Google Scholar 

  3. Leith, C., Multidrug Resistance in Leukemia, Curr. Opin. Hematol., 1998, vol. 5, no. 4, pp. 287–291.

    Article  CAS  PubMed  Google Scholar 

  4. List, A.F. and Spier, C.M., Multidrug Resistance in Acute Leukemia: a Conserved Physiologic Function, Leuk. Lymphoma, 1992, vol. 8, no. 1, pp. 9–14.

    Article  CAS  PubMed  Google Scholar 

  5. Chin, J.E., Soffir, R., Noonan, K.E., Choi, K. and Roninson I.B., Structure and Expression of the Human MDR (P-glycoprotein) Gene Family, Mol. Cell. Biol., 1989, vol. 9, no. 9, pp. 3808–3820.

    CAS  PubMed  Google Scholar 

  6. Gros, P., Ben Neriah, Y.B., Croop, J.M. and Housman, D.E., Isolation and Expression of a Complementary DNA that Confers Multidrug Resistance, Nature, 1986, vol. 323, no. 6090, pp. 728–731.

    Article  CAS  PubMed  Google Scholar 

  7. Gros, P., Raymond, M., Bell, J. and Housman, D., Cloning and Characterization of a Second Member of the Mouse mdr Gene Family, Mol. Cell. Biol., 1988, vol. 8, no. 7, pp. 2770–2778.

    CAS  PubMed  Google Scholar 

  8. Ziemann, C., Burklel, A., Kahl, G.F. and Hirsch-Ernst, K.I., Reactive Oxygen Species Participate in mdr1b mRNA and P-glycoprotein Overexpression in Primary Rat Hepatocyte Cultures, Carcinogenesis, 1999, vol. 20, no. 3, pp. 407–414.

    Article  CAS  PubMed  Google Scholar 

  9. Deng, L., Lin-Lee, Y-C., Claret, F-X. and Kuo, M.T., 2-acetometylaminofluorene up-Regulates Rat mdr 1b Expression through Generating Reactive Oxygen Species that Activate NF-κB Pathway, J. Biol. Chem., 2001, vol. 276, no. 1, pp. 413–420.

    Article  CAS  PubMed  Google Scholar 

  10. Felix, R.A. and Barrand, M.A., P-glycoprotein Expression in Rat Brain Endothelial Cells: Evidence for Regulation by Transient Oxidative Stress, J. Neurochem., 2002, vol. 80, no. 1, pp. 64–72.

    Article  CAS  PubMed  Google Scholar 

  11. Wartenberg, M., Fischer, K., Hescheler, J. and Sauer, H., Redox Regulation of P-glycoprotein-mediated Multidrug Resistance in Multicellular Prostate Tumor Spheroids, Int. J. Cancer, 2000, vol. 85, no. 2, pp. 267–274.

    CAS  PubMed  Google Scholar 

  12. Wartenberg, M., Ling, F.C., Muschen, M. and Klein, F., Regulation of the Multidrug Resistance Transporter P-glycoprotein in Multicellular Tumor Spheroids by Hypoxia-inducible Factor-1 and Reactive Oxygen Species, FASEB J., 2003, vol. 17, no. 3, pp. 503–505.

    CAS  PubMed  Google Scholar 

  13. Liang, B.C., Effects of Hypoxia on Drug Resistance Phenotype and Genotype in Human Glioma Cell Lines, J. Neurooncol., 1996, vol. 29, no. 2, pp. 149–155.

    Article  CAS  PubMed  Google Scholar 

  14. Ermakova, N.V., Shaposhnikova, V.V., Levitman, M.K., Kim, Y.A. and Korystov, Y.N., Alteration of Multidrug Resistance of Vincristine-resistant Lympholeukosis P388 Cells in the Case of Ascitic Tumor Growth, Biol. Membranes, 2004, vol. 21, no. 2, pp. 102–106.

    CAS  Google Scholar 

  15. Korystov, Y.N., Ermakova, N.V., Shaposhnikova, V.V., Levitman, M.K., Kim, Y.A. and Pushkin, S.Y., Perftoran Injection into P388 Ascitic Lympholeucosis Mouse Tumor Decreases the Number of Tumor Cells and their Multidrug Resistance, Biol. Membranes, 2006, vol. 23, no. 3, pp. 22–26.

    Google Scholar 

  16. Nagayama, J., Iino, M., Tada, Y., Kusaba, H., Kiue, A., Ohshima, K., Kuwano, M. and Wada M., Retrovirus Insertion and Transcriptional Activation of the Multidrug-resistance Gene in Leukemias Treated by a Chemotherapeutic Agent in Vivo, Blood, 2001, vol. 97, no. 3, pp. 759–766.

    Article  CAS  PubMed  Google Scholar 

  17. Zhou, J., Liu, M., Aneja, R., Chandra, R., Lage, H. and Joshi H-C., Reversal of P-glycoprotein-mediated Multidrug Resistance in Cancer Cells by the c-Jun NH2-Terminal Kinase, Cancer Research, 2006, vol. 66, no. 1, pp. 445–452.

