Cancerous multi-drug resistance is reduced by Leptomycin B treatment in CCRF-CEM/Taxol cellsCancerous multi-drug resistance is reduced by Leptomycin B treatment in CCRF-CEM/Taxol cells

Abstract

Objectives: Multi-drug resistance (MDR) to chemotherapy remains a major obstacle to overcome in the successful treatment of patients with cancers. It was recently discovered that Leptomycin B (LMB) reduces the paclitaxel-induced MDR in CCRF-CEM/Taxol cells. However, the mechanism remains unclear. Here we sought to explore the mechanism of LMB to reduce the MDR induced by paclitaxel. Results: LMB has remarkable cytotoxic effects in both sensitive CCRF-CEM and resistant CCRF-CEM/Taxol cell lines. The paclitaxel-induced MDR was reduced by 0.013 μm of LMB. Lower concentration of LMB regulated cell cycle progress, in situ expressions of P-gp, MRP1, and LRP, expression of CRM1, and localization of P-gp and CRM1 in CCRF-CEM/taxol cells. Study Design: Cytotoxicity of LMB on cancerous cell lines was determined by MTT assay. Cell cycle progress and in situ expressions of P-gp, MRP1, and LRP were analyzed by flow cytometry. Expression of CRM1 in the cells was examined by Western blot. And co-localization between P-gp and CRM1 was determined by laser confocal microscopy. Conclusion: The paclitaxel-induced MDR of CCRFCEM/Taxol cells was reduced by lower concentration of LMB. The mechanisms might be related to decreasing in situ expression of drug transporter proteins, promoting cell cycle progress, and altering co-localization between P-gp and CRM1 in the resistant cells.

