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Modulation of the atypical multidrug-resistant phenotype by a hammerhead ribozyme directed against the ABC transporter BCRP/MXR/ABCG2

Cancer Gene Therapy volume 9, pages 579–586 (2002)Cite this article

Abstract

The phenomenon of multidrug resistance (MDR) in human cancers is one of the major causes of failure of chemotherapy. A recently identified new member of the superfamily of ATP-binding cassette transporters, breast cancer resistance protein (BCRP), was demonstrated to confer an atypical multidrug-resistant phenotype to tumor cells. To overcome the BCRP-mediated drug resistance, a specific anti-BCRP hammerhead ribozyme was introduced into the human gastric carcinoma cell line, EPG85-257RNOV, exhibiting an atypical MDR phenotype. By this approach, the expression levels of the targeted BCRP-encoding mRNA and the BCRP transport protein were decreased to the low constitutive expression level that was observed in highly drug-sensitive parental gastric carcinoma cells. In addition, in the anti-BCRP ribozyme-treated cells, the cellular drug accumulation was dramatically increased to the level measured in drug-sensitive cells. These effects were accompanied by an extensive reversal of the drug-resistant phenotype of more than 80%. Because additional mechanisms contribute to the multimodal-mediated MDR phenotype exhibited by this gastric carcinoma cell line, the data suggest that the BCRP-mediated contingent to the drug resistance was overcome nearly completely. Moreover, the data indicate that ribozyme-based gene therapy may be clinically applicable in preventing and reversing BCRP-mediated atypical MDR.

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References

  1. Gottesman MM, Fojo T, Bates SE . Multidrug resistance in cancer: role of ATP-dependent transporters Nat Rev Cancer 2002 2: 48–58

    Article  CAS  PubMed  Google Scholar 

  2. Borst P, Evers R, Kool M, Wijnholds J . A family of drug transporters: the multidrug resistance–associated proteins J Natl Cancer Inst 2000 92: 1295–1302

    Article  CAS  PubMed  Google Scholar 

  3. Lage H, Dietel M . Effect of the breast cancer resistance protein on atypical multidrug resistance Lancet Oncol 2000 1: 169–175

    Article  CAS  PubMed  Google Scholar 

  4. Ross DD, Yang W, Abruzzo LV et al. Atypical multidrug resistance: breast cancer resistance protein messenger RNA expression in mitoxantrone-selected cell lines J Natl Cancer Inst 1999 91: 429–433

    Article  CAS  PubMed  Google Scholar 

  5. Doyle LA, Yang W, Abruzzo LV et al. A multidrug resistance transporter from human MCF-7 breast cancer cells Proc Natl Acad Sci USA 1998 95: 15665–15670

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Volm M . Multidrug resistance and its reversal Anticancer Res 1998 18: 2905–2918

    CAS  PubMed  Google Scholar 

  7. Rabindran SK, Ross DD, Doyle LA, Yang W, Greenberger LM . Fumitremorgin C reverses multidrug resistance in cells transfected with the breast cancer resistance protein Cancer Res 2000 60: 47–50

    CAS  PubMed  Google Scholar 

  8. van Loevezijn A, Allen JD, Schinkel AH, Koomen GJ . Inhibition of BCRP-mediated drug efflux by fumitremorgin-type indolyl diketopiperazines Bioorg Med Chem Lett 2001 11: 29–32

    Article  CAS  PubMed  Google Scholar 

  9. de Bruin M, Miyake K, Litman T, Robey R, Bates SE . Reversal of resistance by GF120918 in cell lines expressing the ABC half-transporter, MXR Cancer Lett 1999 146: 117–126

    Article  CAS  PubMed  Google Scholar 

  10. Erlichman C, Boerner SA, Hallgren CG et al. The HER tyrosine kinase inhibitor CI1033 enhances cytotoxicity of 7-ethyl-10-hydroxycamptothecin and topotecan by inhibiting breast cancer resistance protein-mediated drug efflux Cancer Res 2001 61: 739–748