    Article  CAS  PubMed  Google Scholar 

  18. Homolya, H., Hollo, Z., Germann, U.A., Pastan, I., Gottesman, M.M. and Sarkadi, B., Fluorescent Cellular Indicators Are Extruded by the Multidrug Resistance Protein, J. Biol. Chem., 1993, vol. 268, no. 29, pp. 21493–21496.

    CAS  PubMed  Google Scholar 

  19. Essodaigui, M., Broxterman, H.J. and Garnier-Suillerot, A., Kinetic Analysis of Calcein and Calcein-acetoxymethylester Efflux Mediated by the Multidrug Resistance Protein and P-glycoprotein, Biochemistry, 1998, vol. 37, no. 8, pp. 2243–2250.

    Article  CAS  PubMed  Google Scholar 

  20. Korystov, Y.N., Shaposhnikova, V.V., Korystova, A.F. and Emel’yanov, M.O., Detection of Reactive Oxygen Species Induced by Radiation in Cells Using the Dichlorofluorescein Assay, Radiat. Res., 2007, vol. 168, no. 2, pp. 226–232.

    Article  CAS  PubMed  Google Scholar 

  21. Korystov, Y.N., Emel’yanov, M.O., Korystova, A.F., Levitman, M.Kh. and Shaposhnikova, V.V., Determination of Reactive Oxygen and Nitrogen Species in Rat Aorta Using the Dichlorofluorescein Assay, Free Radic. Res., 2009, vol. 43, no. 2, pp. 149–155.

    Article  CAS  PubMed  Google Scholar 

  22. Fine, R.L., Chambers, T.C. and Sachs, C.W., P-glycoprotein, Multidrug Resistance and Protein Kinase C, The Oncologist, 1996, vol. 1, no. 4, pp. 261–268.

    CAS  PubMed  Google Scholar 

  23. Sharom, F.J., The P-glycoprotein Efflux Pump: How Does It Transport Drugs? J. Membrane Biol., 1997, vol. 160, no. 3, pp. 161–175.

    Article  CAS  Google Scholar 

  24. Castro, A.F., Horton, J.K., Vanoye, C.G. and Altenberg, G.A., Mechanism of Inhibition of P-glycoprotein-mediated Drug Transport by Protein Kinase C Blockers, Biochem. Pharmacol., 1999, vol. 58, no. 11, pp. 1723–1733.

    Article  CAS  PubMed  Google Scholar 

  25. Ido, A., Asao, T., Sakura, M., Inagaki, M., Saito, M. and Hidaka, H., An Inhibitor of Protein Kinase C, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine(H-7) Inhibits TPA-induced Reduction of Vincristine Uptake from P388 Murine Leukemic Cell, Leuk. Res., 1986, vol. 10, no. 9, pp. 1063–1069.

    Article  CAS  PubMed  Google Scholar 

  26. Schraufstater, I.U., Hyslop, P.A., Hinshaw, D.B., Spragg, R.G., Sklar, L.A. and Cochrane, C.G., Hydrogen Peroxide-induced Injury of Cells and Its Prevention by Inhibitors of Poly(ADP-ribose) Polymerase, Proc. Natl. Acad. Sci. USA, 1986, vol. 83. no. 13, pp. 4908–4912.

    Article  Google Scholar 

  27. Hyslop, P.A., Hinshawz, D.B., Halsey, W.A., Schraufstatter, I.U., Sauerhebery, R.D., Spraggj, R.G., Jackson, J.H. and Cochrane, C.G., Mechanisms of Oxidant-mediated Cell Injury. The Glycolytic and Mitochondrial Pathways of ADP Phosphorylation Are Major Intracellular Targets Inactivated by Hydrogen Peroxide, J. Biol. Chem. 1988, vol. 263. no. 4, pp. 1665–1675.

    Google Scholar 

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Authors and Affiliations

  1. Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino, Moscow oblast, 142290, Russia

    M. O. Emel’yanov, A. F. Korystova, L. N. Kublik, V. V. Shaposhnikova & Yu. N. Korystov

  2. Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow oblast, 142290, Russia

    Yu. A. Kim

Authors
  1. M. O. Emel’yanov
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  2. Yu. A. Kim
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  3. A. F. Korystova
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  4. L. N. Kublik
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  5. V. V. Shaposhnikova
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  6. Yu. N. Korystov
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Correspondence to Yu. N. Korystov.

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Original Russian Text © M.O. Emel’yanov, Yu.A. Kim, A.F. Korystova, L.N. Kublik, V. V. Shaposhnikova, Yu.N. Korystov, 2010, published in Biologicheskie Membrany, 2010, Vol. 27, No. 3, pp. 244–251.

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Emel’yanov, M.O., Kim, Y.A., Korystova, A.F. et al. Rapid suppression of multidrug resistance of leukemic cells by oxidative srtess. Biochem. Moscow Suppl. Ser. A 4, 212–219 (2010). https://doi.org/10.1134/S1990747810020133

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  • DOI: https://doi.org/10.1134/S1990747810020133

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