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Zhu, J. , Zhang, Y. and Guan, Y. (2012) Cancerous multi-drug resistance is reduced by Leptomycin B treatment in CCRF-CEM/Taxol cellsCancerous multi-drug resistance is reduced by Leptomycin B treatment in CCRF-CEM/Taxol cells. Health, 4, 845-855. doi: 10.4236/health.2012.410130.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] Higgins, C.F. (2007) Multiple molecular mechanisms for multi-drug resistance transporters. Nature, 446, 749-757. doi:10.1038/nature05630
[2] Lage, H. (2003) ABC-transporters: Implications on drug resistance from microorganisms to human cancers. International Journal of Antimicrobial Agents, 22, 188-199. doi:10.1016/S0924-8579(03)00203-6
[3] Scheffer, G.L., Wijngaard, P.L., Flens, M.J., Izquierdo, M.A., Slovak, M.L., Pinedo, H.M., et al. (1995) The drug resistance-related protein LRP is the human major vault protein. Nature Medicine, 6, 578-582. doi:10.1038/nm0695-578
[4] Engel, R., Valkov, N.I., Gump, J.L., Hazlehurst, L., Dalton, W.S. and Sullivan, D.M. (2004) The cytoplasmic trafficking of DNA topoisomerase IIa correlates with etoposide resistance in human myeloma cells. Experimental Cell Research, 295, 421-431. doi:10.1016/j.yexcr.2004.01.012
[5] Turner, J.G., Engel, R., Derderian, J.A., Jove, R. and Sullivan, D.M. (2004) Human topoisomerase IIa nuclear export is mediated by two CRM-1-dependent nuclear export signals. Journal of Cell Science, 117, 3061-3071. doi:10.1242/jcs.01147
[6] Longley, D.B. and Johnston, P.G. (2005) Molecular mechanisms of drug resistance. The Journal of Pathology, 205, 275-292. doi:10.1002/path.1706
[7] Turner, J.G., Dawson, J. and Sullivan, D.M. (2012) Nuclear export of proteins and drug resistance in cancer. Biochemical Pharmacology, 83, 1021-1032. doi:10.1016/j.bcp.2011.12.016
[8] Kim, J.W., Ho, W.J. and Wu, B.M. (2011) The role of the 3D environment in hypoxia-induced drug and apoptosis resistance. Anticancer Research, 31, 3237-3245.
[9] Zhu, J.W. and Guan, Y.B. (2010) Construction of cancerous multi-drug resistant cell model induced by paclitaxel from human leukemia CCRF-CEM cells. Chin J Pharmacol Toxicol, 24, 134-139.
[10] Fischer, U., Huber, J., Boelens, W.C., Mattaj, I.W. and Lührmann, R. (1995) The HIV-1 rev activation domain is a nuclear export signal that accesses an export pathway used by specific cellular RNAs. Cell, 82, 475-483. doi:10.1016/0092-8674(95)90436-0
[11] Kudo, N., Matsumori, N., Taoka, H., Fujiwara, D., Schreiner, E.P., Wolff, B., et al. (1999) Leptomycin B inactivates CRM1/exportin 1 by covalent modification at a cysteine residue in the central conserved region. Proceedings of the National Academy of Sciences, 96, 9112-9117.
[12] Gaubatz, S., Lees, J.A., Lindeman, G.L. and Livingston, D.M. (2001) E2F4 is exported from the nucleus in a CRM1-dependent Manner. Molecular and Cellular Biology, 21, 1384-1392. doi:10.1128/MCB.21.4.1384-1392.2001
[13] Julien, C., Coulombe, P. and Meloche, S. (2003) Nuclear export of ERK3 by a CRM1-dependent mechanism regulates its inhibitory action on cell cycle progression. The Journal of Biological Chemistry, 278, 42615-42624. doi:10.1074/jbc.M302724200
[14] Stommel, J.M., Marchenko, N.D., Jimenez, G.S., Moll, U.M., Hope, T.J. and Wahl, G.M. (1999) A leucine-rich nuclear export signal in the p53 tetramerization domain: Regulation of subcellular localization and p53 activity by NES masking. The EMBO Journal, 18, 1660-1672. doi:10.1093/emboj/18.6.1660
[15] Monte, M., Benetti, R., Collavin, L., Marchionni, L., Sal, G.D. and Schneider, C. (2004) hGTSE-1 expression stimulates cytoplasmic localization of p53. The Journal of Biological Chemistry, 279, 11744-11752. doi:10.1074/jbc.M311123200
[16] Grinberg, A.V., Hu, C.D. and Kerppola, T.K. (2004) Visualization of Myc/Max/Mad family dimers and the competition for dimerization in living cells. Molecular and Cellular Biology, 24, 4294-4308. doi:10.1128/MCB.24.10.4294-4308.2004
[17] Nishi, K., Yoshida, M., Fujiwara, D., Nishikawa, M., Horinouchi, S. and Beppu, T. (1994) Leptomycin B targets a regulatory cascade of crm1, a fission yeast nuclear protein, involved in control of higher order chromosome structure and gene expression. The Journal of Biological Chemistry, 269, 6320-6324.
[18] Vigneri, P. and Wang, J.Y. (2001) Induction of apoptosis in chronic myelogenous leukemia cells through nuclear entrapment of BCR-ABL tyrosine kinase. Nature Medicine, 7, 228-234. doi:10.1038/84683
[19] Shah, D., Naciri, M., Clee, P. and Mohamed, A.R. (2006) Nucleocounter: An efficient technique for the determination of cell number and viability in animal cell culture processes. Cytotechnology, 51, 39-44. doi:10.1007/s10616-006-9012-9
[20] Yamada, M., Ariga, T., Kawamura, N., Yamaguchi, K., Ohtsu, M., Nelson, D.L., et al. (2000) Determination of carrier status for the wiskott-aldrich syndrome by flow cytometric analysis of wiskott-aldrich syndrome protein expression in peripheral blood mononuclear cells. The Journal of Immunology, 165, 1119-1122.
[21] Meschini, S., Marra, M., Calcabrini, A., Monti, E., Gariboldi, M., Dolfini, E., et al. (2002) Role of the lung resistance-related protein (LRP) in the drug sensitivity of cultured tumor cells. Toxicol in Vitro, 16, 389-398. doi:10.1016/S0887-2333(02)00035-8
[22] Cortner, J. and Farnham, P.J. (1991) Cell cycle analysis of Krox-20, c-fos, and JE expression in proliferating NIH3T3 fibroblasts. Cell Growth & Differentiation, 2, 465-473.
[23] Jang, B.C., Paik, J.H. and Jeong, H.Y. (2004) Leptomycin B-induced apoptosis is mediated through caspase activetion and down-regulation of Mcl-1 and XIAP expression, but not through the generation of ROS in U937 leukemia cells. Biochemical Pharmacology, 68, 263-274. doi:10.1016/j.bcp.2004.03.007
[24] Laurencot, C.M., Scheffer, G.L. and Shoemaker, R.H. (1997) Incresed LRP mRNA expression is associated with the MDR phenotype in intrinsically resistant human cancer cell lines. International Journal of Cancer, 72, 1021-1026. doi:10.1002/(SICI)1097-0215(19970917)72:6<1021::AID-IJC17>3.0.CO;2-7
[25] Larsen, A.K., Escargueil, A.E. and Skladanowski, A. (2000) Resistance mechanisms associated with altered intracellular distribution of anticancer agents. Pharmacology & Therapeutics, 85, 217-229. doi:10.1016/S0163-7258(99)00073-X
[26] Krishna, R. and Mayer, L.D. (2000) Multidrug resistance (MDR) in cancer Mechanisms, reversal using modulators of MDR and the role of MDR modulators in influencing the pharmacokinetics of anticancer drugs. European Journal of Pharmaceutical Sciences, 11, 265-283. doi:10.1016/S0928-0987(00)00114-7
[27] Laurencot, C.M., Scheffer, G.L., Schrper, R.J. and Shoemarker, R.H. (1997) Increased LRP mRNA expression is associated with the MDR phenotype in intrinsically resistant human cancer cell lines. International Journal of Cancer, 72, 1021-1026. doi:10.1002/(SICI)1097-0215(19970917)72:6<1021::AID-IJC17>3.0.CO;2-7
[28] Ferrao, P., Sincock, P., Cole, S. and Ashman, L. (2001) Intracellular P-gp contributes to functional drug efflux and resistance in acute myeloid leukaemia. Leukemia Research, 25, 395-405. doi:10.1016/S0145-2126(00)00156-9
[29] Stade, K., Ford, C.S., Guthrie, C. and Weis, K. (1997) Exportin 1 (Crm1p) is an essential nuclear export factor. Cell, 90, 1041-1050. doi:10.1016/S0092-8674(00)80370-0
[30] Jang, B.C., Ursula, M.N., Paik, J.H., Claffey, K., Yoshida, M. and Hla, T. (2003) Leptomycin B, an inhibitor of the nuclear export receptor CRM1, inhibits COX-2 expression. The Journal of Biological Chemistry, 278, 2773-2776. doi:10.1074/jbc.C200620200

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