    CAS  PubMed  Google Scholar 

  11. Maliepaard M, van Gastelen MA, Tohgo A et al. Circumvention of breast cancer resistance protein (BCRP)–mediated resistance to camptothecins in vitro using non-substrate drugs or the BCRP inhibitor GF120918 Clin Cancer Res 2001 7: 935–941

    CAS  PubMed  Google Scholar 

  12. Perego P, De Cesare M, De Isabella P et al. A novel 7–modified camptothecin analog overcomes breast cancer resistance protein-associated resistance in a mitoxantrone-selected colon carcinoma cell line Cancer Res 2001 61: 6034–6037

    CAS  PubMed  Google Scholar 

  13. Kobayashi H, Dorai T, Holland JF, Ohnuma T . Reversal of drug sensitivity in multidrug-resistant tumor cells by an MDR1(PGY1) ribozyme Cancer Res 1994 54: 1271–1275

    CAS  PubMed  Google Scholar 

  14. Holm PS, Scanlon KJ, Dietel M . Reversion of multidrug resistance in the P-glycoprotein–positive human pancreatic cell line (EPP85-181RDB) by introduction of a hammerhead ribozyme Br J Cancer 1994 70: 239–243

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Kitazono M, Sumizawa T, Takebayashi Y et al. Multidrug resistance and the lung resistance–related protein in human colon carcinoma SW-620 cells J Natl Cancer Inst 1999 91: 1647–1653

    Article  CAS  PubMed  Google Scholar 

  16. Stuart DD, Kao GY, Allen TM . A novel, long-circulating, and functional liposomal formulation of antisense oligodeoxynucleotides targeted against MDR1 Cancer Gene Ther 2000 7: 466–475

    Article  CAS  PubMed  Google Scholar 

  17. Marthinet E, Divita G, Bernaud J, Rigal D, Baggetto LG . Modulation of the typical multidrug resistance phenotype by targeting the MED-1 region of human MDR1 promoter Gene Ther 2000 7: 1224–1233

    Article  CAS  PubMed  Google Scholar 

  18. Ruffner DE, Stormo GD, Uhlenbeck OC . Sequence requirements of the hammerhead RNA self-cleavage reaction Biochemistry 1990 29: 10695–10702

    Article  CAS  PubMed  Google Scholar 

  19. Kawabata S, Oka M, Shiozawa K et al. Breast cancer resistance protein directly confers SN-38 resistance of lung cancer cells Biochem Biophys Res Commun 2001 280: 1216–1223

    Article  CAS  PubMed  Google Scholar 

  20. Kowalski P, Wichert A, Holm PS, Dietel M, Lage H . Selection and characterization of a high activity ribozyme directed against the antineoplastic drug resistance–associated ABC-transporter BCRP/MXR/ABCG2 Cancer Gene Ther 2001 8: 185–192

    Article  CAS  PubMed  Google Scholar 

  21. Dietel M, Arps H, Lage H, Niendorf A . Membrane vesicle formation due to acquired mitoxantrone resistance in human gastric carcinoma cell line EPG85-257 Cancer Res 1990 50: 6100–6106

    CAS  PubMed  Google Scholar 

  22. Lage H, Helmbach H, Grottke C, Dietel M, Schadendorf D . DFNA5 (ICERE-1) contributes to acquired etoposide resistance in melanoma cells FEBS Lett 2001 494: 54–59

    Article  CAS  PubMed  Google Scholar 

  23. Stein U, Walther W, Laurencot CM, Scheffer GL, Scheper RJ, Shoemaker RH . Tumor necrosis factor-alpha and expression of the multidrug resistance–associated genes LRP and MRP J Natl Cancer Inst 1997 89: 807–813

    Article  CAS  PubMed  Google Scholar 

  24. Maliepaard M, Scheffer GL, Faneyte IF et al. Subcellular localization and distribution of the breast cancer resistance protein transporter in normal human tissues Cancer Res 2001 61: 3458–3464

    CAS  PubMed  Google Scholar 

  25. Scheffer GL, Maliepaard M, Pijnenborg AC et al. Breast cancer resistance protein is localized at the plasma membrane in mitoxantrone- and topotecan-resistant cell lines Cancer Res 2000 60: 2589–2593

    CAS  PubMed  Google Scholar 

  26. Skehan P, Storeng R, Scudiero D et al. New colorimetric cytotoxicity assay for anticancer-drug screening J Natl Cancer Inst 1990 82: 1107–1112

    Article  CAS  PubMed  Google Scholar 

  27. Allen JD, Brinkhuis RF, Wijnholds J, Schinkel AH . The mouse Bcrp1/Mxr/Abcp gene: amplification and overexpression in cell lines selected for resistance to topotecan, mitoxantrone, or doxorubicin Cancer Res 1999 59: 4237–4241

    CAS  PubMed  Google Scholar 

  28. Lage H, Perlitz C, Abele R et al. Enhanced expression of human ABC-transporter TAP is associated with cellular resistance to mitoxantrone FEBS Lett 2001 503: 179–184

    Article  CAS  PubMed  Google Scholar 

  29. Saxena SK, Ackerman EJ . Ribozymes correctly cleave a model substrate and endogenous RNA in vivo J Biol Chem 1990 265: 17106–17109

    CAS  PubMed  Google Scholar 

  30. Thierry AR, Rahman A, Dritschilo A . Overcoming multidrug resistance in human tumor cells using free and liposomally encapsulated antisense oligodeoxynucleotides Biochem Biophys Res Commun 1993 190: 952–960

    Article  CAS  PubMed  Google Scholar 

  31. Wichert A, Holm PS, Dietel M, Lage H . Selection of a high activity ribozyme against the cytostatic drug resistance associated glypican-3 using an in vitro assay containing total tumor RNA Cancer Gene Ther 1999 6: 263–270

    Article  CAS  PubMed  Google Scholar 

  32. Materna V, Holm PS, Dietel M, Lage H . Kinetic characterization of ribozymes directed against the cisplatin resistance-associated ABC-transporter cMOAT/MRP2/ABCC3 Cancer Gene Ther 2001 8: 176–184

    Article  CAS  PubMed  Google Scholar 

  33. Kellner U, Hutchinson L, Seidel A et al. Decreased drug accumulation in a mitoxantrone-resistant gastric carcinoma cell line in the absence of P-glycoprotein Int J Cancer 1997 71: 817–824

    Article  CAS  PubMed  Google Scholar 

  34. Lage H, Dietel M . Involvement of the DNA mismatch repair system in antineoplastic dug resistance J Cancer Res Clin Oncol 1999 125: 156–165

    Article  CAS  PubMed  Google Scholar 

  35. Zhou S, Schuetz JD, Bunting KD et al. The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype Nat Med 2001 7: 1028–1034

    Article  CAS  PubMed  Google Scholar 

  36. Hertel KJ, Pardi A, Uhlenbeck OC et al. Numbering system for the hammerhead Nucleic Acids Res 1992 20: 3252

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported in part by grants of the “NOVARTIS Stiftung für therapeutische Forschung”, the “Deutsche Forschungsgemeinschaft” (LA 1039/2-1) and the “Deutsche Krebshilfe” (10-1313-La3). We are grateful to RJ Scheper (Free University Amsterdam, the Netherlands) for providing the BCRP-specific mAbs BXP-21 and BXP-34.

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Authors and Affiliations

  1. Institute of Pathology, Charité Campus Mitte, Humboldt University Berlin, Berlin, D-10117, Germany

    Petra Kowalski & Hermann Lage

  2. Max-Delbrück-Center for Molecular Medicine, Berlin, D-13092, Germany

    Ulrike Stein

  3. Department of Pathology, Free University Medical Center, Amsterdam, 1007, The Netherlands

    George L Scheffer

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  1. Petra Kowalski
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  2. Ulrike Stein
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  3. George L Scheffer
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  4. Hermann Lage
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Correspondence to Hermann Lage.

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Kowalski, P., Stein, U., Scheffer, G. et al. Modulation of the atypical multidrug-resistant phenotype by a hammerhead ribozyme directed against the ABC transporter BCRP/MXR/ABCG2. Cancer Gene Ther 9, 579–586 (2002). https://doi.org/10.1038/sj.cgt.7700471

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